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ARPC5 / Actin-related protein 2/3 complex subunit 5 · IHC design guide
Design Immunohistochemistry for ARPC5
Plan ARPC5 chromogenic IHC in paraffin sections using the observed cytoplasmic tissue profile (HPA tissue IHC). Compare staining with appropriate controls, mindful that the tissue evidence cautions the antibody may detect products of more than one gene (HPA tissue IHC).
Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
ARPC5 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 observed (HPA tissue IHC); nuclear location annotated (UniProt) | |
| Staining pattern | Cytoplasmic staining across several tissue types (HPA tissue IHC) | |
| Antigen retrieval | Tris-EDTA pH 9.0 HIER, 95–98 °C, 20 min (rule: nuclear antigen) | |
| Positive control | Testis+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 paraffin-section fixation consistent (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 M02096) | |
| Caveat | Antibody staining may reflect more than one gene (HPA tissue IHC) | |
| Regulation | Expression regulation unannotated (UniProt) | |
| Isoform / epitope | 2 isoforms; epitope impact is unknown (UniProt) |
Section 1
Recommended ARPC5 IHC & IF Protocols
The catalog antibody’s IHC-P protocol is accompanied by published ARPC5 staining methods for liver cancer, rat frontal cortex, and glioma (PMC9485570; PMC3584917; PMC10548738).
| Sample | Paraffin-embedded human colon tissue; fixative not specified (datasheet M02096) |
| Fixation | Image fixative and duration unreported (datasheet M02096); verify before use. |
| Sectioning | 4–5 µm sections on charged slides (standard) |
| Deparaffinisation | Xylene, graded ethanol series to water (standard) |
| Antigen retrieval | Heat-induced epitope retrieval in Tris-EDTA buffer, pH 9.0, 20 min at 95–98 °C (standard rule: nuclear antigen) |
| 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 monoclonal (clone CCD-1) anti-ARPC5, 1:50 (datasheet M02096) |
| 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 | ARPC5-positive staining in elongated or late spermatids of testis (HPA tissue IHC: High). HPA tissue profile: General cytoplasmic expression in several different tissue types. No signal in the no-primary control. |
💡Decision noteStart with Tris-EDTA at pH 9.0, 95–98 °C for 20 min (page retrieval protocol). Citrate at pH 6.0 is a published alternative (PMC3584917).
Section 2
What Is the Expected ARPC5 Staining Pattern?
ARPC5 staining in paraffin sections should be predominantly cytoplasmic across several tissue types, with particularly strong staining reported in elongated or late spermatids (HPA: general cytoplasmic expression; High in testis spermatids). Cytoskeletal and nuclear localization are also annotated (UniProt O15511: subcellular location). ARPC5 has no transmembrane segment (UniProt O15511: topology). Interpret the pattern cautiously: HPA rates tissue IHC Approved but reports low agreement with RNA data and a possible multi-gene antibody target (HPA: reliability description).
| Strong cytoplasmic staining in elongated or late spermatids, with moderate staining in some glandular or neuronal cells. | This fits the reported cell-level pattern: spermatids are High; adrenal, appendix and breast glandular cells and selected neuronal cells are Medium (HPA: tissue IHC). Score the named cell population and compartment, since an entire tissue need not stain uniformly (HPA: cell-level tissue IHC). |
| Predominantly nuclear staining, or staining confined to a compartment without a credible cytoplasmic pattern. | Check morphology and controls before calling it ARPC5: the tissue IHC profile is generally cytoplasmic (HPA: tissue IHC). Nuclear ARPC5 is annotated, however, so nuclear signal alone cannot prove artefact (UniProt O15511: subcellular location). HPA IF also reports membrane, junction, vesicle and cytosol localization; those images do not establish an IHC pattern (HPA: subcellular ICC-IF). |
| Strong staining in adipocytes or skin fibroblasts while expected positive cells are weak. | These cell types were Not detected in the reported sections (HPA: adipose tissue adipocytes; skin fibroblasts). Consider nonspecific binding, endogenous chromogen activity or an antibody recognizing another target; this is a diagnostic possibility, not a demonstrated mechanism (HPA: tissue IHC reliability description). Recheck cell identification and controls before scoring. |
| Diffuse brown signal across tissue, including extracellular spaces or many unrelated cell types. | A broad haze obscures the cell-level, mainly cytoplasmic pattern reported for ARPC5 (HPA: tissue IHC profile). It can arise from detection background or insufficient blocking or washing (general IHC practice). Compare a primary-antibody omission control and inspect whether signal follows cellular boundaries before interpreting staining. |
| No signal in elongated or late spermatids in an otherwise interpretable testis section. | That conflicts with the High staining reported for these cells (HPA: testis tissue IHC). First confirm that the relevant cells are present and distinguish absent target staining from a failed detection run using appropriate run controls (general IHC practice). HPA's Approved rating and stated consistency limits mean a single negative slide is not definitive (HPA: reliability description). |
💡Expected ARPC5 appearanceCall a convincing positive result cell-associated, mainly cytoplasmic staining, strongest in elongated or late spermatids and potentially moderate in the listed glandular or neuronal cells; diffuse extracellular haze or strong signal in reported Not detected cell types is suspect (HPA: tissue IHC profile and cell-level staining).
| Tissue and cell choice | Testis elongated or late spermatids offer a reported High comparison; adipocytes and skin fibroblasts were Not detected (HPA: tissue IHC). Use the specified cells when judging a section, since a tissue label alone does not identify the scored population (HPA: cell-level tissue IHC). |
| Evidence strength and antibody identity | One listed antibody, HPA022013, is Approved for IHC; HPA031972 has no listed IHC status (HPA: antibody validation). HPA also warns of low staining-to-RNA consistency and a possible multi-gene target, so morphology and controls matter even when a slide resembles the reference (HPA: reliability description). |
| Compartment and isoforms | ARPC5 is annotated in cytoplasm, cytoskeleton, cell projections and nucleus, has no transmembrane segment, and has two isoforms (UniProt O15511: location, topology, isoforms). These annotations do not establish which isoform an IHC antibody detects or make every compartment equally expected in paraffin sections. |
| Epitope and retrieval | No epitope or ARPC5-specific antigen retrieval condition is supplied (UniProt O15511 record; HPA: supplied tissue IHC data). Choose retrieval from the IHC-validated antibody's own instructions, then assess tissue morphology and controls (general IHC practice); do not infer retrieval needs from topology or staining intensity. |
| IF/ICC Q&A: what localization should be expected? | HPA reports approved vesicle, plasma membrane, cell junction and cytosol localization in ICC-IF (HPA: subcellular ICC-IF). Those observations can inform interpretation of IF images, but they are a separate application and do not prescribe an IF protocol or override the generally cytoplasmic tissue IHC profile (HPA: tissue IHC). |
| Situation | Likely cause | Next action |
|---|---|---|
| The testis comparison lacks staining in elongated or late spermatids. | The expected cells may be absent from the section, or the IHC run may have failed; High spermatid staining is the reported reference (HPA: testis tissue IHC). | Verify spermatid morphology, inspect an appropriate run control, and check the IHC-validated antibody's retrieval, dilution and detection instructions (general IHC practice). Avoid assigning an ARPC5-specific fixation effect; none is supplied. |
| Brown staining is widespread, granular or extracellular. | Detection background or residual endogenous enzyme activity can mimic chromogenic staining (general IHC practice). The reference is general cellular cytoplasmic expression (HPA: tissue IHC profile). | Compare primary-antibody omission and detection controls; review blocking, washing and chromogen development (general IHC practice). Score only signal tied to identifiable cells and a credible compartment. |
| A reported Not detected cell population stains strongly. | Nonspecific detection or recognition of another protein is possible, particularly given HPA's multi-gene target warning (HPA: reliability description). The reported status is cell-specific, not a claim that the whole tissue is blank (HPA: tissue IHC). | Confirm the cell type, compare a reported positive population and negative control, and seek independent antibody or orthogonal support before assigning the signal to ARPC5 (general IHC practice). |
| Only nuclear staining is visible. | Nuclear localization is biologically annotated, but it does not match the general cytoplasmic tissue IHC profile by itself (UniProt O15511: location; HPA: tissue IHC). | Inspect nuclear counterstain alignment, cytoplasmic visibility and omission controls (general IHC practice). Record nuclear staining separately; seek supporting evidence before calling it specific ARPC5. |
| Staining differs from an ICC-IF image. | HPA ICC-IF reports vesicles, membrane, junctions and cytosol, whereas tissue IHC is summarized as generally cytoplasmic (HPA: subcellular ICC-IF; tissue IHC). The applications show different preparations and readouts (general IHC/IF practice). | Judge the paraffin section against cell-level tissue IHC references and its own controls (HPA: tissue IHC; general IHC practice). Treat the IF locations as context, not a required chromogenic pattern. |
Experiment evidence
IHC & IF images showing ARPC5 staining
M02096 · IHCImmunohistochemical analysis of paraffin-embedded human colon, using p16 ARC Antibody.
M02096-1 · IFImmunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized U-2 OS (human osteosarcoma cell line) cells labelin
HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000162704-ARPC5/tissue/cerebral+cortex#imid_5785972
HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000162704-ARPC5/tissue/colon#imid_5785890
HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000162704-ARPC5/tissue/liver#imid_5785899
HPA · KidneyHuman Protein Atlas IHC, Kidney. Source: https://www.proteinatlas.org/ENSG00000162704-ARPC5/tissue/kidney#imid_5785892
HPA · ICC-IF A-431Human Protein Atlas immunofluorescence, A-431, antibody HPA031972; green = ARPC5, blue = nucleus, red = microtubules.
HPA · ICC-IF A-431Human Protein Atlas immunofluorescence, A-431, antibody HPA031972; green = ARPC5, blue = nucleus, red = microtubules.
← swipe / scroll for more images →
Real IHC / IF data behind the staining-pattern expectations in this section.
Sample controls for ARPC5 IHC & IF
🧪Run testis first: elongated or late spermatids should stain (HPA: High). Use adipose tissue as the negative tissue, focusing on adipocytes (HPA: Not detected); on the testis slide, assess neighboring non-spermatid cells for counterstain with absent or background-level chromogen, without assuming they are confirmed ARPC5-negative cells (HPA: no negative testis cell population supplied).
Positive control tissue: Testis (Elongated or late spermatids, HPA High)
Negative control tissue: Adipose tissue (HPA Not detected)
ICC-IF cell lines (HPA subcellular resource): HPA ICC-IF images show ARPC5 in A-431, U-251MG, U2OS, with annotated localisation: Vesicles (approved), Plasma membrane (approved), Cell Junctions (approved), Cytosol (approved) (HPA subcellular).
Technical controls: Include a no-primary, secondary-only control and a control matched to the primary antibody’s host species and class: an isotype control for a monoclonal antibody or nonimmune IgG for a polyclonal antibody (standard IHC practice). Use ARPC5 knockout material, if available, as a biological specificity control (standard IHC practice).
Block endogenous peroxidase in testis sections for chromogenic detection; block endogenous biotin if using avidin–biotin detection (standard IHC practice).
⚠️Feasibility: A target-specific fixation window or fixation effect is unreported, and the selected M02096 paraffin-section colon caption does not state the fixative (caption: paraffin-embedded human colon; fixative unreported). ARPC5-specific antigen-retrieval dependence is unreported; assess retrieval empirically with the tissue controls (supplied evidence: no retrieval conditions). The evidence does not establish whether frozen sections or IF are easier; in testis, score the specified spermatids rather than averaging across all cells (HPA: High in elongated or late spermatids).
HPA tissue IHC evidence for ARPC5
Comprehensive Human Protein Atlas IHC scoring per tissue (reliability: Approved — Low consistency between antibody staining and RNA expression data. Caution, targets protein from more than one gene.). 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 |
|---|---|---|---|---|
| Testis | Elongated or late spermatids | High | Protein (IHC) | HPA → |
| Adrenal gland | Glandular cells | Medium | Protein (IHC) | HPA → |
| Appendix | Glandular cells | Medium | Protein (IHC) | HPA → |
| Breast | Glandular cells | Medium | Protein (IHC) | HPA → |
| Bronchus | Respiratory epithelial cells | Medium | Protein (IHC) | HPA → |
Undetected expression · recommended negative controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| Adipose tissue | Adipocytes | Not detected | Protein (IHC) | HPA → |
| Cervix | Glandular cells | Not detected | Protein (IHC) | HPA → |
| Esophagus | Squamous epithelial cells | Not detected | Protein (IHC) | HPA → |
| Lymph node | Germinal center cells | Not detected | Protein (IHC) | HPA → |
| Oral mucosa | Squamous epithelial cells | Not detected | Protein (IHC) | HPA → |
Section 3
Advanced ARPC5 IHC Tips
Troubleshoot ARPC5 staining in paraffin sections by checking retrieval, compartment pattern and cell-level controls (UniProt O15511 localisation; HPA tissue IHC).
How should I adjust retrieval when ARPC5 staining is weak or diffuse?
Start with Tris-EDTA pH 9.0 heat-induced retrieval at 95–98 °C for 20 min (page retrieval setting). Run a no-primary control and a same-run positive section; the catalog image documents paraffin-embedded human colon, but its caption gives no retrieval conditions (M02096 tissue-IHC caption). If staining is weak, compare heat exposure around 20 min while holding antibody concentration and detection constant; citrate pH 6.0 is a fallback condition to test (standard IHC practice). Judge improvement by cellular detail and low background, using cytoplasmic staining as a reference pattern (HPA tissue IHC: general cytoplasmic expression).
Could fixation explain variable ARPC5 staining between paraffin blocks?
ARPC5-specific sensitivity to fixation is unknown from the supplied evidence; the selected paraffin-section caption does not state a fixative (M02096 tissue-IHC caption). Record fixation type and duration for each block, then compare sections processed with the same retrieval and detection conditions (standard IHC practice). Excessive or inconsistent fixation can change epitope accessibility in paraffin-section IHC, so test a small retrieval series before changing several variables together (standard IHC practice). Include a same-run reference section and no-primary control, and assess whether variation tracks processing batches rather than cellular compartments; cytoplasmic expression is the tissue-IHC reference pattern (HPA tissue IHC).
Which compartments should count as plausible ARPC5 staining in IHC?
Expect a predominantly cytoplasmic tissue-IHC pattern, while interpreting finer compartment detail cautiously (HPA tissue IHC: general cytoplasmic expression). ARPC5 is annotated in cytoplasm, cytoskeleton, cell projections and nucleus (UniProt O15511 localisation), and HPA subcellular imaging reports cytosol, vesicles, plasma membrane and cell junctions (HPA subcellular). On chromogenic sections, compare staining with cell boundaries and morphology at high magnification before calling a junctional or nuclear pattern (standard IHC interpretation). Score each compartment separately if it is reproducible across sections; isolated nuclear pigment or a uniform rim without cellular detail needs control-based review (standard IHC practice).
Can this stain distinguish ARPC5 isoforms or epitope accessibility?
Do not assign an isoform from a single chromogenic stain: the record lists isoforms 1 and 2, but supplies no antibody epitope or isoform-reactivity data (UniProt O15511 isoforms; supplied antibody evidence). ARPC5 has no annotated transmembrane segment or glycosylation sites, and its listed modified residue includes N-acetylserine at position 2 (UniProt O15511 topology, glycosylation and modified residues). Those annotations do not establish which epitope remains accessible after section processing (standard IHC interpretation). If isoform discrimination matters, obtain mapped epitope and cross-reactivity evidence, then validate a suitable reagent with orthogonal controls before assigning compartment-specific staining to either isoform (standard IHC practice).
How can I check ARPC5 localisation by IF alongside the IHC result?
Treat IF/ICC as a separate assay from this paraffin-section IHC workflow; HPA reports ARPC5 at cytosol, vesicles, plasma membrane and cell junctions in subcellular imaging (HPA subcellular). Multiplex ARPC5 with a marker identifying the expected cell type, and verify channel separation with single-stain controls (standard IF practice). Choose a fluorophore channel after checking tissue autofluorescence, favoring a cleaner, longer-wavelength channel when short-wavelength background is strong (standard IF practice). Because ARPC5 has no transmembrane segment and a cytosolic location is reported, use controlled permeabilisation to access intracellular epitopes; the antibody epitope itself is unspecified (UniProt O15511 topology; HPA subcellular; supplied antibody evidence).
What should I check when ARPC5 IHC shows widespread brown background?
First compare the stain with a no-primary control and inspect unstained or counterstained tissue for pigment and deposits (standard chromogenic IHC practice). Block endogenous peroxidase before peroxidase-based detection, optimise protein blocking and washes, and shorten chromogen development if the background rises across the section (standard chromogenic IHC practice). A diffuse cytoplasmic pattern can occur across several tissue types, so widespread signal alone does not establish nonspecificity (HPA tissue IHC: general cytoplasmic expression). Compare cellular detail, section edges and damaged areas with a same-run reference section; report uncertainty if background obscures cell boundaries or compartment assignment (standard IHC practice).
How should I score ARPC5 staining across cell populations? ⚠ ANSWER MARKED FOR VERIFICATION
Define the cell population and cytoplasmic compartment before scoring, since general cytoplasmic tissue expression is reported (HPA tissue IHC). For a chromogenic intensity readout, record the percentage of cells at intensity 0–3 and calculate an H-score from 0–300; alternatively report percentage positive cells using a prespecified threshold (standard IHC scoring practice). Normalise counts to the number of evaluable cells in the defined population, or report positive-cell density per mm² of evaluable tissue (standard IHC scoring practice). Keep exposure, counterstain, threshold and excluded damaged areas consistent across sections, and score distinct compartments separately when needed (standard IHC practice).
How can I separate true ARPC5 staining from artefact?
Give greatest weight to reproducible cellular staining in the expected population and a predominantly cytoplasmic pattern, which is the reported tissue-IHC profile (HPA tissue IHC). ARPC5 also has cytoskeletal, projection and nuclear annotations, but those alone do not validate a nuclear chromogenic signal (UniProt O15511 localisation; standard IHC interpretation). Review abrupt edge staining, necrotic regions and pigment that persists in a no-primary control; endogenous peroxidase can also create brown signal in peroxidase-based detection (standard chromogenic IHC practice). HPA labels its tissue antibody evidence Approved while warning of low staining–RNA consistency and possible recognition of more than one gene, so confirm consequential findings independently (HPA tissue IHC reliability description).
Boster reagents
Best ARPC5 / Actin-related protein 2/3 complex subunit 5 IHC Antibodies
ARPC5 images show IHC in paraffin-embedded human colon (M02096 image caption) and IF in human U-2 OS cells (M02096-1 image caption); both antibodies list human, mouse, and rat reactivity (catalog).
Real IHC data
Anti-p16 ARC ARPC5 Rabbit Monoclonal Antibody
Cat # M02096
Real IF data
Anti-ARPC5 Antibody (Center)
Cat # M02096-1
M02096 has a paraffin-embedded human colon IHC image and lists IHC, IF, and ICC applications (M02096 image caption; catalog). M02096-1 has a fixed, permeabilized human U-2 OS IF image and lists IF, but not IHC or ICC, as a tested application (M02096-1 image caption; catalog).
Which to pick: Choose monoclonal M02096 for tissue IHC: its image shows paraffin-embedded human colon, but the fixative is unreported (M02096 catalog; M02096 image caption). For cell IF, M02096-1 has a U-2 OS image; for ICC, M02096 lists that application (M02096-1 image caption; M02096 catalog). Both list human, mouse, and rat reactivity, while their pictured results are human samples (catalog; M02096 and M02096-1 image captions).
Each figure is that product's own IHC / IF validation image from its datasheet.
References
- UniProt Consortium. UniProt entry O15511 (ARPC5_HUMAN, Actin-related protein 2/3 complex subunit 5).
- Human Protein Atlas. ARPC5 tissue IHC expression (reliability: Approved).
- Human Protein Atlas. ARPC5 subcellular location (ICC-IF): Localized to the plasma membrane, cell junctions, vesicles and cytosol..
- Human Protein Atlas. ARPC5 antibody validation summary (2 antibodies).
- A comprehensively prognostic and immunological analysis of actin-related protein 2/3 complex subunit 5 in pan-cancer and identification in hepatocellular carcinoma. Frontiers in immunology 2022 — PMC9485570.
- Effects of hypothyroidism on expression of CRMP2B and ARPC5 during development of the rat frontal cortex. International journal of biological sciences 2013 — PMC3584917.
- ARPC5 acts as a potential prognostic biomarker that is associated with cell proliferation, migration and immune infiltrate in gliomas. BMC cancer 2023 — PMC10548738.
- Identification of Arp2/3 Complex Subunits as Prognostic Biomarkers for Hepatocellular Carcinoma. Frontiers in molecular biosciences 2021 — PMC8299467.
- PubMed PMID:9230079 — UniProt-cited evidence.
- PubMed PMID:9359840 — UniProt-cited evidence.
- PubMed PMID:16710414 — UniProt-cited evidence.
Immunohistochemistry protocol sheet for ARPC5 (Actin-related protein 2/3 complex subunit 5). Expected localisation Cytoplasmic staining observed (HPA tissue IHC); nuclear location annotated (UniProt). Antigen retrieval: Tris-EDTA pH 9.0 HIER, 95–98 °C, 20 min (rule: nuclear antigen). Validated IHC antibody: M02096 at 1:50 (the product's own IHC validation image is shown). Controls (HPA): positive Testis (Elongated or late spermatids); negative Adipose tissue. IHC workflow; specimen preparation unspecified. Protocol steps: 1. Sample: Paraffin-embedded human colon tissue; fixative not specified (datasheet M02096); 2. Fixation: Image fixative and duration unreported (datasheet M02096); 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-induced epitope retrieval in Tris-EDTA buffer, pH 9.0 , 20 min at 95–98 °C…; 6. Peroxidase block: 3% H2O2, 10 min , room temperature (standard); 7. Blocking: 10% normal serum of the secondary host, 30 min , room temperature (standard); 8. Primary antibody: Rabbit monoclonal (clone CCD-1) anti-ARPC5, 1:50 (datasheet M02096); 9. Primary incubation: Overnight at 4 °C (standard). Watch for: Fixation: Keep paraffin-section fixation consistent (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 M02096); Caveat: Antibody staining may reflect more than one gene (HPA tissue IHC); Isoform / epitope: 2 isoforms; epitope impact is unknown (UniProt); Regulation: Expression regulation unannotated (UniProt). Evidence: UniProt O15511, HPA tissue IHC · Approved, HPA ICC-IF · plasma membrane, cell junctions, vesicles and cytosol., Datasheet M02096, PMC9485570, PMC3584917. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/ARPC5/immunohistochemistry-ihc-guide.