PTPRC / Receptor-type tyrosine-protein phosphatase C · IHC design guide

Design Immunohistochemistry for PTPRC

Plan CD45 IHC on paraffin sections using the catalog antibody's 1:50–1:200 dilution range (datasheet A00555-1) and assess selective cytoplasmic staining in lymphoid tissue and immune cells (HPA tissue IHC). Interpret that observed pattern alongside CD45's membrane topology (UniProt).

Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use

Immunohistochemistry protocol sheet for PTPRC (IHC for PTPRC): expected localisation Cytoplasmic tissue staining (HPA tissue IHC); membrane expected (UniProt), antibody A00555-1, validated IHC image, and IHC protocol steps — Printable **PTPRC** IHC protocol sheet — expected localisation Cytoplasmic tissue staining (HPA tissue IHC); membrane expected (UniProt), antibody A00555-1, controls and protocol steps. Open the full PTPRC IHC guide →

Navigate to: Protocols Staining pattern Controls Antibodies

PTPRC Immunohistochemistry Experimental Design Guide

Expected localisation, validated protocols, controls and antibodies — the at-a-glance facts below, then the full design guide.

Must know before staining

	Expected localisation	Cytoplasmic tissue staining (HPA tissue IHC); membrane expected (UniProt)
	Staining pattern	Selective cytoplasmic signal in lymphoid tissue and immune cells (HPA tissue IHC)
	Antigen retrieval	Citrate pH 6.0 HIER, heat-mediated (datasheet A00555-1)
	Positive control ⓘ	Appendix+4 more · see all
	Negative control ⓘ	Adipose tissue+4 more · see all

Important caveats

Reasons your staining may differ from the expected pattern.

	Fixation	Keep fixation consistent across sections (standard IHC practice; not target-specific); Matched tissue-IHC evidence does not establish the fixation claim. Validate the specimen-specific method before use. (selected-SKU IHC image A00555-1)
	Caveat	Immune-cell infiltrates can dominate the tissue signal (HPA tissue IHC)
	Regulation	Expression regulation not specified (UniProt)
	Isoform / epitope	8 isoforms; map extracellular vs cytoplasmic epitopes (UniProt)

Section 1

Recommended PTPRC IHC & IF Protocols

The catalog antibody uses citrate pH 6.0 retrieval (datasheet A00555-1). One published salivary gland IHC protocol provides an alternative (PMC10545362).

Recommended immunohistochemistry (IHC-P) protocol parameters
Sample	Paraffin-embedded human liver cancer tissue; fixative not specified (datasheet A00555-1)
Fixation	Image fixative and duration unreported (datasheet A00555-1); verify before use.
Sectioning	4–5 µm sections on charged slides (standard)
Deparaffinisation	Xylene, graded ethanol series to water (standard)
Antigen retrieval	Heat retrieval: Citrate pH 6.0 (datasheet A00555-1); 20 min, 95–100 °C (standard)
Peroxidase block	3% H2O2, 10 min, room temperature (standard)
Blocking	10% normal serum of the secondary host, 30 min, room temperature (standard)
Primary antibody	Rabbit anti-PTPRC, 1:50-1:200 (datasheet A00555-1)
Primary incubation	Overnight at 4 °C (standard)
Detection	HRP-polymer secondary, DAB chromogen 5–10 min (standard)
Counterstain	Hematoxylin, blue, dehydrate and mount (standard)
Expected result	PTPRC-positive staining in germinal center cells of appendix (HPA tissue IHC: High). HPA tissue profile: Selective cytoplasmic expression in lymphoid tissue and immune cells. No signal in the no-primary control.

💡Decision noteStart with citrate pH 6.0 heat retrieval (datasheet A00555-1); consider EDTA pH 8.0 for salivary gland sections (PMC10545362 methods).

Section 2

What Is the Expected PTPRC Staining Pattern?

PTPRC (CD45) is a cell-membrane protein with an extracellular region and cytoplasmic phosphatase domains (UniProt P08575 topology). In paraffin-section IHC, expect staining in immune and lymphoid cells, including germinal center cells and bone-marrow hematopoietic cells (HPA tissue IHC: High). HPA describes its observed tissue pattern as selective cytoplasmic expression and rates the IHC evidence Enhanced, with medium consistency between staining and RNA data (HPA tissue IHC).

What am I looking at on my slide?
Distinct staining in germinal center cells of tonsil, lymph node, or appendix, with surrounding cells easier to distinguish.	This matches HPA's High staining in those annotated cell populations (HPA tissue IHC). Assess the stained cells and their location on the section; a dark region alone does not establish that the expected cells are positive (standard IHC practice).
Staining outlines immune-cell membranes, or appears partly cytoplasmic in the expected cells.	Membrane staining fits PTPRC topology (UniProt P08575 topology). HPA reports selective cytoplasmic staining in tissue IHC (HPA tissue IHC), so cytoplasmic signal should be judged alongside cell identity and controls rather than rejected solely for lacking a crisp membrane rim.
Predominantly nuclear staining in paraffin-section IHC, without convincing staining of expected immune cells.	A nuclear-dominant IHC pattern does not match UniProt's membrane assignment or HPA's tissue-IHC profile (UniProt P08575 subcellular location; HPA tissue IHC). HPA's uncertain nucleoplasm finding comes from ICC-IF and does not validate a nuclear IHC interpretation (HPA subcellular ICC-IF).
Strong staining in adipocytes or respiratory epithelial cells, especially when expected immune cells show little signal.	HPA lists adipocytes in adipose tissue and respiratory epithelial cells in bronchus as Not detected (HPA tissue IHC). Consider nonspecific antibody binding or endogenous detection activity; review controls before interpreting those cells as PTPRC-positive (standard IHC practice).
Diffuse color across tissue, or no signal in expected cells of a known-positive section.	Diffuse color obscures cell-specific interpretation (standard IHC practice). Absence in a section containing HPA-positive germinal center or bone-marrow hematopoietic cells calls for a control and workflow check before concluding that the specimen lacks PTPRC (HPA tissue IHC; standard IHC practice).

💡Expected PTPRC appearanceCall a result positive when identifiable germinal center or hematopoietic cells show clear, cell-associated staining against a controlled background; these cells are High in HPA tissue IHC, while membrane localisation is assigned by UniProt (HPA tissue IHC; UniProt P08575 topology). Diffuse color or dominant staining of HPA Not detected cell populations is a suspect positive (HPA tissue IHC; standard IHC practice).

How each factor affects the staining
Compartment and epitope	PTPRC spans the membrane at residues 578–598, with extracellular residues 26–577 and cytoplasmic residues 599–1306 (UniProt P08575 topology). The supplied evidence does not identify the catalog antibody's epitope; do not infer whether it recognizes an extracellular or cytoplasmic region.
Observed IHC pattern and confidence	HPA calls tissue-IHC expression selective cytoplasmic and rates reliability Enhanced, while reporting medium consistency with RNA and staining in some unannotated cells or structures (HPA tissue IHC). Interpret localisation and unexpected cells with those limits in view.
Isoform coverage	UniProt lists 8 isoforms; isoforms 1, 2, 3, 5, and 7 are detected in thymocytes, whereas 4, 6, and 8 are not (UniProt P08575 isoforms and tissue specificity). Antibody recognition across isoforms is not established by this record.
Extracellular glycosylation	UniProt lists 17 glycosylation sites in the extracellular region (UniProt P08575 topology and glycosylation). Their presence alone does not establish epitope accessibility, retrieval requirements, or fixation sensitivity for this antibody.
IF/ICC Q&A: Should nuclear or vesicular IF signal define the expected IHC result?	No. HPA labels nucleoplasm and vesicle localisation uncertain in ICC-IF, and the listed antibody HPA000440 has ICC status Uncertain (HPA subcellular ICC-IF; HPA antibodies). Use the tissue-IHC evidence to interpret this paraffin-section guide (HPA tissue IHC).

Why is my staining missing, weak or wrong?
Situation	Likely cause	Next action
No staining in tonsil or lymph-node germinal center cells.	Those cells are High in HPA tissue IHC, so a negative result may reflect a failed IHC run or an absent suitable positive population (HPA tissue IHC; standard IHC practice).	Confirm the expected cells are present; compare an established positive section and reagent controls, then review retrieval and detection steps (standard IHC practice).
Broad brown signal covers the section and obscures individual cells.	Diffuse background can arise from nonspecific binding, insufficient washing, or endogenous detection activity (standard chromogenic IHC practice).	Inspect the negative control, blocking and wash steps, and the detection system's endogenous-activity control (standard chromogenic IHC practice).
Adipocytes or respiratory epithelial cells stain strongly.	These annotated cell types are Not detected in HPA tissue IHC; the signal may be nonspecific or due to endogenous detection activity (HPA tissue IHC; standard IHC practice).	Compare expected immune cells with the negative control and reassess antibody dilution and detection background (HPA tissue IHC; standard IHC practice).
Only nuclei appear strongly positive.	This is discordant with the UniProt membrane assignment and HPA's selective cytoplasmic tissue-IHC profile (UniProt P08575 subcellular location; HPA tissue IHC).	Check the counterstain and negative control, then verify that any proposed positive signal is cell-associated in expected immune cells (standard IHC practice; HPA tissue IHC).
Signal is faint in expected cells but background remains low.	Weak signal alone cannot distinguish limited assay sensitivity from lower staining in this particular section; HPA's High calls describe specified annotated populations, not every section (HPA tissue IHC; standard IHC practice).	Compare a positive control processed in the same run; review antibody dilution, antigen retrieval, incubation, and detection according to the antibody's IHC instructions (standard IHC practice).
Membrane rims are hard to see, but expected cells show cytoplasmic color.	HPA reports selective cytoplasmic tissue-IHC staining despite UniProt's membrane assignment (HPA tissue IHC; UniProt P08575 subcellular location).	Score cell identity and controlled contrast first; record the observed compartment without claiming that cytoplasmic staining proves a cytosolic PTPRC pool (HPA tissue IHC; standard IHC practice).

Source: UniProt P08575 Human Protein Atlas tissue Human Protein Atlas subcellular

Experiment evidence

IHC & IF images showing PTPRC staining

Immunohistochemistry of paraffin-embedded human liver cancer using CD45 Rabbit pAb at dilution of 1:50 .Perform high pressure antigen retrieval with 10 mM citrate buffer pH 6.0 before commencing with IHC staining protocol.

A00555-1 · IHCImmunohistochemistry of paraffin-embedded human liver cancer using CD45 Rabbit pAb at dilution of 1:50 .Perform high pressure antigen retrie

A00555-1 · IFImmunofluorescence analysis of Jurkat cells using CD45 Rabbit pAb at dilution of 1:100 . Blue: DAPI for nuclear staining.

IHC analysis of CD45 using anti-CD45 antibody (M00555-5). CD45 was detected in a paraffin-embedded section of human lung cancer tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH 8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 2 μg/ml mouse anti-CD45 Antibody (M00555-5) overnight at 4°C. Peroxidase Conjugated Goat Anti-mouse IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using HRP Conjugated Mouse IgG Super Vision Assay Kit (Catalog # SV0001) with DAB as the chromogen.

M00555-5 · IHCIHC analysis of CD45 using anti-CD45 antibody (M00555-5). CD45 was detected in a paraffin-embedded section of human lung cancer tissue. Heat

IF analysis of CD45 using anti-CD45 antibody (M00555-5). CD45 was detected in a paraffin-embedded section of human tonsli tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH 8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 5 μg/mL mouse anti-CD45 Antibody (M00555-5) overnight at 4°C. Biotin conjugated goat anti-mouse IgG (BA1001) was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using DyLight®488 Conjugated Avidin (BA1128). The section was counterstained with DAPI. Visualize using a fluorescence microscope and filter sets appropriate for the label used.

M00555-5 · IFIF analysis of CD45 using anti-CD45 antibody (M00555-5). CD45 was detected in a paraffin-embedded section of human tonsli tissue. Heat media

IHC analysis of CD45 using anti-CD45 antibody (PB9096). CD45 was detected in a paraffin-embedded section of human tonsil tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH 8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1 μg/ml rabbit anti-CD45 Antibody (PB9096) overnight at 4°C. Biotinylated goat anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC) (Catalog # SA1022) with DAB as the chromogen.

PB9096 · IHCIHC analysis of CD45 using anti-CD45 antibody (PB9096). CD45 was detected in a paraffin-embedded section of human tonsil tissue. Heat mediat

IF analysis of CD45 using anti-CD45 antibody (PB9096) CD45 was detected in paraffin-embedded section of human tonsil tissues. Heat mediated antigen retrieval was performed in citrate buffer (pH6, epitope retrieval solution ) for 20 mins. The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1μg/mL rabbit anti-CD45 Antibody (PB9096) overnight at 4°C. DyLight®488 Conjugated Goat Anti-Rabbit IgG (BA1127) was used as secondary antibody at 1:100 dilution and incubated for 30 minutes at 37°C. The section was counterstained with DAPI. Visualize using a fluorescence microscope and filter sets appropriate for the label used.

PB9096 · IFIF analysis of CD45 using anti-CD45 antibody (PB9096) CD45 was detected in paraffin-embedded section of human tonsil tissues. Heat mediated

IHC analysis of CD45 using anti-CD45 antibody (PB9785). CD45 was detected in a paraffin-embedded section of human tonsil tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH 8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 2 μg/ml rabbit anti-CD45 Antibody (PB9785) overnight at 4°C. Peroxidase Conjugated Goat Anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using HRP Conjugated Rabbit IgG Super Vision Assay Kit (Catalog # SV0002) with DAB as the chromogen.

PB9785 · IHCIHC analysis of CD45 using anti-CD45 antibody (PB9785). CD45 was detected in a paraffin-embedded section of human tonsil tissue. Heat mediat

IF analysis of CD45 using anti-CD45 antibody (PB9785). CD45 was detected in a paraffin-embedded section of human tonsil tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH 8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 5 μg/mL rabbit anti-CD45 Antibody (PB9785) overnight at 4°C. DyLight®488 Conjugated Goat Anti-Rabbit IgG (BA1127) was used as secondary antibody at 1:500 dilution and incubated for 30 minutes at 37°C. The section was counterstained with DAPI. Visualize using a fluorescence microscope and filter sets appropriate for the label used.

PB9785 · IFIF analysis of CD45 using anti-CD45 antibody (PB9785). CD45 was detected in a paraffin-embedded section of human tonsil tissue. Heat mediate

Immunohistochemistry analysis of paraffin-embedded human colon carcinoma tissue, using CD45 Antibody. The picture on the right is blocked with the synthesized peptide.

A00555 · IHCImmunohistochemistry analysis of paraffin-embedded human colon carcinoma tissue, using CD45 Antibody. The picture on the right is blocked wi

Immunofluorescence analysis of A549. 1, primary Antibody (red) was diluted at 1:200 (4°C overnight). 2, Goat Anti Rabbit IgG (H&L) - Alexa Fluor 594 Secondary antibody was diluted at 1:1000 (room temperature, 50min).3, Picture B: DAPI (blue) 10min.

A00555 · IFImmunofluorescence analysis of A549. 1, primary Antibody (red) was diluted at 1:200 (4°C overnight). 2, Goat Anti Rabbit IgG (H&L) - Alexa F

M00555-10 · IHCHuman spleen was stained with anti-CD45 rabbit antibody

IHC result Immunohistochemical staining of formalin fixed and paraffin embedded human tonsil tissue sections using anti-CD45 rabbit monoclonal antibody (clone RM291) at a 1:200 dilution.

M00555-2 · IHCIHC result Immunohistochemical staining of formalin fixed and paraffin embedded human tonsil tissue sections using anti-CD45 rabbit monoclon

Immunohistochemistry staining of human small intestine (paraffin-embedded sections) with anti-CD45 (MEM-28), 10 µg/ml. Commercially tested by LifeSpan BioSciences.

M00555-3 · IHCImmunohistochemistry staining of human small intestine (paraffin-embedded sections) with anti-CD45 (MEM-28), 10 µg/ml. Commercially tested b

Immunocytochemistry staining of human peripheral blood mononuclear cell using anti-human CD45 (MEM-28, green). DNA visualized by DAPI (blue)

M00555-3 · IFImmunocytochemistry staining of human peripheral blood mononuclear cell using anti-human CD45 (MEM-28, green). DNA visualized by DAPI (blue)

PA1596 · IHCAnti-CD45 antibody, PA1596, IHC(P) IHC(P): Human Tonsil Tissue

PA1596 · IFAnti-CD45 antibody, PA1596, ICC ICC: JURKAT Cell

IF analysis of CD45/PTPRC using anti-CD45/PTPRC antibody (A00555-3). CD45/PTPRC was detected in a paraffin-embedded section of human tonsil tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH 8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 4 μg/mL rabbit anti-CD45/PTPRC Antibody (A00555-3) overnight at 4°C. Biotin conjugated goat anti-rabbit IgG (BA1003) was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using DyLight®488 Conjugated Avidin (BA1128). The section was counterstained with DAPI. Visualize using a fluorescence microscope and filter sets appropriate for the label used.

A00555-3 · IFIF analysis of CD45/PTPRC using anti-CD45/PTPRC antibody (A00555-3). CD45/PTPRC was detected in a paraffin-embedded section of human tonsil

Immunofluorescence analysis of HeLa cells, using CD45 (Phospho-Ser1007) Antibody. The picture on the right is blocked with the phospho peptide.

A00555S1007 · IFImmunofluorescence analysis of HeLa cells, using CD45 (Phospho-Ser1007) Antibody. The picture on the right is blocked with the phospho pepti

IF analysis of CD45 using anti-CD45 antibody (M00555-6). CD45 was detected in a paraffin-embedded section of human tonsli tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH 8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 5 μg/mL mouse anti-CD45 Antibody (M00555-6) overnight at 4°C. Biotin conjugated goat anti-mouse IgG (BA1001) was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using DyLight®488 Conjugated Avidin (BA1128). The section was counterstained with DAPI. Visualize using a fluorescence microscope and filter sets appropriate for the label used.

M00555-6 · IFIF analysis of CD45 using anti-CD45 antibody (M00555-6). CD45 was detected in a paraffin-embedded section of human tonsli tissue. Heat media

IF analysis of CD45 using anti-CD45 antibody (PB9096-Dyl488) CD45 was detected in paraffin-embedded section of human tonsil tissues. Heat mediated antigen retrieval was performed in citrate buffer (pH6, epitope retrieval solution ) for 20 mins. The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1μg/mL rabbit anti-CD45 Antibody (PB9096-Dyl488) overnight at 4°C. DyLight488 Conjugated Goat Anti-Rabbit IgG (BA1127) was used as secondary antibody at 1:100 dilution and incubated for 30 minutes at 37°C. The section was counterstained with DAPI. Visualize using a fluorescence microscope and filter sets appropriate for the label used.

PB9096-Dyl488 · IFIF analysis of CD45 using anti-CD45 antibody (PB9096-Dyl488) CD45 was detected in paraffin-embedded section of human tonsil tissues. Heat me

HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000081237-PTPRC/tissue/cerebral+cortex#imid_19181243

HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000081237-PTPRC/tissue/colon#imid_19181177

HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000081237-PTPRC/tissue/liver#imid_19181170

HPA · KidneyHuman Protein Atlas IHC, Kidney. Source: https://www.proteinatlas.org/ENSG00000081237-PTPRC/tissue/kidney#imid_19181179

HPA · ICC-IF A-431Human Protein Atlas immunofluorescence, A-431, antibody HPA000440; green = PTPRC, blue = nucleus, red = microtubules.

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Real IHC / IF data behind the staining-pattern expectations in this section.

Sample controls for PTPRC IHC & IF

🧪Run lymph node first: germinal center cells should stain (HPA: High in lymph node germinal center cells). Use adipose tissue as the negative comparator and score its adipocytes, which should be unstained (HPA: Not detected in adipocytes); nonleukocyte stromal cells on the lymph node slide should also remain unstained (UniProt P08575: leukocyte common antigen).

Positive control tissue: Appendix (Germinal center cells, HPA High)

Negative control tissue: Adipose tissue (HPA Not detected)

ICC-IF cell lines (HPA subcellular resource): HPA ICC-IF images show PTPRC in A-431, HEL, U2OS, with annotated localisation: Nucleoplasm (uncertain), Vesicles (uncertain) (HPA subcellular).

Technical controls: Include a no-primary, secondary-only control, a concentration-matched rabbit IgG control (caption: rabbit polyclonal antibody), and a PTPRC knockout sample if available as a biological negative. For chromogenic lymph node IHC, quench endogenous peroxidase and distinguish background in densely packed germinal centers from cell-associated staining (HPA: High in lymph node germinal center cells).

⚠️Feasibility: No target-specific fixation window or fixation effect is reported, and the selected A00555-1 paraffin-section caption does not state its fixative (caption: fixative unreported). That caption specifies high-pressure retrieval in 10 mM citrate, pH 6.0, before staining at 1:50 (caption: A00555-1); it does not establish that retrieval is required under other conditions. The supplied evidence does not establish frozen sections or IF as easier; in lymph node, densely packed positive germinal center cells can complicate cell-level scoring (HPA: High in lymph node germinal center cells).

HPA tissue IHC evidence for PTPRC

Comprehensive Human Protein Atlas IHC scoring per tissue (reliability: Enhanced — Medium consistency between antibody staining and RNA expression data. Antibody staining in cells/structures not annotated, view images.). Rows are taken directly from the HPA tissue chart — click any row's HPA link to view the source.

Positive expression · recommended positive controls

Tissue	Cell type	Level	Evidence	Source
Appendix	Germinal center cells	High	Protein (IHC)	HPA →
Bone marrow	Hematopoietic cells	High	Protein (IHC)	HPA →
Lymph node	Germinal center cells	High	Protein (IHC)	HPA →
Skin	Fibrohistiocytic cells	High	Protein (IHC)	HPA →
Spleen	Cells in red pulp	High	Protein (IHC)	HPA →

Undetected expression · recommended negative controls

Tissue	Cell type	Level	Evidence	Source
Adipose tissue	Adipocytes	Not detected	Protein (IHC)	HPA →
Adrenal gland	Glandular cells	Not detected	Protein (IHC)	HPA →
Breast	Adipocytes	Not detected	Protein (IHC)	HPA →
Bronchus	Respiratory epithelial cells	Not detected	Protein (IHC)	HPA →
Caudate	Glial cells	Not detected	Protein (IHC)	HPA →

Section 3

Advanced PTPRC IHC Tips

Troubleshoot PTPRC/CD45 staining in paraffin section chromogenic IHC using the catalog image, tissue expression profile, and membrane topology as reference points.

Which retrieval condition should I try first when CD45 staining is weak?

Start with heat-mediated retrieval in 10 mM citrate, pH 6.0, for paraffin sections (datasheet A00555-1). Use high pressure as the initial heating method because the selected tissue image used it (caption A00555-1). Keep section thickness, detection chemistry, and antibody dilution constant while comparing retrieval duration on adjacent sections (standard IHC practice). The image's 1:50 dilution provides a documented starting point for that comparison (caption A00555-1). If staining remains weak, test longer heating or another buffer on paired sections, accepting a change only when immune-cell signal improves without tissue damage (HPA: immune-cell profile; standard IHC practice).

Could fixation explain absent CD45 staining in my paraffin sections?

Target-specific fixation sensitivity is unknown from the supplied evidence; the selected paraffin-section caption does not state a fixative (caption A00555-1). Record the fixative, fixation duration, processing history, and section age for each specimen before comparing results (standard IHC practice). Run a well-preserved positive control alongside the problem section with identical 10 mM citrate, pH 6.0 retrieval and detection (datasheet A00555-1; standard IHC practice). If both fail, examine retrieval, antibody, and detection performance before assigning a specimen-specific cause (standard IHC practice). If only the specimen fails, compare matched material with documented processing; its fixation history alone cannot establish a CD45-specific mechanism (standard IHC practice).

How should I judge membrane, cytoplasmic, or nuclear CD45 staining?

Expect a substantial cell-boundary component because CD45 is a cell-membrane protein with a transmembrane segment at residues 578–598 (UniProt P08575 topology and subcellular location). The tissue atlas also describes selective cytoplasmic staining in lymphoid tissue and immune cells, so assess cell identity and pattern together (HPA: tissue IHC profile). Its uncertain nucleoplasm and vesicle assignments come from subcellular imaging and warrant caution when interpreting tissue IHC (HPA: subcellular summary). Compare suspicious regions with neighboring immune cells at the same detection setting and counterstain intensity (standard IHC practice). Isolated strong nuclear staining without a plausible immune-cell pattern merits review of controls and morphology before being called positive (UniProt P08575 location; HPA: tissue IHC profile; standard IHC practice).

Can an epitope or splice variant explain inconsistent CD45 staining?

First identify the catalog antibody's mapped epitope, if available; the supplied caption does not establish its epitope or isoform coverage (caption A00555-1). PTPRC has 8 reported isoforms, and their detection in thymocytes differs, so equal staining across cell populations is not guaranteed (UniProt P08575 isoforms and tissue specificity). Its extracellular region spans residues 26–577 and carries 17 listed glycosylation sites; the cytoplasmic region spans 599–1306 (UniProt P08575 topology and glycosylation). Compare staining after the documented citrate retrieval on matched positive sections before attributing differences to epitope access (datasheet A00555-1; standard IHC practice). Resolve persistent discordance with an antibody of independently documented epitope specificity and appropriate controls (standard IHC practice).

How should I investigate discordant CD45 immunofluorescence and chromogenic IHC?

Treat IF as a separate assay: the selected 1:50 dilution and citrate retrieval document paraffin-section chromogenic IHC, not IF performance (caption A00555-1). Multiplex CD45 with an independently validated marker of the expected immune-cell population and inspect each channel separately before interpreting overlap (HPA: immune-cell profile; standard IF practice). Choose a fluorophore in a relatively low-autofluorescence channel for the specimen, and include unstained and single-color controls (standard IF practice). Determine epitope location before permeabilisation: an extracellular epitope may need little detergent, whereas a cytoplasmic epitope requires membrane access (UniProt P08575 topology; standard IF practice). Interpret nuclear or vesicular IF cautiously because those atlas assignments are uncertain (HPA: subcellular summary).

What causes diffuse brown staining outside expected CD45-positive cells?

Use a no-primary control to assess detection-system background, and block endogenous peroxidase before chromogen development (standard chromogenic IHC practice). Examine whether brown signal follows section edges, folds, debris, or damaged areas rather than recognizable cells (standard IHC practice). Titrate the antibody around the documented 1:50 image condition and keep retrieval at 10 mM citrate, pH 6.0 during the first comparison (caption A00555-1; datasheet A00555-1; standard IHC practice). Compare with a positive lymphoid control and cells expected to lack staining, such as adipocytes in adipose tissue (HPA: high in tonsil germinal-center cells; HPA: adipocytes not detected). Treat diffuse DAB color across unrelated cells as background until controls and cellular morphology support specificity (HPA: selective immune-cell profile; standard IHC practice).

How should I quantify CD45 staining when immune cells are unevenly distributed? ⚠ ANSWER MARKED FOR VERIFICATION

Define viable regions before scoring and exclude folds, edges, necrosis, and noncellular deposits (standard IHC practice). Report CD45-positive cell density per mm² of viable tissue and the percentage of positive cells using an explicitly stated denominator (standard IHC practice). If using an H-score, record intensity categories 0–3 and their cell percentages, yielding a possible 0–300 score (standard IHC practice). Separate immune-rich areas from other compartments when sampling because tissue staining is selective for immune cells and lymphoid structures (HPA: tissue IHC profile). Use the same retrieval, detection, threshold, and region-selection rules across specimens, and report the number of scored regions (standard IHC practice).

When is a CD45-positive area likely to be artefact?

A credible signal follows recognizable immune cells and can show a membrane-associated pattern consistent with CD45 topology (HPA: tissue IHC profile; UniProt P08575 location). Cytoplasmic tissue staining can also occur in the atlas profile, so compartment alone cannot decide specificity (HPA: tissue IHC profile). Question staining restricted to section edges, necrotic areas, or morphologically unrelated cells, especially if the no-primary control is positive (standard IHC practice). Review nuclear-only signal cautiously because the atlas nucleoplasm assignment is uncertain, while the curated UniProt location emphasizes membrane sites (HPA: subcellular summary; UniProt P08575 location). Check endogenous peroxidase blocking and compare staining with a positive immune-cell control before assigning biological meaning to diffuse DAB (standard chromogenic IHC practice; HPA: immune-cell profile).

Boster reagents

Best PTPRC / Receptor-type tyrosine-protein phosphatase C IHC Antibodies

Anti-PTPRC antibodies have IHC images from human tissue, including paraffin sections, and IF images from cells and tissue (catalog IHC and IF captions). Catalog reactivity also includes mouse and rat (A00555-1 reactivity).

Real IHC data Immunohistochemistry of paraffin-embedded human liver cancer using CD45 Rabbit pAb at dilution of 1:50 .Perform high pressure antigen retrieval with 10 mM citrate buffer pH 6.0 before commencing with IHC staining protocol.

Anti-PTPRC Antibody

Cat # A00555-1

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Real IHC data

Anti-CD45 Antibody ® (monoclonal, 2H3)

Cat # M00555-5

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Real IHC data

Anti-CD45/PTPRC Antibody ®

Cat # PB9096

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Real IHC data

Anti-CD45/PTPRC Antibody ®

Cat # PB9785

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Real IHC data Immunohistochemistry analysis of paraffin-embedded human colon carcinoma tissue, using CD45 Antibody. The picture on the right is blocked with the synthesized peptide.

Anti-CD45 PTPRC Antibody

Cat # A00555

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Real IHC data Human spleen was stained with anti-CD45 rabbit antibody

Anti-CD45 Rabbit Monoclonal Antibody

Cat # M00555-10

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A00555-1 shows human liver cancer paraffin sections; M00555-5 shows human lung cancer paraffin sections; PB9096 and PB9785 each show human tonsil paraffin sections (respective IHC captions). A00555 shows human colon carcinoma paraffin sections, while M00555-10 shows human spleen staining without a reported preparation method (respective IHC captions).

Which to pick: For tissue IHC, PB9096 provides a documented human tonsil paraffin example with EDTA pH 8.0 retrieval and 1 μg/ml primary antibody; its caption does not report the fixative (PB9096 IHC caption). For IF/ICC, A00555-1 lists both applications and shows Jurkat cell IF at 1:100 (A00555-1 applications; A00555-1 IF caption). For work across species, A00555 lists human, mouse and rat reactivity, although its own IHC image documents only human colon carcinoma paraffin sections and does not report the fixative (A00555 reactivity; A00555 IHC caption).

Each figure is that product's own IHC / IF validation image from its datasheet.

References

UniProt Consortium. UniProt entry P08575 (PTPRC_HUMAN, Receptor-type tyrosine-protein phosphatase C).
Human Protein Atlas. PTPRC tissue IHC expression (reliability: Enhanced).
Human Protein Atlas. PTPRC subcellular location (ICC-IF): Localized to the nucleoplasm and vesicles..
Human Protein Atlas. PTPRC antibody validation summary (6 antibodies).
Identification of key genes in salivary gland in Sjögren's syndrome complicated with Hashimoto thyroiditis: Common pathogenesis and potential diagnostic markers. Medicine 2023 — PMC10545362.
PTPRC functions as a prognosis biomarker in the tumor microenvironment of cutaneous melanoma. Scientific reports 2023 — PMC10667527.
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