PPP2R1B / Serine/threonine-protein phosphatase 2A 65 kDa regulatory subunit A beta isoform · IHC design guide

Design Immunohistochemistry for PPP2R1B

Plan PPP2R1B chromogenic IHC in paraffin sections using the reported cytoplasmic tissue pattern (HPA tissue IHC). Compare appendix glandular cells with high staining against salivary gland glandular cells with no detected staining, and start the catalog antibody at 1:25 (HPA tissue IHC; datasheet M05756).

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

Immunohistochemistry protocol sheet for PPP2R1B (IHC for PPP2R1B): expected localisation Cytoplasmic staining in most tissues (HPA tissue IHC), antibody M05756, validated IHC image, and IHC protocol steps — Printable **PPP2R1B** IHC protocol sheet — expected localisation Cytoplasmic staining in most tissues (HPA tissue IHC), antibody M05756, controls and protocol steps. Open the full PPP2R1B IHC guide →

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PPP2R1B 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 staining in most tissues (HPA tissue IHC)
	Staining pattern	Cytoplasmic staining in cells across most tissues (HPA tissue IHC)
	Antigen retrieval	Citrate pH 6 HIER, heat-mediated (datasheet M05756)
	Positive control ⓘ	Appendix+4 more · see all
	Negative control ⓘ	Salivary gland+1 more · see all

Important caveats

Reasons your staining may differ from the expected pattern.

	Fixation	Use formaldehyde-fixed paraffin sections (selected-SKU IHC image M05756); keep fixation consistent (standard IHC practice; not target-specific)
	Caveat	Staining–RNA agreement is medium; verify controls (HPA tissue IHC)
	Regulation	Liver RNA is tissue enhanced (HPA tissue RNA)
	Isoform / epitope	5 isoforms; epitope coverage is unspecified (UniProt; datasheet M05756)

Section 1

Recommended PPP2R1B IHC & IF Protocols

The catalog antibody protocol uses citrate pH 6 heat retrieval (datasheet M05756). The published IHC protocols below report additional sample preparation and detection details (PMC10908207; PMC4558495).

Recommended immunohistochemistry (IHC-P) protocol parameters
Sample	Formaldehyde-fixed, paraffin-embedded tissue sections (datasheet M05756; sample unspecified)
Fixation	Image formalin-fixed; duration unreported (datasheet M05756); 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 (datasheet M05756); 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	Mouse monoclonal (clone 1496CT356.164.25.226) anti-PPP2R1B, 1:25 (datasheet M05756)
Primary incubation	1 hours at 37°C (datasheet M05756)
Detection	HRP-polymer secondary, DAB chromogen 5–10 min (standard)
Counterstain	Hematoxylin, blue, dehydrate and mount (standard)
Expected result	PPP2R1B-positive staining in glandular cells of appendix (HPA tissue IHC: High). HPA tissue profile: Cytoplasmic expression in most tissues. No signal in the no-primary control.

💡Decision noteStart with citrate pH 6 heat retrieval for the catalog antibody (datasheet M05756); the published tumor staining protocol reports retrieval without conditions (PMC4558495).

Section 2

What Is the Expected PPP2R1B Staining Pattern?

PPP2R1B should show predominantly cytoplasmic staining in most tissues, with high staining reported in appendix glandular cells, tonsil germinal center cells, and late spermatids in testis (HPA: tissue IHC). HPA rates its tissue IHC profile Approved, with medium agreement between staining and RNA data; external verification is pending (HPA: reliability). PPP2R1B has no transmembrane segment, while UniProt does not assign a subcellular location (UniProt P30154: topology and localization).

What am I looking at on my slide?
Cytoplasmic staining in appendix glandular cells, tonsil germinal center cells, or late spermatids.	This fits the reported high IHC signal in these cells (HPA: tissue IHC). Compare stained cells with the section’s morphology and a negative detection control (general IHC practice).
A predominantly nuclear, extracellular, or sharply membrane-only chromogenic pattern.	This does not match HPA’s broad cytoplasmic tissue IHC profile (HPA: tissue IHC). Membrane signal alone is not conclusive: HPA also reports plasma membrane localization by ICC-IF (HPA: subcellular). Check controls before assigning the signal to PPP2R1B (general IHC practice).
Strong staining in salivary glandular cells or smooth muscle cells.	HPA reports these cell populations as not detected (HPA: tissue IHC). Cross-reactivity or endogenous detection activity is possible; review morphology and detection controls (general IHC practice). A single discordant section does not establish the cause.
Diffuse chromogen across cells and surrounding tissue, with poor cellular boundaries.	This is difficult to score as the reported cytoplasmic pattern (HPA: tissue IHC). Incomplete blocking, insufficient washing, or excess antibody or detection reagent can raise background (general IHC practice); compare the negative detection control.
No signal in appendix glandular cells, tonsil germinal center cells, or late spermatids.	These are reported high-staining populations (HPA: tissue IHC). Check that the expected cells are present, then review antibody conditions, retrieval, and detection with a control section (general IHC practice). HPA’s Approved profile still awaits external verification (HPA: reliability).

💡Expected PPP2R1B appearanceA positive IHC result is predominantly cytoplasmic, with high signal in the specified appendix glandular cells, tonsil germinal center cells, or late spermatids (HPA: tissue IHC); widespread diffuse deposit without cell boundaries is suspect (general IHC practice).

How each factor affects the staining
Tissue and cell population	HPA reports high signal in three distinct cell populations, medium signal in several others, and no detection in salivary glandular or smooth muscle cells (HPA: tissue IHC). Select and score controls by cell type rather than by whole-tissue color alone (general IHC practice).
IHC versus IF/ICC localization	For IHC, expect the reported cytoplasmic tissue pattern (HPA: tissue IHC). For IF/ICC, is membrane staining plausible? Yes: plasma membrane and cytosol are both approved locations (HPA: subcellular). Interpret each application against its own evidence.
Antibody validation	Two listed rabbit polyclonal antibodies have Approved IHC status; one also has Approved ICC status (HPA: antibodies). The tissue profile has medium agreement with RNA data and awaits external verification (HPA: reliability). Treat an unexpected pattern as unresolved until controls and independent evidence agree.
Molecular context	UniProt lists five isoforms, no signal peptide, no transmembrane segment, and no annotated subcellular location (UniProt P30154). These facts do not specify an IHC epitope, retrieval requirement, isoform-specific staining pattern, or fixation sensitivity.

Why is my staining missing, weak or wrong?
Situation	Likely cause	Next action
High-staining reference cells are blank.	Expected cells may be absent from the section, or an IHC workflow step may have failed (HPA: high-staining cells; general IHC practice).	Confirm cell identity on the counterstained section; check the antibody conditions, retrieval, and detection against a known-positive control (general IHC practice).
Most of the section is diffusely colored.	Nonspecific reagent binding, inadequate washing, or excess detection signal can obscure cell boundaries (general IHC practice).	Compare a negative detection control; review blocking, washes, antibody concentration, and chromogen development (general IHC practice).
Salivary glandular or smooth muscle cells stain strongly.	Those cells were not detected in the HPA tissue profile; staining may be nonspecific or arise from endogenous detection activity (HPA: tissue IHC; general IHC practice).	Check the cell identity and negative detection control; if using enzyme detection, assess the relevant endogenous activity (general IHC practice).
The signal appears predominantly nuclear.	A nuclear-dominant result differs from HPA’s cytoplasmic tissue profile; its source cannot be determined from localization alone (HPA: tissue IHC).	Inspect morphology, background, and controls; repeat with adjusted antibody and detection conditions if needed (general IHC practice).
Membrane outlining dominates the IHC slide.	HPA reports plasma membrane localization in ICC-IF, but describes tissue IHC as cytoplasmic in most tissues (HPA: subcellular; HPA: tissue IHC).	Record both compartment and cell type; compare IHC controls before deciding whether the membrane signal is specific (general IHC practice).
Positive and negative tissue controls look alike.	The run cannot resolve the HPA-reported difference between high-staining cells and cells listed as not detected (HPA: tissue IHC).	Verify that the intended cell populations are represented and assess negative detection controls, staining conditions, and scoring thresholds (general IHC practice).

Source: UniProt P30154 Human Protein Atlas tissue Human Protein Atlas subcellular

Experiment evidence

IHC & IF images showing PPP2R1B staining

M05756 staining PPP2R1B in human spleen sections by Immunohistochemistry (IHC-P -paraformaldehyde-fixed, paraffin-embedded sections). Tissue was fixed with formaldehyde and blocked with 3% BSA for 0. 5 hour at room temperature; antigen retrieval was by heat mediation with a citrate buffer (pH6). Samples were incubated with primary antibody (1/25) for 1 hours at 37°C. A undiluted biotinylated goat polyvalent antibody was used as the secondary antibody.

M05756 · IHCM05756 staining PPP2R1B in human spleen sections by Immunohistochemistry (IHC-P -paraformaldehyde-fixed, paraffin-embedded sections). Tissue

Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized U-2 OS (human bone osteosarcoma cell line) cells labeling PPP2R1B with M05756 at 1/25 dilution, followed by Dylight® 488-conjugated goat anti-mouse IgG secondary antibody at 1/200 dilution (green). Immunofluorescence image showing cytoplasm staining on U-2 OS cell line. Cytoplasmic actin is detected with Dylight® 554 Phalloidin at 1/100 dilution (red).The nuclear counter stain is DAPI (blue).

M05756 · IFImmunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized U-2 OS (human bone osteosarcoma cell line) cells la

HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000137713-PPP2R1B/tissue/cerebral+cortex#imid_5035612

HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000137713-PPP2R1B/tissue/colon#imid_5035538

HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000137713-PPP2R1B/tissue/liver#imid_5035523

HPA · PancreasHuman Protein Atlas IHC, Pancreas. Source: https://www.proteinatlas.org/ENSG00000137713-PPP2R1B/tissue/pancreas#imid_5035490

HPA · ICC-IF A-431Human Protein Atlas immunofluorescence, A-431, antibody HPA018908; green = PPP2R1B, blue = nucleus, red = microtubules.

← swipe / scroll for more images →

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

Sample controls for PPP2R1B IHC & IF

🧪Run appendix first and look for staining in glandular cells (HPA: High in appendix glandular cells). Use salivary gland as a negative tissue (HPA: Not detected in salivary gland glandular cells); on the appendix slide, neighboring cells without specific staining provide a visual background reference, not a confirmed target-negative population.

Positive control tissue: Appendix (Glandular cells, HPA High)

Negative control tissue: Salivary gland (HPA Not detected)

ICC-IF cell lines (HPA subcellular resource): HPA ICC-IF images show PPP2R1B in A-431, U-251MG, U2OS, RPTEC/TERT1, hTERT-RPE1 (serum starved), Sperm, with annotated localisation: Plasma membrane (approved), Cytosol (approved) (HPA subcellular).

Technical controls: Include a no-primary (secondary-only) control, a host- and clonality-matched isotype control, and PPP2R1B-knockout tissue or cells as a biological negative. Check endogenous peroxidase activity in appendix; if using the caption’s biotinylated secondary, also control for endogenous biotin (selected-SKU tissue-IHC caption: biotinylated secondary).

⚠️Feasibility: A target-specific fixation window or fixation effect is unreported, and the exact selected-SKU tissue-IHC caption leaves the fixative unreported (selected-SKU tissue-IHC caption: fixative not stated). Heat-mediated citrate retrieval at pH 6 was used in that paraffin-section example, but its necessity for PPP2R1B is unreported (selected-SKU tissue-IHC caption: heat-mediated citrate retrieval). There is no supplied comparison showing that frozen sections or IF are easier; for appendix IHC, check inflammatory-cell peroxidase background while assessing glandular-cell staining (HPA: High in appendix glandular cells).

HPA tissue IHC evidence for PPP2R1B

Comprehensive Human Protein Atlas IHC scoring per tissue (reliability: Approved — Medium consistency between antibody staining and RNA expression data. Pending external verification.). 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	Glandular cells	High	Protein (IHC)	HPA →
Testis	Elongated or late spermatids	High	Protein (IHC)	HPA →
Tonsil	Germinal center cells	High	Protein (IHC)	HPA →
Adrenal gland	Glandular cells	Medium	Protein (IHC)	HPA →
Bone marrow	Hematopoietic cells	Medium	Protein (IHC)	HPA →

Undetected expression · recommended negative controls

Tissue	Cell type	Level	Evidence	Source
Salivary gland	Glandular cells	Not detected	Protein (IHC)	HPA →
Smooth muscle	Smooth muscle cells	Not detected	Protein (IHC)	HPA →

Section 3

Advanced PPP2R1B IHC Tips

Troubleshoot PPP2R1B staining in paraffin sections by checking retrieval, cellular localisation and controls before comparing staining across samples.

What retrieval should I try when PPP2R1B staining is weak?

Start with heat-mediated retrieval in citrate buffer at pH 6 for paraffin sections (datasheet M05756). The selected tissue image used citrate retrieval, followed by 3% BSA blocking for 0.5 hour and primary antibody at 1:25 for 1 hour at 37°C (caption M05756). If staining remains weak, vary heating time on adjacent sections while keeping antibody concentration and detection conditions constant (standard IHC practice). Include a no-primary control and compare staining in glandular cells with the reported appendix pattern (standard IHC practice; HPA: High in appendix glandular cells). Excess heating can damage morphology, so judge signal alongside tissue preservation (standard IHC practice).

Could fixation explain faint or uneven PPP2R1B staining?

PPP2R1B-specific sensitivity to fixation is unknown from the supplied evidence (supplied fixation evidence). Record fixative, fixation duration and section processing for each specimen before comparing chromogenic intensity (standard IHC practice). If staining varies, process adjacent sections together and compare them using citrate retrieval at pH 6 with the same primary incubation and detection steps (datasheet M05756; standard IHC practice). Check morphology and staining at both the section edge and center, since uneven processing can complicate interpretation (standard IHC practice). Treat any apparent fixation effect as a hypothesis until matched specimens establish it experimentally (standard IHC practice).

Should PPP2R1B stain cytoplasm, membrane, or nucleus in tissue?

Expect predominantly cytoplasmic staining across many tissues in chromogenic IHC (HPA: cytoplasmic expression in most tissues). Plasma membrane and cytosol are approved locations in subcellular images, whereas UniProt supplies no subcellular annotation (HPA: approved plasma membrane and cytosol; UniProt P30154). Compare suspected membrane staining with cytoplasmic signal in the same cells and with the expected cell types, such as appendix glandular cells (HPA: High in appendix glandular cells; standard IHC practice). PPP2R1B has no transmembrane segment, so a sharp membrane-only outline needs independent validation before assignment to this protein (UniProt P30154 topology; standard IHC practice). Review a no-primary section for a matching outline (standard IHC practice).

Can this antibody distinguish PPP2R1B isoforms after retrieval?

PPP2R1B has 5 listed isoforms, but the supplied antibody evidence does not map its epitope to an isoform-specific sequence (UniProt P30154 isoforms; supplied antibody evidence). Obtain the immunogen sequence and align it with all 5 isoforms before interpreting a staining difference as isoform-specific (UniProt P30154 isoforms; standard IHC practice). The annotated N-terminal modification is acetylalanine at residue 2; its effect on this antibody cannot be inferred without epitope mapping (UniProt P30154 modified residues; standard IHC practice). Keep citrate retrieval at pH 6 constant during comparisons, then test specificity with an appropriate orthogonal control (datasheet M05756; standard IHC practice).

How should I cross-check a PPP2R1B IHC pattern by multiplex IF?

Use IF as a separate validation experiment and pair PPP2R1B with a marker identifying the cell population scored in IHC, such as appendix glandular cells (HPA: High in appendix glandular cells; standard IF practice). Select a fluorophore channel with low tissue autofluorescence and inspect an unstained section in every channel before interpreting overlap (standard IF practice). Because plasma membrane and cytosol are approved locations, compare staining with and without permeabilisation when the antibody epitope's membrane-facing side is unknown (HPA: approved plasma membrane and cytosol; standard IF practice). Confirm that the second marker and PPP2R1B signals occupy the intended cells, then return to matched chromogenic sections for scoring (standard IF and IHC practice).

How can I reduce diffuse brown PPP2R1B background?

First compare the stained section with no-primary and detection-only controls to locate background from the detection system (standard IHC practice). For a peroxidase and DAB workflow, block endogenous peroxidase before primary incubation and assess the no-primary control for residual brown signal (standard IHC practice). The selected image used 3% BSA for 0.5 hour and primary antibody at 1:25; these are starting conditions for that image, not an established optimum across specimens (caption M05756). If controls are clean but staining is widespread, titrate primary concentration while preserving citrate retrieval at pH 6 and evaluate cell-level patterns (datasheet M05756; standard IHC practice).

How should I quantify PPP2R1B staining across paraffin sections? ⚠ ANSWER MARKED FOR VERIFICATION

Define the cell population and region before scoring, because reported PPP2R1B levels differ among tissue cell types (HPA: tissue IHC profile; standard IHC practice). Use an H-score from 0–300 to combine the percentage of cells at each intensity, or report percent positive cells with a fixed positivity threshold (standard IHC practice). For spatial comparisons, report positive-cell density per mm² of viable tissue and normalise counts to the annotated region area (standard IHC practice). Keep section thickness, citrate retrieval at pH 6, exposure to chromogen and image settings consistent across cases (datasheet M05756; standard IHC practice). Score cytoplasmic and membrane-associated signal separately when both are present (HPA: tissue and subcellular localisation; standard IHC practice).

Which PPP2R1B staining patterns suggest an artefact?

A credible tissue pattern should be cellular and compatible with the predominantly cytoplasmic IHC profile, while membrane-associated signal warrants comparison with the approved subcellular locations (HPA: tissue IHC and subcellular profiles; standard IHC practice). Strong staining confined to section edges, necrotic areas or a no-primary control suggests processing or detection artefact (standard IHC practice). Check whether the labelled cells match the region being interpreted: appendix glandular cells are reported High, whereas salivary gland glandular cells are Not detected (HPA: tissue IHC). Residual endogenous peroxidase can produce brown signal in a DAB assay, so review the detection-only control before assigning faint staining to PPP2R1B (standard IHC practice). HPA tissue staining has Approved reliability with medium RNA concordance and pending external verification, so use independent validation for consequential conclusions (HPA: tissue IHC reliability).

Boster reagents

Best PPP2R1B / Serine/threonine-protein phosphatase 2A 65 kDa regulatory subunit A beta isoform IHC Antibodies

M05756 has documented IHC staining in human spleen sections and IF staining in human U-2 OS cells; its catalog lists human, mouse and rat reactivity (M05756 IHC caption; M05756 IF caption; catalog reactivity).

Real IHC data

Anti-PPP2R1B Antibody

Cat # M05756

View product →

The rendered SKU, M05756, shows IHC staining in paraffin-embedded human spleen sections; its caption reports formaldehyde fixation and also describes the sections as paraformaldehyde-fixed (M05756 IHC caption). Its IF image shows cytoplasmic staining in U-2 OS cells, and its listed applications include IHC-P and IF (M05756 IF caption; catalog applications).

Which to pick: Choose M05756 for paraffin-section IHC because its own figure documents staining in human spleen sections (M05756 IHC caption). Choose M05756 for IF/ICC because its own IF figure documents staining in U-2 OS cells (M05756 IF caption). For cross-species planning, M05756 lists human, mouse and rat reactivity, although the supplied IHC and IF images show human samples only (catalog reactivity; M05756 IHC and IF captions).

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

References

UniProt Consortium. UniProt entry P30154 (2AAB_HUMAN, Serine/threonine-protein phosphatase 2A 65 kDa regulatory subunit A beta isoform).
Human Protein Atlas. PPP2R1B tissue IHC expression (reliability: Approved).
Human Protein Atlas. PPP2R1B subcellular location (ICC-IF): Localized to the plasma membrane and cytosol..
Human Protein Atlas. PPP2R1B antibody validation summary (2 antibodies).
PPP2R1B abolishes colorectal cancer liver metastasis and sensitizes Oxaliplatin by inhibiting MAPK/ERK signaling pathway. Cancer cell international 2024 — PMC10908207.
MicroRNA-587 antagonizes 5-FU-induced apoptosis and confers drug resistance by regulating PPP2R1B expression in colorectal cancer. Cell death & disease 2015 — PMC4558495.
In search for biomarkers and potential drug targets for uterine serous endometrial cancer. Journal of cancer research and clinical oncology 2021 — PMC8076151.
Possible predisposition for colorectal carcinogenesis due to altered gene expressions in normal appearing mucosa from patients with colorectal neoplasia. BMC cancer 2019 — PMC6599319.
PubMed PMID:9795170 — UniProt-cited evidence.
PubMed PMID:9765152 — UniProt-cited evidence.
PubMed PMID:11313745 — UniProt-cited evidence.