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ARPC1B / Actin-related protein 2/3 complex subunit 1B · IHC design guide
Design Immunohistochemistry for ARPC1B
Plan ARPC1B IHC in paraffin sections using cytoplasmic staining in tonsil germinal center cells as a reference (HPA tissue IHC). This guide covers fixation consistency, the IHC-validated antibody at 2–5 μg/mL (datasheet A07192-2), and chromogenic detection.
Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
ARPC1B 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); nucleus annotated (UniProt) | |
| Staining pattern | Cytoplasmic; strongest in germinal center cells (HPA tissue IHC) | |
| Antigen retrieval | EDTA pH 8.0 HIER, heat-mediated (datasheet A07192-2) | |
| Positive control | Lymph node+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) | |
| Caveat | Staining and RNA expression have medium consistency (HPA tissue IHC) | |
| Regulation | No expression regulator annotated (UniProt) | |
| Isoform / epitope | No annotated isoforms; mature chain spans residues 2–372 (UniProt) |
Section 1
Recommended ARPC1B IHC & IF Protocols
The catalog antibody’s IHC-P protocol (datasheet A07192-2) is followed by published ARPC1B IHC protocols for glioblastoma (PMC10770598) and renal clear cell carcinoma (PMC10546172).
| Sample | Paraffin-embedded human tonsil tissue; fixative not specified (datasheet A07192-2) |
| Fixation | Image fixative and duration unreported (datasheet A07192-2); 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 A07192-2); 20 min, 95–100 °C (standard) |
| Peroxidase block | 3% H2O2, 10 min, room temperature (standard) |
| Blocking | 10% goat serum (datasheet A07192-2) |
| Primary antibody | Rabbit anti-ARPC1B, 2-5μg/ml (datasheet A07192-2) |
| Primary incubation | Overnight at 4 °C (datasheet A07192-2) |
| Detection | HRP-conjugated secondary, DAB chromogen (datasheet A07192-2) |
| Counterstain | Hematoxylin, blue, dehydrate and mount (standard) |
| Expected result | ARPC1B-positive staining in germinal center cells of lymph node (HPA tissue IHC: High). HPA tissue profile: Cytoplasmic expression in several tissues, most abundant in lymphoid tissues. No signal in the no-primary control. |
💡Decision noteStart with heat-mediated EDTA pH 8.0 retrieval (datasheet A07192-2); compare Tris/EDTA pH 9.0 for renal FFPE sections (PMC10546172).
Section 2
What Is the Expected ARPC1B Staining Pattern?
ARPC1B is a cytoplasmic, cytoskeletal protein that can also localize to the nucleus; it has no transmembrane segment (UniProt O15143). In paraffin-section IHC, expect mainly cytoplasmic staining, strongest in lymph-node and tonsil germinal-center cells (HPA: High). HPA describes cytoplasmic expression across several tissues, most abundant in lymphoid tissues; its tissue IHC reliability is Enhanced, with medium consistency between staining and RNA expression (HPA: tissue IHC).
| Strong cytoplasmic staining in germinal-center cells of lymph node or tonsil. | This matches the clearest tissue positive: both cell populations are rated High (HPA: tissue IHC). Compare intensity within the identified cells, since other cells in the same section need not show the same signal (HPA: cell-specific tissue IHC). |
| A predominantly membrane-bound or extracellular pattern, with little intracellular signal. | Treat this as a localization mismatch: ARPC1B is cytoplasmic and cytoskeletal, with a nuclear location also recorded, and has no transmembrane segment (UniProt O15143). Check morphology and detection background before interpreting it as specific staining (general IHC practice). |
| Strong signal confined to adipocytes or colon glandular cells. | Those cell populations are listed as Not detected (HPA: tissue IHC). Recheck cell identity, antibody specificity and endogenous chromogenic activity before assigning the signal to ARPC1B (general IHC practice); the HPA rating applies to the named cells, not every cell in those tissues. |
| Diffuse color across tissue and blank spaces, obscuring cell boundaries. | This does not resolve the expected cytoplasmic cell pattern (HPA: tissue IHC). Background can arise from detection chemistry, inadequate blocking or excessive antibody concentration (general IHC practice); compare a control processed without primary antibody. |
| No discernible signal in lymph-node or tonsil germinal-center cells. | This conflicts with the High staining recorded in those cells (HPA: tissue IHC), but one absent result does not establish absent protein. Check section integrity, retrieval and detection with suitable controls, then reassess the antibody conditions (general IHC practice). |
💡Expected ARPC1B appearanceCall a result positive when germinal-center cells show clear, predominantly cytoplasmic staining of high relative intensity (HPA: lymph node and tonsil, High); isolated strong staining of cells rated Not detected, or color lacking cellular boundaries, warrants investigation rather than a positive ARPC1B call (HPA: tissue IHC; general IHC practice).
| Tissue and cell choice | Lymph-node and tonsil germinal-center cells provide High-staining reference populations; appendix lymphoid tissue and bone-marrow hematopoietic cells are Medium (HPA: tissue IHC). Score the named cell population, because a whole-tissue average can hide that distinction. |
| Compartment | Cytoplasmic staining is the tissue IHC pattern (HPA: tissue IHC). UniProt also records cytoskeleton and nucleus, so a nuclear component alone is not grounds to reject a result; assess whether the overall cell and tissue pattern agrees (UniProt O15143). |
| Topology and processing | No transmembrane segment, signal peptide, propeptide or glycosylation sites are annotated (UniProt O15143). These annotations support an intracellular interpretation; they do not identify this antibody’s epitope or establish an antigen-retrieval condition. |
| Strength of validation | The tissue IHC assessment is Enhanced but has medium staining-to-RNA consistency (HPA: tissue IHC). HPA004832 is also rated Enhanced for IHC (HPA: antibody record); use those ratings as supporting evidence, not proof that every observed positive cell is specific. |
| IF/ICC: should its pattern match this IHC call? | HPA reports supported vesicle and cytosol localization in ICC-IF, while HPA004832 has Uncertain ICC validation (HPA: subcellular; antibody record). Those images inform localization, but they do not validate an IF/ICC protocol or override the paraffin-section tissue pattern. |
| Situation | Likely cause | Next action |
|---|---|---|
| Germinal-center cells are blank. | The result conflicts with their High HPA staining; a failed IHC workflow is one possibility (HPA: tissue IHC; general IHC practice). | Confirm the cells and section quality, then check retrieval, primary-antibody incubation and detection against working controls (general IHC practice). Do not infer ARPC1B absence from this slide alone. |
| Most cells look uniformly dark. | Uniform color obscures HPA’s cell-specific intensity differences and can reflect excessive antibody or chromogen development (HPA: tissue IHC; general IHC practice). | Compare a control without primary antibody, shorten development or optimize antibody concentration, and rescore only where cell boundaries remain readable (general IHC practice). |
| Unexpected staining appears in HPA-negative cell populations. | Adipocytes and colon glandular cells are rated Not detected (HPA: tissue IHC); cross-reactivity or endogenous detection activity is possible (general IHC practice). | Verify cell identity and inspect a no-primary control. If using peroxidase detection, assess the endogenous-enzyme blocking step, then compare a known-positive section (general IHC practice). |
| Only a crisp surface rim is visible. | A membrane-restricted result conflicts with the intracellular localization and lack of a transmembrane segment (UniProt O15143). | Inspect focus and morphology, compare a no-primary control, and repeat with adjusted staining conditions if the rim persists (general IHC practice). Avoid scoring it as the expected pattern. |
| A faint result appears in appendix or bone marrow. | The named lymphoid or hematopoietic cells are rated Medium, below the High germinal-center reference populations (HPA: tissue IHC). | Identify and score those cells separately; compare lymph-node or tonsil germinal-center cells in a matched run before judging assay sensitivity (HPA: tissue IHC; general IHC practice). |
| Nuclear or punctate staining prompts disagreement with the IHC call. | UniProt records a nuclear location, and HPA ICC-IF reports vesicles and cytosol; HPA004832 ICC validation is Uncertain (UniProt O15143; HPA: subcellular and antibody record). | For paraffin IHC, anchor the call to the cytoplasmic tissue pattern and expected cell types (HPA: tissue IHC). Document any additional compartment separately instead of treating ICC-IF localization as IHC validation. |
Experiment evidence
IHC & IF images showing ARPC1B staining
A07192-2 · IHCIHC analysis of ARPC1B using anti-ARPC1B antibody (A07192-2). ARPC1B was detected in a paraffin-embedded section of human tonsil tissue. Hea
HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000130429-ARPC1B/tissue/cerebral+cortex#imid_1652536
HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000130429-ARPC1B/tissue/colon#imid_1652462
HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000130429-ARPC1B/tissue/liver#imid_1652463
HPA · KidneyHuman Protein Atlas IHC, Kidney. Source: https://www.proteinatlas.org/ENSG00000130429-ARPC1B/tissue/kidney#imid_1652456
HPA · ICC-IF A-431Human Protein Atlas immunofluorescence, A-431, antibody HPA004832; green = ARPC1B, blue = nucleus, red = microtubules.
HPA · ICC-IF A-431Human Protein Atlas immunofluorescence, A-431, antibody HPA004832; green = ARPC1B, blue = nucleus, red = microtubules.
← swipe / scroll for more images →
Real IHC / IF data behind the staining-pattern expectations in this section.
Sample controls for ARPC1B IHC & IF
🧪Run tonsil first and look for staining in germinal center cells (HPA: High in tonsil germinal center cells). Use adipose tissue as a negative comparator, scoring adipocytes specifically (HPA: Not detected in adipocytes); on the tonsil slide, cells without specific signal should show counterstain without DAB deposition, although HPA does not identify a negative tonsillar cell type.
Positive control tissue: Lymph node (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 ARPC1B in A-431, U-251MG, U2OS, NIH 3T3, with annotated localisation: Vesicles (supported), Cytosol (supported) (HPA subcellular).
Technical controls: Include a no-primary (secondary-only) control, a concentration-matched nonimmune rabbit IgG control, and an ARPC1B-knockout specimen as a biological negative (caption: rabbit primary antibody). Quench endogenous peroxidase and assess background in tonsil before interpreting HRP–DAB staining (caption: tonsil section with peroxidase detection and DAB).
⚠️Feasibility: No target-specific fixation window or fixation effect is reported, and the selected A07192-2 tissue-IHC caption does not state a fixative (caption: fixative not stated). The caption reports staining in a paraffin-embedded tonsil section after heat-mediated EDTA retrieval at pH 8.0; dependence on that retrieval condition is unreported (caption: tonsil IHC). Frozen-section or tissue IF ease cannot be determined from these data; ICC-IF images support vesicular and cytosolic localization in the listed cell lines, while endogenous peroxidase is a practical background concern for tonsil HRP–DAB IHC (HPA: subcellular ICC-IF; caption: tonsil HRP–DAB IHC).
HPA tissue IHC evidence for ARPC1B
Comprehensive Human Protein Atlas IHC scoring per tissue (reliability: Enhanced — Medium consistency between antibody staining and RNA expression data.). 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 |
|---|---|---|---|---|
| Lymph node | Germinal center cells | High | Protein (IHC) | HPA → |
| Tonsil | Germinal center cells | High | Protein (IHC) | HPA → |
| Appendix | Lymphoid tissue | Medium | Protein (IHC) | HPA → |
| Bone marrow | Hematopoietic cells | Medium | Protein (IHC) | HPA → |
| Skin | Fibroblasts | Medium | 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 → |
| Bronchus | Respiratory epithelial cells | Not detected | Protein (IHC) | HPA → |
| Caudate | Glial cells | Not detected | Protein (IHC) | HPA → |
| Cerebellum | Cells in granular layer | Not detected | Protein (IHC) | HPA → |
Section 3
Advanced ARPC1B IHC Tips
Troubleshoot ARPC1B staining in paraffin sections by checking retrieval, cell type and compartment against the available tissue evidence.
How should I retrieve ARPC1B when staining is weak in paraffin sections?
Start with heat-mediated retrieval in EDTA at pH 8.0 (datasheet A07192-2). The selected tonsil image used that retrieval before incubation with 2 μg/ml primary antibody overnight at 4°C, so it provides a practical comparison condition (caption A07192-2). If staining is weak, check heating uniformity, cooling and section adhesion before changing retrieval conditions (standard IHC practice). Run a tonsil control alongside the study sections because germinal center cells show high staining (HPA tissue IHC). Compare changes at matched antibody concentration and DAB development time, and watch for increased background (standard IHC practice).
Could fixation explain weak ARPC1B staining in my paraffin sections?
Target-specific sensitivity to fixation is unknown: the selected paraffin-section caption does not state a fixative (caption A07192-2). Record the fixative, fixation duration and processing history for each block before comparing staining across specimens (standard IHC practice). If archived sections stain unevenly, compare blocks with similar handling and include a concurrently stained positive control (standard IHC practice). The tonsil example establishes retrieval in EDTA at pH 8.0 and primary incubation overnight at 4°C, but supplies no fixation test (caption A07192-2). Treat any improvement after changing fixation conditions as an experimental finding requiring matched controls, rather than an established ARPC1B effect (standard IHC practice).
Which ARPC1B staining compartments should I expect in chromogenic IHC?
Expect predominantly cytoplasmic staining across tissues, with stronger signal in lymphoid tissue (HPA tissue IHC). ARPC1B is associated with the cytoplasmic cytoskeleton and is also annotated in the nucleus, while cell imaging supports cytosol and vesicles (UniProt O15143; HPA subcellular). Score nuclear and cytoplasmic staining separately rather than treating every brown compartment as equivalent (standard IHC practice). High staining in tonsil and lymph-node germinal center cells provides a useful reference pattern (HPA tissue IHC). If an apparently strong signal is confined to tissue edges or extracellular deposits, reassess section quality and background before assigning localisation (standard IHC practice).
How can I investigate unexpected ARPC1B staining when the antibody epitope is unspecified?
The supplied record lists 0 annotated isoforms, 0 glycosylation sites and 0 modified residues; it does not identify this antibody’s epitope (UniProt O15143; supplied product evidence). ARPC1B has no transmembrane segment, so an exclusively membrane-rim pattern needs further scrutiny (UniProt O15143 topology). Review the antibody’s stated immunogen or epitope documentation before drawing conclusions about epitope exposure (standard IHC practice). Compare the same staining run across a high-expression germinal center control and a tissue or cell population reported as undetected (HPA tissue IHC). If available, use an independently validated antibody or a specific loss-of-target control to test whether the unexpected pattern tracks ARPC1B (standard IHC practice).
How should I investigate ARPC1B localisation with IF after observing IHC staining?
Use IF as a separate localisation check, while retaining the chromogenic tonsil section as the IHC reference (caption A07192-2). Multiplex ARPC1B with a validated germinal center cell marker when examining tonsil, since those cells show high tissue staining (HPA tissue IHC; standard IF practice). Choose fluorophores after inspecting unstained tissue for autofluorescence, and use separate channels and single-label controls to assess bleed-through (standard IF practice). Permeabilise cells to reach cytosolic and potential nuclear epitopes; ARPC1B has no transmembrane segment, and its precise antibody epitope is unreported (UniProt O15143; supplied product evidence; standard IF practice). Compare cytosolic or vesicular signal with the supported cell-imaging locations before interpreting additional compartments (HPA subcellular).
What should I check when ARPC1B DAB staining appears widespread or hazy?
Compare background with a no-primary control and inspect whether deposits follow tissue edges, folds or damaged areas (standard IHC practice). The selected tonsil image used 10% goat serum blocking and a peroxidase-linked secondary, followed by DAB detection (caption A07192-2). Include an appropriate endogenous peroxidase block and assess it with controls before attributing diffuse brown signal to ARPC1B (standard IHC practice). Reduce excess primary antibody or DAB development only while keeping a known positive section in the run (standard IHC practice). Cytoplasmic staining is reported across several tissues, but its abundance differs by cell population, so uniform haze warrants investigation (HPA tissue IHC).
How should I quantify ARPC1B IHC across sections with different cell mixtures? ⚠ ANSWER MARKED FOR VERIFICATION
Define the cell population and compartment before scoring, then apply the same threshold and DAB development conditions across slides (standard IHC practice). Report the percentage of positive cells and an intensity-based H-score, or positive-cell density per mm² for spatial comparisons (standard IHC practice). Normalise to the number or area of evaluable cells of the same type, rather than total section area when cellular composition varies (standard IHC practice). Keep germinal center cells distinct from surrounding populations because they show high ARPC1B staining in tonsil and lymph node (HPA tissue IHC). Record excluded folds, necrosis and edge regions, and use matched control sections to monitor staining-run variation (standard IHC practice).
How can I distinguish genuine ARPC1B positivity from staining artefacts?
Prioritise cell identity and compartment: strong germinal center staining in tonsil or lymph node fits the tissue profile, and cytoplasmic staining is commonly reported (HPA tissue IHC). Cytosol and vesicles are supported imaging locations, while nuclear localisation is annotated separately, so record unexpected patterns rather than merging them into one score (HPA subcellular; UniProt O15143). Check an undetected reference population, such as adipocytes, within the limits of the reported tissue pattern (HPA tissue IHC). Reject edge-only staining, deposits over necrosis and signal retained in a no-primary control as evidence of specific positivity (standard IHC practice). Investigate endogenous peroxidase activity before accepting diffuse DAB signal, especially when cell boundaries are unclear (standard IHC practice).
Boster reagents
Best ARPC1B / Actin-related protein 2/3 complex subunit 1B IHC Antibodies
The IHC-validated ARPC1B antibody has a paraffin-section image from human tonsil (catalog image caption); the catalog also lists human, mouse and rat reactivity (catalog: A07192-2).
Real IHC data
Anti-ARP2/3 subunit 1B/ARPC1B Antibody ®
Cat # A07192-2
A07192-2 is the only SKU shown and has an IHC image from a paraffin-embedded human tonsil section (catalog image caption). Its listed applications include IHC, and its listed reactivity is human, mouse and rat (catalog: A07192-2).
Which to pick: Choose A07192-2 for paraffin-section IHC: its own image shows human tonsil with EDTA pH 8.0 retrieval and 2 μg/ml primary antibody (catalog image caption). Its reactivity list includes human, mouse and rat, but the supplied IHC image documents human tissue only; it is rabbit-hosted, with clonality and fixative unreported (catalog: A07192-2; catalog image caption). No IF/ICC-validated SKU or IF image is supplied, so there is no supported IF/ICC pick (catalog: A07192-2).
Each figure is that product's own IHC / IF validation image from its datasheet.
References
- UniProt Consortium. UniProt entry O15143 (ARC1B_HUMAN, Actin-related protein 2/3 complex subunit 1B).
- Human Protein Atlas. ARPC1B tissue IHC expression (reliability: Enhanced).
- Human Protein Atlas. ARPC1B subcellular location (ICC-IF): Localized to vesicles and cytosol..
- Human Protein Atlas. ARPC1B antibody validation summary (1 antibodies).
- High expression of ARPC1B correlates with immune infiltration and poor outcomes in glioblastoma. Biochemistry and biophysics reports 2024 — PMC10770598.
- Dual role of ARPC1B in regulating the network between tumor-associated macrophages and tumor cells in glioblastoma. Oncoimmunology 2022 — PMC8803105.
- ARPC1B is a novel prognostic biomarker for kidney renal clear cell carcinoma and correlates with immune infiltration. Frontiers in molecular biosciences 2023 — PMC10546172.
- Pan-cancer identified ARPC1B as a promising target for tumor immunotherapy and prognostic biomarker, particularly in READ. Heliyon 2024 — PMC11059418.
- PubMed PMID:9230079 — UniProt-cited evidence.
- PubMed PMID:12853948 — UniProt-cited evidence.
- PubMed PMID:15489334 — UniProt-cited evidence.
Immunohistochemistry protocol sheet for ARPC1B (Actin-related protein 2/3 complex subunit 1B). Expected localisation Cytoplasmic staining observed (HPA tissue IHC); nucleus annotated (UniProt). Antigen retrieval: EDTA pH 8.0 HIER, heat-mediated (datasheet A07192-2). Validated IHC antibody: A07192-2 at 2-5μg/ml (the product's own IHC validation image is shown). Controls (HPA): positive Lymph node (Germinal center cells); negative Adipose tissue. IHC-P workflow on paraffin sections; fixative unspecified. Protocol steps: 1. Sample: Paraffin-embedded human tonsil tissue; fixative not specified (datasheet A07192-2); 2. Fixation: Image fixative and duration unreported (datasheet A07192-2); 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 A07192-2); 20 min, 95–100 °C (standard); 6. Peroxidase block: 3% H2O2, 10 min , room temperature (standard); 7. Blocking: 10% goat serum (datasheet A07192-2); 8. Primary antibody: Rabbit anti-ARPC1B, 2-5μg/ml (datasheet A07192-2); 9. Primary incubation: Overnight at 4 °C (datasheet A07192-2). Watch for: Fixation: Keep fixation consistent across sections. (standard IHC practice; not target-specific); Caveat: Staining and RNA expression have medium consistency (HPA tissue IHC); Isoform / epitope: No annotated isoforms; mature chain spans residues 2–372 (UniProt); Regulation: No expression regulator annotated (UniProt). Evidence: UniProt O15143, HPA tissue IHC · Enhanced, HPA ICC-IF · vesicles and cytosol., Datasheet A07192-2, PMC10770598, PMC10546172. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/ARPC1B/immunohistochemistry-ihc-guide.