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PFN1 / Profilin-1 · IHC design guide
Design Immunohistochemistry for PFN1
Plan chromogenic PFN1 staining in paraffin sections around its widespread cytoplasmic pattern and enrichment in immune-cell subsets (HPA tissue IHC). Use the catalog antibody’s IHC conditions and score staining by cell population (datasheet PB9313; HPA tissue IHC).
Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
PFN1 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) | |
| Staining pattern | Cytoplasm across tissues; strongest in immune-cell subsets (HPA tissue IHC) | |
| Antigen retrieval | EDTA pH 8.0 HIER, heat-mediated (datasheet PB9313) | |
| Positive control | Lung+4 more · see all | |
| Negative control | None in HPA (detected in all 45 tissues); use no-primary + isotype controls |
Important caveats
Reasons your staining may differ from the expected pattern.
| Fixation | Keep section fixation consistent (standard IHC practice; not target-specific) | |
| Caveat | Immune-rich regions may dominate the signal (HPA tissue IHC) | |
| Regulation | No expression regulator specified (UniProt) | |
| Isoform / epitope | No annotated isoforms; mature chain spans residues 2–140 (UniProt) |
Section 1
Recommended PFN1 IHC & IF Protocols
The catalog antibody protocol uses heat-mediated EDTA retrieval at pH 8.0 (datasheet: PB9313). The published IHC protocols below report conditions for oral, neural, breast, and bladder tissues (PMC5422140; PMC3923463; PMC3834125; PMC5342587).
| Sample | Paraffin-embedded human lung cancer tissue; fixative not specified (datasheet PB9313) |
| Fixation | Image fixative and duration unreported (datasheet PB9313); verify before use. |
| Sectioning | 4–5 µm sections on charged slides (standard) |
| Deparaffinisation | Xylene, graded ethanol series to water (standard) |
| Antigen retrieval | Heat retrieval: EDTA pH 8.0 (datasheet PB9313); 20 min, 95–100 °C (standard) |
| Peroxidase block | 3% H2O2, 10 min, room temperature (standard) |
| Blocking | 10% goat serum (datasheet PB9313) |
| Primary antibody | Rabbit anti-PFN1, 0.5-1μg/ml (datasheet PB9313) |
| Primary incubation | Overnight at 4 °C (datasheet PB9313) |
| Detection | Streptavidin-biotin complex (SABC), DAB chromogen (datasheet PB9313) |
| Counterstain | Hematoxylin, blue, dehydrate and mount (standard) |
| Expected result | PFN1-positive staining in alveolar cells type II of lung (HPA tissue IHC: High). HPA tissue profile: Ubiquitous cytoplasmic expression, most abundant in subsets of immune cells. No signal in the no-primary control. |
💡Decision noteStart with heat-mediated EDTA retrieval at pH 8.0 for the catalog antibody (datasheet: PB9313). For breast sections, one published protocol used citrate at pH 6.0 (PMC3834125).
Section 2
What Is the Expected PFN1 Staining Pattern?
PFN1 is a cytoplasmic, cytoskeleton-associated protein with no transmembrane segment (UniProt P07737). Expect cytoplasmic staining across many cell types, with stronger staining in subsets of immune cells (HPA: tissue IHC). HPA rates its tissue IHC profile Enhanced, while reporting medium consistency with RNA expression and pending external verification (HPA: tissue IHC reliability).
| Cytoplasmic staining in lymph-node germinal center cells and spleen red-pulp cells. | This fits two reported High IHC patterns (HPA: lymph node and spleen). Assess the labeled cells as well as intensity: PFN1 is broadly expressed, so staining elsewhere in the section can also be genuine (HPA: ubiquitous cytoplasmic expression). |
| Predominantly nuclear, membrane-rim, or extracellular staining, with little cytoplasmic signal. | That distribution conflicts with cytoplasm and cytoskeleton localization (UniProt P07737) and approved cytosolic localization in ICC-IF (HPA: subcellular). Review the antibody and detection controls before calling it PFN1; an unexpected compartment alone does not identify the artefact (general IHC practice). |
| Strong staining confined to a cell group reported as Low, without the expected stronger cells staining. | For example, glial cells in caudate and pancreatic exocrine glandular cells are reported Low (HPA: tissue IHC). Check cell identification, cross-reactivity, and endogenous detection activity (general IHC practice). Low is not an absence claim, and the HPA pattern awaits external verification (HPA: reliability). |
| Uniform color over cells, stroma, and clear spaces, obscuring cell boundaries. | Treat this as diffuse background until controls establish a cell-associated pattern (general IHC practice). PFN1 is expected in the cytoplasm (UniProt P07737; HPA: tissue IHC); broad expression does not explain equally strong deposit outside identifiable cells. |
| No signal in a correctly identified positive cell population. | Alveolar type II cells and lymph-node germinal center cells are reported High (HPA: tissue IHC). If both the sample and a suitable positive control lack signal, investigate the staining workflow before interpreting the study tissue as PFN1-negative (general IHC practice). |
💡Expected PFN1 appearanceCall a result consistent with PFN1 when staining is chiefly cytoplasmic and strong in a reported High population such as lymph-node germinal center cells; isolated nuclear or extracellular deposit is suspect (HPA: tissue IHC; UniProt P07737; general IHC practice).
| Cell population and tissue choice | Use reported High populations to judge whether the assay can reveal PFN1, while allowing weaker cells in the same or another section (HPA: tissue IHC). HPA lists no negative tissue here, so a Low population is a contrast, not a negative control (HPA: tissue IHC). |
| Localization and topology | PFN1 is cytoplasmic and cytoskeleton-associated, with no transmembrane segment (UniProt P07737); HPA approves cytosol as its main ICC-IF location (HPA: subcellular). These records support compartment interpretation but do not map this antibody's epitope or establish a PFN1-specific retrieval condition. |
| IHC antibody evidence | CAB037134 and CAB037140 each have Enhanced IHC validation (HPA: antibodies). That supports comparison with the reported tissue pattern, while HPA still describes medium staining-to-RNA consistency and pending external verification (HPA: tissue IHC reliability). |
| Retrieval and detection | Use the catalog antibody's IHC-P instructions to select retrieval and detection settings (general IHC practice). The supplied UniProt and HPA records provide no PFN1-specific fixation sensitivity or retrieval comparison; a weak section cannot establish either mechanism. |
| IF/ICC interpretation? | Expect cytosolic localization in ICC-IF; HPA lists A-431 and U-251MG images (HPA: subcellular). CAB037140 has Uncertain ICC status, while CAB037134 has no ICC status listed (HPA: antibodies). Consult the separate IF/ICC guide for that application. |
| Situation | Likely cause | Next action |
|---|---|---|
| The putative positive tissue is weak throughout. | The selected cells may have a reported Low level, or the IHC run may be insensitive (HPA: tissue IHC; general IHC practice). | Confirm cell identity and include a reported High population, such as lung alveolar type II cells, in the next run (HPA: lung). Then review catalog IHC-P settings and detection controls (general IHC practice). |
| Nuclei dominate while cytoplasm is faint. | The observed distribution disagrees with PFN1 localization; its precise cause is unresolved by the supplied records (UniProt P07737; HPA: subcellular). | Compare a no-primary control and a reported positive population, then reassess cell boundaries and counterstain before assigning localization (general IHC practice; HPA: tissue IHC). |
| Signal appears mainly in a reported Low cell group. | Cell identification, nonspecific binding, or endogenous detection activity may be involved; Low does not mean absent (HPA: tissue IHC; general IHC practice). | Verify the cell group and compare a reported High group. Check the appropriate reagent-only controls and detection blocking steps for the chosen chromogen system (HPA: tissue IHC; general IHC practice). |
| Diffuse deposit masks cytoplasmic detail. | Background from the staining workflow can prevent compartment scoring; the pattern alone cannot locate the source (general IHC practice). | Inspect no-primary and detection-only controls, then review blocking, washes, antibody dilution, and detection time according to the catalog IHC-P workflow (general IHC practice). |
| A known-positive population and its run control are blank. | A shared assay or detection failure is plausible before a biological absence call (general IHC practice; HPA: reported High populations). | Confirm section integrity, primary-antibody application, catalog IHC-P retrieval settings, and detection reagents; rerun with a reported High population (general IHC practice; HPA: tissue IHC). |
| An ICC-IF image disagrees with the IHC slide. | ICC evidence and tissue IHC evidence have different validation statuses: CAB037140 is Uncertain for ICC but Enhanced for IHC (HPA: antibodies). | Judge this paraffin-section result against IHC tissue patterns and controls; use the separate IF/ICC guide to assess the fluorescence experiment (HPA: tissue IHC; general IHC practice). |
Experiment evidence
IHC & IF images showing PFN1 staining
PB9313 · IHCIHC analysis of Profilin 1 using anti-Profilin 1 antibody (PB9313). Profilin 1 was detected in paraffin-embedded section of human lung cance
PB9313 · IHCIHC analysis of Profilin 1 using anti-Profilin 1 antibody (PB9313). Profilin 1 was detected in paraffin-embedded section of human intestinal
PB9313 · IFIF analysis of Profilin 1 using anti-Profilin 1 antibody (PB9313). Profilin 1 was detected in immunocytochemical section of U20S cells. Enzy
M01480 · IFImmunofluorescent analysis using the Antibody at 1:50 dilution.
HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000108518-PFN1/tissue/cerebral+cortex#imid_16269707
HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000108518-PFN1/tissue/colon#imid_16269633
HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000108518-PFN1/tissue/liver#imid_16269634
HPA · KidneyHuman Protein Atlas IHC, Kidney. Source: https://www.proteinatlas.org/ENSG00000108518-PFN1/tissue/kidney#imid_16269635
HPA · ICC-IF A-431Human Protein Atlas immunofluorescence, A-431, antibody CAB037140; green = PFN1, blue = nucleus, red = microtubules.
HPA · ICC-IF A-431Human Protein Atlas immunofluorescence, A-431, antibody CAB037140; green = PFN1, blue = nucleus, red = microtubules.
← swipe / scroll for more images →
Real IHC / IF data behind the staining-pattern expectations in this section.
Sample controls for PFN1 IHC & IF
🧪Run lung first and score PFN1 staining in alveolar type II cells (HPA: High in alveolar cells type II). HPA detects PFN1 in all 45 scored tissues, so there is no supported negative tissue; no cell population on the lung slide can be designated a true internal negative, and no-primary and isotype controls must establish background (HPA: no negative tissue rows; detected in all 45 tissues).
Positive control tissue: Lung (Alveolar cells type II, HPA High)
Negative control tissue: None in HPA: PFN1 is detected in all 45 scored tissues. Use a no-primary (secondary-only) and an isotype control instead.
ICC-IF cell lines (HPA subcellular resource): HPA ICC-IF images show PFN1 in A-431, U-251MG, with annotated localisation: Cytosol (approved) (HPA subcellular).
Technical controls: Include a no-primary, secondary-only slide and an isotype IgG control matched to the rabbit primary antibody’s host and clonality; use PFN1 knockout material or a validated peptide-block control to assess target specificity (caption: rabbit primary; standard IHC practice). With the caption’s biotin-based DAB detection, check endogenous peroxidase and biotin background in lung tissue (caption: SABC and DAB; standard IHC practice).
⚠️Feasibility: A target-specific fixation window is unreported, and the selected PB9313 paraffin-section caption does not state the fixative (caption: fixative not stated). The caption uses EDTA retrieval at pH 8.0, but supplies no comparison establishing retrieval dependence or showing that frozen sections or IF are easier; HPA reports cytosolic ICC-IF localization in A-431 and U-251MG cells (caption: EDTA retrieval; HPA: cytosol, A-431 and U-251MG). In lung, pigmented alveolar macrophages can resemble DAB staining, so assess cell morphology alongside the no-primary slide (standard IHC practice).
HPA tissue IHC evidence for PFN1
Comprehensive Human Protein Atlas IHC scoring per tissue (reliability: Enhanced — 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 |
|---|---|---|---|---|
| Lung | Alveolar cells type II | High | Protein (IHC) | HPA → |
| Lymph node | Germinal center cells | High | Protein (IHC) | HPA → |
| Spleen | Cells in red pulp | High | Protein (IHC) | HPA → |
| Tonsil | Non-germinal center cells | High | Protein (IHC) | HPA → |
| Adrenal gland | Glandular cells | Medium | Protein (IHC) | HPA → |
Undetected expression · recommended negative controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| None in HPA: PFN1 is detected in all 45 scored tissues. Use a no-primary (secondary-only) and an isotype control instead. | ||||
Section 3
Advanced PFN1 IHC Tips
Troubleshooting PFN1 staining in paraffin sections with chromogenic IHC, with one Q&A on IF multiplexing.
What should I change when PFN1 staining is weak after antigen retrieval?
Start with heat-mediated retrieval in EDTA pH 8.0 for paraffin sections (datasheet PB9313). The selected PB9313 tissue image used that buffer before overnight primary incubation at 4°C, but its caption gives no retrieval duration, so optimise heating time on matched sections (datasheet PB9313; standard IHC practice). Include a section processed without primary antibody and a positive tissue control in each comparison; lung alveolar type II cells offer a reported high-expression reference (standard IHC practice; HPA: High in lung alveolar type II cells). If staining remains weak, compare a second retrieval condition as a fallback while keeping section thickness, detection and exposure to chromogen constant (standard IHC practice).
Could fixation explain weak or uneven PFN1 staining?
Target-specific sensitivity of PFN1 staining to fixation is unknown from the supplied evidence; the PB9313 paraffin-section caption does not state a fixative (datasheet PB9313). Record the actual fixative and fixation time for each specimen, then compare matched sections processed with the same EDTA pH 8.0 retrieval and detection conditions (standard IHC practice; datasheet PB9313). Check whether loss of staining tracks fixation batches, section edges or uneven reagent coverage before changing the primary antibody concentration (standard IHC practice). Do not infer a PFN1-specific fixation effect from its cytoplasmic location, modified residues or tissue-expression pattern (UniProt P07737: cytoplasm and modified residues; HPA: ubiquitous cytoplasmic expression).
Where should convincing PFN1 signal appear in a paraffin section?
Expect predominantly cytoplasmic staining: PFN1 is assigned to the cytoplasm and cytoskeleton, and tissue IHC reports ubiquitous cytoplasmic expression (UniProt P07737: subcellular location; HPA: tissue profile). Its approved cellular location is the cytosol, and the record has no transmembrane segment (HPA: Cytosol approved; UniProt P07737: topology). Evaluate staining within identified cells against the section’s morphology and counterstain, rather than treating diffuse extracellular colour as PFN1 localisation (standard IHC practice). If a sample shows chiefly nuclear, membranous or extracellular signal, repeat staining with a no-primary control and inspect adjacent tissue before assigning a biological explanation (standard IHC practice; UniProt P07737: subcellular location).
How should I assess an unexpected PFN1 staining pattern?
The supplied PFN1 record lists 0 isoforms and no annotated domain, so it does not support assigning a staining pattern to a named isoform (UniProt P07737: isoforms and domains). Its listed modified residues include acetylation at positions 2 and 105 and phosphorylation at 28, 57 and 85; the PB9313 caption does not map its epitope (UniProt P07737: modified residues; datasheet PB9313). Compare the pattern across retrieval conditions and, if available, an independently validated antibody with a known epitope (standard IHC practice). Do not attribute altered chromogenic intensity to any one modification without direct epitope or perturbation evidence (standard IHC practice).
How can I assess PFN1 localisation in a multiplex IF experiment?
For this secondary application, pair PFN1 with a marker identifying the cell population being evaluated; the IHC profile reports high signal in lung alveolar type II cells and lymph-node germinal-center cells (HPA: tissue IHC). Place the weaker marker in a brighter fluorophore channel and inspect unstained tissue to choose channels with less autofluorescence (standard IF practice). PFN1 is cytosolic and has no transmembrane segment, so use a permeabilisation condition that permits antibody access to intracellular epitopes, then check whether cell boundaries remain interpretable (HPA: Cytosol approved; UniProt P07737: topology; standard IF practice). Include single-stain controls to assess spectral bleed-through and a no-primary control to assess background (standard IF practice).
How do I reduce diffuse brown signal without losing PFN1 staining?
Separate tissue background from antibody-dependent staining with a no-primary control, and inspect regions away from section edges and folds (standard IHC practice). The selected PB9313 image used 10% goat serum blocking, 1 µg/mL primary overnight at 4°C, a biotinylated secondary and DAB development (datasheet PB9313). Because that workflow uses biotin-based detection, check a detection-only control for tissue-associated signal and assess endogenous peroxidase blocking as part of the general chromogenic workflow (datasheet PB9313; standard IHC practice). If background persists, titrate primary concentration and chromogen development on matched sections while retaining a reported positive tissue reference (standard IHC practice; HPA: High in lung alveolar type II cells).
What is a defensible way to quantify PFN1 IHC across samples? ⚠ ANSWER MARKED FOR VERIFICATION
Define the cell population and tissue area before scoring, then report the percentage of positive cells together with an intensity-based H-score or positive-cell density per mm² (standard IHC practice). Measure within comparable viable regions and normalise cell counts to the number of eligible cells or measured area, rather than to total image pixels (standard IHC practice). Keep retrieval, antibody incubation, chromogen development and image settings consistent across the comparison; the PB9313 example used EDTA pH 8.0 retrieval and 1 µg/mL primary (standard IHC practice; datasheet PB9313). Report cell-type composition alongside the score because PFN1 tissue staining is ubiquitous but especially abundant in subsets of immune cells (HPA: tissue profile).
When is apparent PFN1 positivity likely to be artefactual?
A credible PFN1 result should chiefly occupy cell cytoplasm and follow identifiable cell morphology, consistent with its cytoplasmic and cytosolic annotations (UniProt P07737: subcellular location; HPA: Cytosol approved). Check which cells are stained before comparing samples: tissue IHC describes ubiquitous expression with particularly abundant signal in subsets of immune cells (HPA: tissue profile). Treat staining confined to cut edges, folds or necrotic areas as suspect, and compare it with matched intact regions and a no-primary control (standard IHC practice). Persistent brown signal in a detection-only control warrants review of endogenous enzyme activity or the biotin-based detection workflow before assigning PFN1 positivity (standard IHC practice; datasheet PB9313).
Boster reagents
Best PFN1 / Profilin-1 IHC Antibodies
The catalog lists anti-PFN1 antibodies for IHC and IF/ICC in human, mouse and rat (catalog: PB9313 and M01480 applications and reactivity); tissue IHC images document human paraffin sections (PB9313 image captions).
Real IHC data
Anti-Profilin 1/PFN1 Antibody ®
Cat # PB9313
Real IF data
Anti-Profilin1 PFN1 Rabbit Monoclonal Antibody
Cat # M01480
PB9313 has IHC images from human lung and intestinal cancer paraffin sections and an IF image from U20S cells (PB9313 image captions). M01480 lists IHC and IF/ICC applications, with an IF image whose caption does not identify the sample (M01480 catalog applications and IF caption).
Which to pick: For tissue IHC, choose PB9313: its human paraffin-section captions document EDTA retrieval at pH 8.0 and staining at 1 μg/ml; the fixative is unreported (PB9313 IHC image captions). For IF/ICC, PB9313 has a documented cell image at 2 μg/ml, while M01480 is a monoclonal option with an IF image at 1:50 and no sample named in its caption (PB9313 IF caption; M01480 catalog and IF caption). Both list human, mouse and rat reactivity, but the supplied IHC images document human tissue only (PB9313 and M01480 catalog reactivity; PB9313 IHC image captions).
Each figure is that product's own IHC / IF validation image from its datasheet.
References
- UniProt Consortium. UniProt entry P07737 (PROF1_HUMAN, Profilin-1).
- Human Protein Atlas. PFN1 tissue IHC expression (reliability: Enhanced).
- Human Protein Atlas. PFN1 subcellular location (ICC-IF): Localized to the cytosol..
- Human Protein Atlas. PFN1 antibody validation summary (2 antibodies).
- A loss of profilin-1 in late-stage oral squamous cell carcinoma. Journal of oral pathology & medicine : official publication of the International Association of Oral Pathologists and the American Academy of Oral Pathology 2017 — PMC5422140.
- Profilin-1 mutations are rare in patients with amyotrophic lateral sclerosis and frontotemporal dementia. Amyotrophic lateral sclerosis & frontotemporal degeneration 2013 — PMC3923463.
- Profilin-1 downregulation has contrasting effects on early vs late steps of breast cancer metastasis. Oncogene 2014 — PMC3834125.
- Silencing of Profilin-1 suppresses cell adhesion and tumor growth via predicted alterations in integrin and Ca2+ signaling in T24M-based bladder cancer models. Oncotarget 2016 — PMC5342587.
- PubMed PMID:3356709 — UniProt-cited evidence.
- PubMed PMID:20736409 — UniProt-cited evidence.
- PubMed PMID:14702039 — UniProt-cited evidence.
Immunohistochemistry protocol sheet for PFN1 (Profilin-1). Expected localisation Cytoplasmic tissue staining (HPA tissue IHC). Antigen retrieval: EDTA pH 8.0 HIER, heat-mediated (datasheet PB9313). Validated IHC antibody: PB9313 at 0.5-1μg/ml (the product's own IHC validation image is shown). Controls (HPA): positive Lung (Alveolar cells type II); negative None in HPA (detected in all 45 tissues); use no-primary + isotype controls. IHC-P workflow on paraffin sections; fixative unspecified. Protocol steps: 1. Sample: Paraffin-embedded human lung cancer tissue; fixative not specified (datasheet PB9313); 2. Fixation: Image fixative and duration unreported (datasheet PB9313); verify before use.; 3. Sectioning: 4–5 µm sections on charged slides (standard); 4. Deparaffinisation: Xylene, graded ethanol series to water (standard); 5. Antigen retrieval: Heat retrieval: EDTA pH 8.0 (datasheet PB9313); 20 min, 95–100 °C (standard); 6. Peroxidase block: 3% H2O2, 10 min , room temperature (standard); 7. Blocking: 10% goat serum (datasheet PB9313); 8. Primary antibody: Rabbit anti-PFN1, 0.5-1μg/ml (datasheet PB9313); 9. Primary incubation: Overnight at 4 °C (datasheet PB9313). Watch for: Fixation: Keep section fixation consistent (standard IHC practice; not target-specific); Caveat: Immune-rich regions may dominate the signal (HPA tissue IHC); Isoform / epitope: No annotated isoforms; mature chain spans residues 2–140 (UniProt); Regulation: No expression regulator specified (UniProt). Evidence: UniProt P07737, HPA tissue IHC · Enhanced, HPA ICC-IF · cytosol., Datasheet PB9313, PMC5422140, PMC3923463. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/PFN1/immunohistochemistry-ihc-guide.