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SMARCA2 / SWI/SNF-related matrix-associated actin-dependent regulator of chromatin subfamily A member 2 · IHC design guide
Design Immunohistochemistry for SMARCA2
Plan paraffin SMARCA2 IHC around nuclear staining, using breast glandular cells as a high-staining reference (HPA tissue IHC). Start the IHC-validated antibody at 2–5 μg/ml (datasheet A01888-3) and score staining by cell type (standard IHC practice).
Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
SMARCA2 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 | Nuclear staining in tissue (HPA tissue IHC) | |
| Staining pattern | Widespread nuclear staining across cell types (HPA tissue IHC) | |
| Antigen retrieval | EDTA pH 8.0 HIER, heat-mediated (datasheet A01888-3) | |
| Positive control | Bone marrow+4 more · see all | |
| Negative control | Soft tissue |
Important caveats
Reasons your staining may differ from the expected pattern.
| Fixation | Keep fixation consistent across sections. (standard IHC practice; not target-specific) | |
| Caveat | High hematopoietic-cell signal may confound bulk scoring (HPA tissue IHC) | |
| Regulation | Broad tissue expression (HPA tissue IHC) | |
| Isoform / epitope | Two isoforms, Long and Short; epitope coverage unknown (UniProt) |
Section 1
Recommended SMARCA2 IHC & IF Protocols
The catalog antibody uses EDTA pH 8.0 retrieval (datasheet: A01888-3). The published IHC protocols below cover lung, ovarian, and pancreatic tumors (PMC12439815; PMC4697871; PMC11585460).
| Sample | Paraffin-embedded human appendix adenocarcinoma tissue; fixative not specified (datasheet A01888-3) |
| Fixation | Image fixative and duration unreported (datasheet A01888-3); 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 A01888-3); 20 min, 95–100 °C (standard) |
| Peroxidase block | 3% H2O2, 10 min, room temperature (standard) |
| Blocking | 10% goat serum (datasheet A01888-3) |
| Primary antibody | Rabbit anti-SMARCA2, 2-5 μg/ml (datasheet A01888-3) |
| Primary incubation | Overnight at 4 °C (datasheet A01888-3) |
| Detection | HRP-conjugated secondary, DAB chromogen (datasheet A01888-3) |
| Counterstain | Hematoxylin, blue, dehydrate and mount (standard) |
| Expected result | SMARCA2-positive staining in hematopoietic cells of bone marrow (HPA tissue IHC: High). HPA tissue profile: Ubiquitous nuclear expression. No signal in the no-primary control. |
💡Decision noteStart with heat-mediated EDTA pH 8.0 retrieval (datasheet: A01888-3); an ovarian tumor study used this buffer with 30 min steaming (PMC4697871: Immunohistochemistry).
Section 2
What Is the Expected SMARCA2 Staining Pattern?
SMARCA2 is a nuclear chromatin-remodeling protein with no transmembrane segment (UniProt P51531). In paraffin-section IHC, expect staining chiefly in nuclei across many cell types: HPA describes ubiquitous nuclear expression, with high staining in several specified populations (HPA tissue IHC). The tissue profile has Enhanced reliability, reflecting consistency between antibody staining and RNA expression (HPA tissue IHC).
| Distinct nuclear chromogen in breast glandular cells, bone-marrow hematopoietic cells, or caudate neurons. | This matches the expected compartment (UniProt P51531; HPA tissue IHC). HPA reports High staining in each named population; assess nuclei within the relevant cell population rather than assigning one intensity to every cell in the section (HPA tissue IHC). |
| Predominantly cytoplasmic or membranous chromogen, with little nuclear staining. | Treat this as a questionable IHC result: the expected tissue pattern is nuclear (HPA tissue IHC; UniProt P51531). Review morphology, background, and detection controls before calling it SMARCA2. HPA reports uncertain additional plasma-membrane and vesicle locations in ICC-IF; those observations do not establish a dominant membranous IHC pattern (HPA subcellular). |
| Strong apparent staining in soft-tissue peripheral nerve while nearby expected nuclear staining is absent. | HPA lists peripheral nerve in soft tissue as Not detected (HPA tissue IHC). Investigate cross-reactivity or endogenous detection activity (general IHC practice). Because HPA describes broadly distributed nuclear expression, judge the identified cell population and staining compartment rather than treating an entire tissue as negative (HPA tissue IHC). |
| Haze or chromogen deposited broadly across nuclei, cytoplasm, and extracellular areas. | Diffuse staining that ignores cell boundaries and the expected nuclear compartment cannot by itself establish SMARCA2 positivity (HPA tissue IHC; general IHC practice). Assess a detection-only control and local tissue background; blocking, washes, and chromogen development are general workflow variables (general IHC practice). |
| No nuclear signal in a section containing identifiable breast glandular cells or bone-marrow hematopoietic cells. | Those populations are reported as High by HPA, so a blank result warrants a technical check (HPA tissue IHC). Confirm that the expected cells are present, then check the IHC-validated antibody and detection run (general IHC practice). A blank slide alone does not establish biological absence. |
💡Expected SMARCA2 appearanceCall a convincing IHC positive when identifiable cells show predominantly nuclear chromogen, potentially High in HPA-listed positive populations; widespread cytoplasmic or membranous staining without a nuclear pattern is suspect (HPA tissue IHC; UniProt P51531).
| Tissue and cell population | HPA reports High staining in breast glandular cells, cerebral-cortex glia, and several other listed populations; adrenal glandular cells are Low, and soft-tissue peripheral nerve is Not detected (HPA tissue IHC). Use the specific cell population when interpreting intensity. |
| Antibody evidence | HPA lists IHC as Supported for HPA029981 and Enhanced for CAB037276 (HPA antibodies). These are antibody-specific validation labels; confirm which antibody was used before applying either label to a slide. The tissue-profile reliability is separately Enhanced (HPA tissue IHC). |
| Isoforms and epitope | UniProt lists Long and Short isoforms (UniProt P51531). The supplied records do not locate the catalog antibody's epitope or establish whether it recognizes both isoforms. Interpret a weak result against antibody documentation and a known-positive tissue, without assigning an isoform-specific staining pattern. |
| IF/ICC: where should signal appear? | HPA supports the nucleoplasm as the main location in ICC-IF images (HPA subcellular). Vesicle and plasma-membrane locations are marked uncertain, so do not use them as the primary positive criterion (HPA subcellular). IF/ICC interpretation belongs with its own guide page. |
| Situation | Likely cause | Next action |
|---|---|---|
| Expected positive population has no nuclear chromogen. | The cells may be absent from the section, or the staining or detection run may have failed (general IHC practice). | Identify the cells on the counterstain; compare a tissue with HPA-reported High staining, then review antibody dilution, retrieval, and detection against the validated IHC workflow (HPA tissue IHC; general IHC practice). |
| Signal is mainly cytoplasmic or at cell borders. | This conflicts with the expected nuclear IHC distribution (UniProt P51531; HPA tissue IHC). | Check nuclear morphology and a detection-only control; repeat with the IHC-validated antibody if the compartment remains inconsistent (general IHC practice). Do not promote uncertain ICC-IF membrane localization to an IHC acceptance rule (HPA subcellular). |
| Soft-tissue peripheral nerve appears strongly positive. | The identified population is reported Not detected by HPA; cross-reactivity or endogenous detection activity is possible (HPA tissue IHC; general IHC practice). | Verify the cell identity and nuclear location, inspect a detection-only control, and compare the result with an HPA-listed positive population (HPA tissue IHC; general IHC practice). |
| Chromogen covers much of the section as diffuse background. | Non-specific binding, incomplete washes, or excessive chromogen development can obscure compartments (general IHC practice). | Inspect controls and review blocking, wash steps, detection reagent, and development time under the established IHC workflow (general IHC practice). Score only interpretable nuclei. |
| Staining is weak in adrenal glandular cells. | HPA reports this population as Low (HPA tissue IHC). | Check a separate HPA-listed High population before changing assay conditions; compare nuclear staining within the same identified cell type and avoid treating low intensity alone as run failure (HPA tissue IHC; general IHC practice). |
| Results differ between antibodies or staining runs. | HPA reports different IHC validation levels for HPA029981 and CAB037276; the supplied records do not establish a shared epitope or identical isoform recognition (HPA antibodies; UniProt P51531). | Record the antibody identity, apply its validated IHC workflow consistently, and compare nuclear staining in the same HPA-listed positive cell population (HPA antibodies; HPA tissue IHC; general IHC practice). |
Experiment evidence
IHC & IF images showing SMARCA2 staining
A01888-3 · IHCIHC analysis of SMARCA2/BRM using anti-SMARCA2/BRM antibody (A01888-3). SMARCA2/BRM was detected in a paraffin-embedded section of human app
A01888-3 · IHCIHC analysis of SMARCA2/BRM using anti-SMARCA2/BRM antibody (A01888-3). SMARCA2/BRM was detected in a paraffin-embedded section of human bre
A01888-3 · IFIF analysis of SMARCA2/BRM and Tubulin beta using anti-SMARCA2/BRM antibody (A01888-3) and anti-Tubulin beta antibody (M05613-4). SMARCA2/BR
M01888 · IFIF analysis of SMARCA2/BRM using anti-SMARCA2/BRM antibody (M01888). SMARCA2/BRM was detected in immunocytochemical section of A549 cells. E
M01888-1 · IFIF analysis of SMARCA2/BRM using anti-SMARCA2/BRM antibody (M01888-1). SMARCA2/BRM was detected in immunocytochemical section of A549 cells.
HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000080503-SMARCA2/tissue/cerebral+cortex#imid_8333280
HPA · Parathyroid glandHuman Protein Atlas IHC, Parathyroid gland. Source: https://www.proteinatlas.org/ENSG00000080503-SMARCA2/tissue/parathyroid+gland#imid_8362351
HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000080503-SMARCA2/tissue/colon#imid_8362394
HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000080503-SMARCA2/tissue/liver#imid_8362387
HPA · ICC-IF A-431Human Protein Atlas immunofluorescence, A-431, antibody HPA029981; green = SMARCA2, blue = nucleus, red = microtubules.
HPA · ICC-IF A-431Human Protein Atlas immunofluorescence, A-431, antibody HPA029981; green = SMARCA2, blue = nucleus, red = microtubules.
← swipe / scroll for more images →
Real IHC / IF data behind the staining-pattern expectations in this section.
Sample controls for SMARCA2 IHC & IF
🧪Run breast first and expect nuclear staining in glandular cells (HPA: High in breast glandular cells; UniProt P51531: nucleus). Use soft tissue peripheral nerve as the negative tissue (HPA: Not detected in peripheral nerve); on the breast slide, cells without nuclear signal provide a local background reference, but no specific breast cell type is established as SMARCA2-negative by the supplied rows.
Positive control tissue: Bone marrow (Hematopoietic cells, HPA High)
Negative control tissue: Soft tissue (HPA Not detected)
ICC-IF cell lines (HPA subcellular resource): HPA ICC-IF images show SMARCA2 in A-431, U-251MG, U2OS, KOLF2.1J, with annotated localisation: Nucleoplasm (supported) (HPA subcellular).
Technical controls: Include a no-primary, secondary-only control and a concentration-matched rabbit IgG isotype control for the rabbit catalog antibody; a SMARCA2 knockout specimen, if available, provides a biological negative (A01888-3 caption: rabbit primary and anti-rabbit secondary; standard IHC practice). Quench endogenous peroxidase and check for nonspecific DAB deposition in the breast section (A01888-3 caption: HRP/DAB detection; standard IHC practice).
⚠️Feasibility: No target-specific fixation window or fixation effect is reported, and the selected A01888-3 paraffin-section caption does not state a fixative (A01888-3 caption: fixative unreported). The demonstrated IHC procedure used heat retrieval in EDTA at pH 8.0; whether retrieval is required, or frozen sections are easier, is unreported (A01888-3 caption). IF/ICC shows supported nucleoplasmic localization, but the supplied evidence does not establish that IF is easier; in breast, score glandular nuclei and exclude luminal DAB deposits from nuclear scoring (HPA: nucleoplasm supported; HPA: High in breast glandular cells; standard IHC practice).
HPA tissue IHC evidence for SMARCA2
Comprehensive Human Protein Atlas IHC scoring per tissue (reliability: Enhanced — High 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 |
|---|---|---|---|---|
| Bone marrow | Hematopoietic cells | High | Protein (IHC) | HPA → |
| Breast | Glandular cells | High | Protein (IHC) | HPA → |
| Caudate | Neuronal cells | High | Protein (IHC) | HPA → |
| Cerebellum | Cells in granular layer | High | Protein (IHC) | HPA → |
| Cerebral cortex | Glial cells | High | Protein (IHC) | HPA → |
Undetected expression · recommended negative controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| Soft tissue | Peripheral nerve | Not detected | Protein (IHC) | HPA → |
Section 3
Advanced SMARCA2 IHC Tips
Troubleshoot chromogenic SMARCA2 staining by checking nuclear localisation, retrieval and controls before comparing signal across tissue regions.
Which retrieval condition should I try first for weak SMARCA2 staining?
Use heat-mediated retrieval in EDTA at pH 8.0 for paraffin sections (datasheet A01888-3). The selected tissue image used this condition before overnight incubation with 2 μg/ml antibody at 4°C, so keep those steps consistent while investigating weak staining (datasheet A01888-3). Check that sections remain covered by retrieval buffer during heating and cool consistently before blocking, since uneven retrieval can produce patchy nuclear signal (standard IHC practice; UniProt P51531 localisation). If staining remains weak, compare a small set of retrieval times or a different buffer on matched sections, recording nuclear signal and tissue damage separately (standard IHC practice).
Could fixation explain inconsistent SMARCA2 nuclear staining?
The selected paraffin-section caption does not report a fixative, so SMARCA2-specific fixation sensitivity is unknown for this antibody (datasheet A01888-3). Record the fixative, fixation duration, specimen thickness and processing history for each section before comparing staining intensity (standard IHC practice). Compare matched sections processed together, using the same EDTA pH 8.0 retrieval and 2 μg/ml primary concentration to reduce procedural differences (datasheet A01888-3; standard IHC practice). If nuclear staining varies across a specimen, inspect morphology and internal nuclei before changing antibody concentration; poor preservation or processing can complicate interpretation (standard IHC practice; UniProt P51531 localisation).
How should I assess cytoplasmic or membranous SMARCA2 staining?
Score convincing SMARCA2 staining in nuclei, because UniProt places the protein in the nucleus and HPA reports ubiquitous nuclear expression (UniProt P51531 localisation; HPA tissue IHC). HPA further supports nucleoplasmic localisation in cell images, while its vesicle and plasma-membrane assignments are uncertain (HPA subcellular). Compare suspected extranuclear signal with an adjacent section lacking primary antibody and with well-preserved nuclei in the same specimen (standard IHC practice). If staining is mainly cytoplasmic or membranous, review retrieval, antibody concentration and DAB development before calling it SMARCA2-positive; the protein has no transmembrane segment (UniProt P51531 topology; standard IHC practice).
Can this antibody distinguish the long and short SMARCA2 isoforms?
Do not assign an isoform from chromogenic staining alone: the record lists Long and Short isoforms, but the selected caption does not identify the antibody epitope (UniProt P51531 isoforms; datasheet A01888-3). Ask whether epitope mapping shows a sequence shared by both isoforms before treating nuclear staining as a combined SMARCA2 readout (standard antibody-validation practice; UniProt P51531 isoforms). SMARCA2 also has annotated modified residues and several domains, including a bromodomain, so sequence-level epitope information matters when comparing reagents (UniProt P51531 modifications and domains). Validate any proposed isoform-specific interpretation with an independent assay that resolves the isoforms (standard antibody-validation practice).
How should I investigate a SMARCA2 signal in a multiplex IF experiment?
Treat IF as a separate application from this antibody’s paraffin-section chromogenic evidence, which used EDTA pH 8.0 retrieval and DAB detection (datasheet A01888-3). For a multiplex check, pair nuclear SMARCA2 with a validated marker for the expected cell population, such as glandular cells in breast tissue, and assess both channels in the same cells (HPA tissue IHC; standard IF practice). Choose a fluorophore channel with low measured tissue autofluorescence and include single-channel controls to check bleed-through (standard IF practice). SMARCA2 has no transmembrane segment; access to its nuclear epitope requires appropriate permeabilisation in intact cells, with conditions checked against nuclear morphology (UniProt P51531 topology and localisation; standard IF practice).
What should I check when DAB appears throughout the section?
First distinguish diffuse DAB deposit from discrete nuclear staining, the expected SMARCA2 pattern (UniProt P51531 localisation; standard IHC practice). The selected image used 10% goat-serum blocking, 2 μg/ml primary antibody overnight at 4°C, and a peroxidase-linked secondary for 30 minutes at 37°C (datasheet A01888-3). Check a section without primary antibody, peroxidase blocking, washing and DAB development time when background obscures nuclei (standard chromogenic IHC practice). Compare any adjustment on adjacent sections and retain internal nuclear staining as a reference, since broadly expressed nuclear SMARCA2 can otherwise be mistaken for nonspecific signal (HPA tissue IHC; standard IHC practice).
How should I quantify heterogeneous SMARCA2 IHC staining? ⚠ ANSWER MARKED FOR VERIFICATION
Define the cell population and regions to score before examining treatment groups, then count nuclear rather than cytoplasmic or DAB haze signal (UniProt P51531 localisation; standard IHC quantification practice). Report the percentage of positive nuclei and, when intensity is reproducible, a nuclear H-score across prespecified intensity categories (standard IHC quantification practice). Normalise each result to the number of evaluable nuclei in the relevant cell population; for spatial counts, report positive nuclei per mm² of evaluable tissue (standard IHC quantification practice). Keep retrieval, staining batch, exposure to DAB and scoring thresholds consistent, and document excluded damaged or necrotic regions (standard IHC quantification practice).
How can I distinguish true SMARCA2 positivity from staining artefact?
A credible positive result is nuclear staining in preserved cells, consistent with SMARCA2’s nuclear localisation and HPA’s broadly nuclear tissue pattern (UniProt P51531 localisation; HPA tissue IHC). Check whether the stained cells match the annotated population: HPA reports high signal in breast glandular cells but no detected signal in peripheral nerve within soft tissue (HPA tissue IHC). Treat staining confined to section edges, necrotic areas or the wrong compartment as suspect, and compare adjacent viable regions (standard IHC interpretation practice; UniProt P51531 localisation). A no-primary control helps identify endogenous peroxidase or detection-system signal; interpret loss of nuclear staining alongside tissue preservation and internal controls (standard chromogenic IHC practice).
Boster reagents
Best SMARCA2 / SWI/SNF-related matrix-associated actin-dependent regulator of chromatin subfamily A member 2 IHC Antibodies
A01888-3 has human paraffin-section IHC images (catalog: IHC captions); all three have human A549-cell IF images (catalog: IF captions); A01888-3 also lists mouse and rat reactivity (catalog: reactivity).
Real IHC data
Anti-SMARCA2/BRM Antibody ®
Cat # A01888-3
Real IF data
Anti-SMARCA2/BRM Antibody ® (monoclonal, 3G3)
Cat # M01888
Real IF data
Anti-SMARCA2/BRM Antibody ® (monoclonal, 4E8)
Cat # M01888-1
A01888-3 shows IHC in human appendix adenocarcinoma, breast cancer, colon adenocarcinoma and diffuse large B-cell lymphoma paraffin sections, plus IF in A549 cells (catalog: A01888-3 image captions). M01888 (clone 3G3) and M01888-1 (clone 4E8) each show IF in A549 cells and list IF/ICC, but do not list IHC (catalog: applications and IF captions).
Which to pick: Choose A01888-3 for human paraffin-section chromogenic IHC: its own caption reports EDTA pH 8.0 heat retrieval, 2 μg/ml primary antibody and HRP/DAB detection; the fixative is unreported (catalog: A01888-3 IHC caption). For human IF/ICC, choose M01888 or M01888-1, which are mouse monoclonals with their own A549-cell IF images (catalog: hosts, clones, applications and IF captions). For mouse or rat samples, A01888-3 is the only listed reactive option; its documented IHC images are from human paraffin sections (catalog: reactivity and A01888-3 IHC captions).
Each figure is that product's own IHC / IF validation image from its datasheet.
References
- UniProt Consortium. UniProt entry P51531 (SMCA2_HUMAN, SWI/SNF-related matrix-associated actin-dependent regulator of chromatin subfamily A member 2).
- Human Protein Atlas. SMARCA2 tissue IHC expression (reliability: Enhanced).
- Human Protein Atlas. SMARCA2 subcellular location (ICC-IF): Mainly localized to the nucleoplasm. In addition localized to the plasma membrane and vesicles..
- Human Protein Atlas. SMARCA2 antibody validation summary (2 antibodies).
- SMARCA2 Deficiency While Preserving SMARCA4 in Lung Adenocarcinoma Combined with Abnormal β-Catenin Expression. Cancer management and research 2025 — PMC12439815.
- Concomitant loss of SMARCA2 and SMARCA4 expression in small cell carcinoma of the ovary, hypercalcemic type. Modern pathology : an official journal of the United States and Canadian Academy of Pathology, Inc 2016 — PMC4697871.
- SWI/SNF Complex-Deficient Undifferentiated Carcinoma of the Pancreas: Clinicopathologic and Genomic Analysis. Modern pathology : an official journal of the United States and Canadian Academy of Pathology, Inc 2024 — PMC11585460.
- Re-assigning the histologic identities of COV434 and TOV-112D ovarian cancer cell lines. Gynecologic oncology 2021 — PMC10039450.
- PubMed PMID:8223438 — UniProt-cited evidence.
- PubMed PMID:8208605 — UniProt-cited evidence.
- PubMed PMID:15164053 — UniProt-cited evidence.
Immunohistochemistry protocol sheet for SMARCA2 (SWI/SNF-related matrix-associated actin-dependent regulator of chromatin subfamily A member 2). Expected localisation Nuclear staining in tissue (HPA tissue IHC). Antigen retrieval: EDTA pH 8.0 HIER, heat-mediated (datasheet A01888-3). Validated IHC antibody: A01888-3 at 2-5 μg/ml (the product's own IHC validation image is shown). Controls (HPA): positive Bone marrow (Hematopoietic cells); negative Soft tissue. IHC-P workflow on paraffin sections; fixative unspecified. Protocol steps: 1. Sample: Paraffin-embedded human appendix adenocarcinoma tissue; fixative not specified (datasheet A01888-3); 2. Fixation: Image fixative and duration unreported (datasheet A01888-3); 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 A01888-3); 20 min, 95–100 °C (standard); 6. Peroxidase block: 3% H2O2, 10 min , room temperature (standard); 7. Blocking: 10% goat serum (datasheet A01888-3); 8. Primary antibody: Rabbit anti-SMARCA2, 2-5 μg/ml (datasheet A01888-3); 9. Primary incubation: Overnight at 4 °C (datasheet A01888-3). Watch for: Fixation: Keep fixation consistent across sections. (standard IHC practice; not target-specific); Caveat: High hematopoietic-cell signal may confound bulk scoring (HPA tissue IHC); Isoform / epitope: Two isoforms, Long and Short; epitope coverage unknown (UniProt); Regulation: Broad tissue expression (HPA tissue IHC). Evidence: UniProt P51531, HPA tissue IHC · Enhanced, HPA ICC-IF · Mainly localized to the nucleoplasm. In addition localized to the…, Datasheet A01888-3, PMC12439815, PMC4697871. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/SMARCA2/immunohistochemistry-ihc-guide.