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BCAS1 / Breast carcinoma-amplified sequence 1 · IHC design guide
Design Immunohistochemistry for BCAS1
Plan chromogenic BCAS1 IHC-P with the human-reactive catalog antibody, starting at 5 μg/mL (datasheet: IHC-P). Assess cytoplasmic staining in prostate glandular cells (HPA tissue IHC) alongside a negative control (standard IHC practice).
Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
BCAS1 Immunohistochemistry Experimental Design Guide
Expected localisation, validated protocols, controls and antibodies — the at-a-glance facts below, then the full design guide.
Must know before staining
| Expected localisation | Cytoplasmic staining in GI tract, bladder, prostate and CNS (HPA tissue IHC) | |
| Staining pattern | Goblet/glandular cytoplasm; cerebellar granular cytoplasm/membrane (HPA tissue IHC) | |
| Antigen retrieval | Citrate pH 6.0 HIER, 95–98 °C, 20 min (rule: cytoplasmic / membrane antigen) | |
| Positive control | Appendix+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 paraffin sections. (standard IHC practice; not target-specific) | |
| Caveat | GI positives vary: colon endocrine; small-intestinal goblet cells (HPA tissue IHC) | |
| Regulation | Down-regulated in some colorectal tumors (UniProt) | |
| Isoform / epitope | Two isoforms; no TM segment; check isoform epitope coverage (UniProt) |
Section 1
Recommended BCAS1 IHC & IF Protocols
The catalog antibody protocol is followed by one published chromogenic IHC protocol for BCAS1 in human multiple sclerosis brain lesions (PMC11772157).
| Sample | Tissue sections; selected-image fixative not specified (standard IHC workflow) |
| Fixation | Image fixative and duration unreported (datasheet A13589); 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 citrate buffer, pH 6.0, 20 min at 95–98 °C (standard rule: cytoplasmic / membrane 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 anti-BCAS1, 5 μg/mL (datasheet A13589) |
| 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 | BCAS1-positive staining in goblet cells of appendix (HPA tissue IHC: High). HPA tissue profile: Cytoplasmic expression in the gastrointestinal tract, urinary bladder, prostate and the CNS. No signal in the no-primary control. |
💡Decision noteStart with citrate pH 6.0 retrieval at 95–98 °C for 20 min (page antigen retrieval). The published protocol also reports Tris-EDTA pH 9.0 (PMC11772157).
Section 2
What Is the Expected BCAS1 Staining Pattern?
BCAS1 should chiefly appear in the cytoplasm on IHC sections, with strong staining in selected epithelial populations, including prostate glandular cells and intestinal goblet cells (HPA: tissue IHC, High). The protein is cytoplasmic and has no transmembrane segment (UniProt O75363: localisation and topology). HPA rates its tissue staining “Enhanced,” meaning consistency with RNA expression while external verification remains pending (HPA: reliability description).
| Distinct cytoplasmic reaction product in prostate glandular cells or duodenal goblet cells, with limited staining in adjacent cells. | This supports the expected cell-selective pattern: both populations are rated High by tissue IHC (HPA: prostate and duodenum). Assess the stained cell type as well as intensity; HPA reports cytoplasmic expression across several tissues, but its cell-level findings do not imply that every cell in a positive section should stain (HPA: tissue IHC profile). |
| Predominantly nuclear staining, or a broad extracellular deposit, instead of a cell-associated cytoplasmic pattern. | Treat this as discordant with the reported localisation (UniProt O75363: Cytoplasm; HPA: tissue IHC profile). General IHC practice: inspect the counterstain and a negative control before assigning the signal to BCAS1; review detection and wash conditions if the same pattern appears across unrelated structures. |
| Strong staining in adipocytes while expected epithelial cells show little or none. | HPA reports BCAS1 as Not detected in adipose-tissue adipocytes, whereas prostate glandular and duodenal goblet cells are High (HPA: tissue IHC). General IHC practice: consider antibody cross-reactivity or endogenous detection activity; compare a suitable negative control and check whether the signal follows cell morphology. |
| Haze or granular colour spread across the section, obscuring cell borders and the counterstain. | That distribution prevents a reliable call on the selective cytoplasmic pattern reported for BCAS1 (HPA: tissue IHC profile; UniProt O75363: Cytoplasm). General IHC practice: examine reagent-only or primary-omission controls, blocking, washes and development time to distinguish nonspecific background from interpretable cell-associated signal. |
| No convincing signal in the expected cells of a prostate or duodenum section. | An absent result conflicts with HPA's High calls for prostate glandular cells and duodenal goblet cells (HPA: tissue IHC). General IHC practice: first confirm that the relevant cells are present, then check the positive control, primary antibody conditions, detection reagents and antigen retrieval before interpreting the specimen as negative. |
💡Expected BCAS1 appearanceCall a positive result when a clear, cell-associated cytoplasmic signal is strong in prostate glandular or intestinal goblet cells (HPA: High; UniProt O75363: Cytoplasm); widespread nuclear, extracellular or adipocyte staining is a warning pattern (HPA: tissue IHC).
| Choice of reference tissue and cells | Prostate glandular cells, duodenal goblet cells and small-intestine goblet cells are High, while adipose-tissue adipocytes are Not detected (HPA: tissue IHC). Compare the named cell populations, since a whole-section average can conceal the reported distribution. |
| Strength and scope of validation | The tissue profile is rated Enhanced for consistency between staining and RNA expression, with external verification pending (HPA: reliability description). HPA lists Enhanced IHC status for HPA051816, HPA054745 and CAB033558 (HPA: antibody validation); those ratings do not validate an unlisted antibody. |
| Isoform and epitope interpretation | Two BCAS1 isoforms are recorded, but their antibody epitopes are not supplied here (UniProt O75363: isoforms; HPA: supplied antibody data). Do not infer which isoform an observed stain represents or use an apparent difference in staining to assign isoform-specific expression. |
| Does IF/ICC show the same pattern? | HPA approves a vesicular ICC-IF location, while UniProt gives the broader cytoplasmic location (HPA: subcellular ICC-IF; UniProt O75363: Cytoplasm). The vesicular observation concerns ICC-IF images and should not be imposed as the required appearance of chromogenic tissue IHC. |
| Situation | Likely cause | Next action |
|---|---|---|
| Positive tissue is blank. | The expected cells may be absent from the examined area, or an IHC workflow step may have failed; prostate glandular and duodenal goblet cells are High in HPA tissue IHC (HPA: tissue IHC). | Locate the named cells on the counterstained section. As general IHC practice, check a positive control, reagent preparation, retrieval conditions, primary incubation and detection before scoring the specimen. |
| Signal is weak in a tissue expected to be positive. | HPA's High rating applies to specific cells, not every part of that tissue (HPA: tissue IHC). Weak signal may also reflect general workflow conditions; no BCAS1-specific fixation effect is reported in the supplied sources. | Reassess the named cell population. As general IHC practice, compare the positive control and review validated retrieval, antibody incubation and chromogen development conditions. |
| Adipocytes stain strongly. | That finding conflicts with HPA's Not detected call for adipose-tissue adipocytes (HPA: tissue IHC); nonspecific binding or endogenous detection activity is possible under general IHC practice. | Check a primary-omission control and the detection system; reassess blocking and cell identification. Preserve the cell-specific distinction: the HPA breast entry also concerns adipocytes, not every breast cell (HPA: tissue IHC). |
| Signal is mainly nuclear. | Nuclear dominance is inconsistent with the recorded cytoplasmic localisation (UniProt O75363: Cytoplasm; HPA: tissue IHC profile). | Use the counterstain to verify compartment assignment. As general IHC practice, compare control sections and inspect background, antibody conditions and detection before accepting nuclear signal as specific. |
| Diffuse colour masks cell boundaries. | The distribution cannot be matched confidently to HPA's cell-specific cytoplasmic observations (HPA: tissue IHC); general IHC causes include excess background or overdevelopment. | As general IHC practice, compare the negative control, review blocking and washes, and shorten chromogen development where appropriate; then rescore only clearly localised cellular staining. |
| CNS staining seems inconsistent across cell types. | HPA reports High cerebellar granular-cell staining but Low hippocampal and caudate glial-cell staining; UniProt reports high brain expression, particularly in oligodendrocytes (HPA: tissue IHC; UniProt O75363: tissue specificity). | Record region and cell identity before comparing results. Do not convert UniProt's broader brain-expression statement into an expectation of uniformly strong IHC staining in all glial cells. |
Experiment evidence
IHC & IF images showing BCAS1 staining
A13589 · IHCImmunohistochemistry of BCAS1 in human breast carcinoma with BCAS1 antibody at 5 μg/mL.
A13589 · IFImmunofluorescence of BCAS1 in Human Breast Carcinoma cells with BCAS1 antibody at 20 μg/mL.
HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000064787-BCAS1/tissue/cerebral+cortex#imid_17754779
HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000064787-BCAS1/tissue/colon#imid_17754713
HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000064787-BCAS1/tissue/liver#imid_17754714
HPA · KidneyHuman Protein Atlas IHC, Kidney. Source: https://www.proteinatlas.org/ENSG00000064787-BCAS1/tissue/kidney#imid_17754715
HPA · ICC-IF MCF-7Human Protein Atlas immunofluorescence, MCF-7, antibody HPA054745; green = BCAS1, blue = nucleus, red = microtubules.
HPA · ICC-IF MCF-7Human Protein Atlas immunofluorescence, MCF-7, antibody HPA054745; green = BCAS1, blue = nucleus, red = microtubules.
← swipe / scroll for more images →
Real IHC / IF data behind the staining-pattern expectations in this section.
Sample controls for BCAS1 IHC & IF
🧪Run appendix first: goblet cells should stain strongly (HPA: High in appendix goblet cells). Use adipose tissue as the negative: adipocytes are not detected (HPA: Not detected in adipocytes); on the appendix slide, use only independently verified BCAS1-negative nongoblet cells as internal negatives and expect background-level chromogen (standard IHC practice).
Positive control tissue: Appendix (Goblet cells, HPA High)
Negative control tissue: Adipose tissue (HPA Not detected)
ICC-IF cell lines (HPA subcellular resource): HPA ICC-IF images show BCAS1 in MCF-7, RT-4, U2OS, with annotated localisation: Vesicles (approved) (HPA subcellular).
Technical controls: Include a no-primary, secondary-only control and a host-species-matched nonimmune IgG control, matched to the primary antibody’s isotype when applicable (standard IHC practice). Confirm specificity with BCAS1 knockout material or peptide blocking when a suitable immunizing peptide is available, and quench endogenous peroxidase for chromogenic IHC (standard IHC practice).
⚠️Feasibility: A target-specific fixation window, fixation effect, and retrieval dependency are unreported in the supplied evidence; the selected A13589 breast carcinoma IHC caption also leaves the fixative unreported (A13589 tissue-IHC caption). There is no matched comparison showing that frozen sections or IF are easier; vesicular ICC-IF localization is reported, but does not establish section-processing conditions (HPA: Vesicles, approved). Goblet-cell mucin can complicate interpretation of appendix staining, so compare the signal with the no-primary control and cell morphology (standard IHC practice).
HPA tissue IHC evidence for BCAS1
Comprehensive Human Protein Atlas IHC scoring per tissue (reliability: Enhanced — High consistency between antibody staining and RNA expression data. Pending external verification.). Rows are taken directly from the HPA tissue chart — click any row's HPA link to view the source.
Positive expression · recommended positive controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| Appendix | Goblet cells | High | Protein (IHC) | HPA → |
| Bronchus | Respiratory epithelial cells | High | Protein (IHC) | HPA → |
| Cerebellum | Granular cells - cytoplasm/membrane | High | Protein (IHC) | HPA → |
| Colon | Endocrine cells | High | Protein (IHC) | HPA → |
| Duodenum | Goblet cells | High | 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 → |
| Bone marrow | Hematopoietic cells | Not detected | Protein (IHC) | HPA → |
| Breast | Adipocytes | Not detected | Protein (IHC) | HPA → |
| Cervix | Glandular cells | Not detected | Protein (IHC) | HPA → |
Section 3
Advanced BCAS1 IHC Tips
Troubleshoot BCAS1 chromogenic IHC in paraffin sections by checking retrieval, compartment, cell identity, controls, and scoring before interpreting staining.
How should I adjust retrieval when BCAS1 staining is weak or patchy?
Start paraffin-section chromogenic IHC with citrate pH 6.0 HIER at 95–98 °C for 20 min (page retrieval setting). Cool slides in buffer, then compare staining with a matched positive control while keeping antibody concentration and detection conditions constant (standard IHC practice). If staining remains weak, test a higher-pH retrieval buffer on adjacent sections as a fallback, watching for tissue damage and increased background (standard IHC practice). Score the expected cytoplasmic signal separately from diffuse staining because BCAS1 is cytoplasmic and lacks a transmembrane segment (UniProt O75363 subcellular location and topology). Record retrieval conditions for every comparison (standard IHC practice).
Could fixation explain inconsistent BCAS1 staining across paraffin blocks?
Target-specific BCAS1 sensitivity to fixation is unknown from the supplied evidence, so do not assign a preferred fixative or fixation duration (supplied evidence). The carcinoma image caption gives 5 μg/mL antibody but does not state its fixative (A13589 tissue-IHC caption). Document each block’s fixative and processing history, and compare sections stained in the same run with identical retrieval and detection settings (standard IHC practice). Use a consistently stained control section to distinguish a block-dependent loss of signal from a run-wide failure (standard IHC practice). Assess morphology before calling faint cytoplasmic staining negative (standard IHC practice).
Where should a credible BCAS1 IHC signal appear?
Expect predominantly cytoplasmic staining: UniProt places BCAS1 in the cytoplasm, and the tissue-IHC profile reports cytoplasmic expression (UniProt O75363 subcellular location; HPA tissue IHC profile). Punctate cytoplasmic staining can be examined alongside this pattern because approved IF localisation is vesicular (HPA subcellular). Compare the signal within identified cell populations; HPA reports high staining in prostate glandular cells and cerebellar granular cells, among others (HPA tissue IHC). Do not score isolated nuclear colour as established BCAS1 localisation without independent validation (UniProt O75363 subcellular location; standard IHC practice). Use morphology and a counterstain to separate cellular signal from pigment or precipitate (standard IHC practice).
How can isoforms or epitope placement affect my IHC result?
BCAS1 has 2 annotated isoforms, so check whether the antibody’s documented immunogen covers the isoform your question requires (UniProt O75363 isoforms; standard IHC practice). The supplied caption does not identify an epitope; its 5 μg/mL image concentration cannot establish isoform coverage (A13589 tissue-IHC caption). BCAS1 also has annotated phosphoserines, including residues 124, 192, and 314, but their effect on this antibody’s staining is unknown (UniProt O75363 modified residues; supplied evidence). If results disagree across samples, compare an independently validated antibody with a documented epitope and retain identical processing controls (standard IHC practice). Do not infer isoform abundance from staining intensity alone (standard IHC practice).
How can IF help check BCAS1 cell identity and localisation?
Use IF as a companion localisation experiment and multiplex BCAS1 with a validated oligodendrocyte-lineage marker when examining brain, where oligodendrocyte expression is reported (UniProt O75363 tissue specificity; standard IF practice). Include a nuclear counterstain and inspect single channels before accepting apparent overlap (standard IF practice). Choose a red or far-red fluorophore after checking tissue autofluorescence and detector bleed-through (standard IF practice). Because BCAS1 is cytoplasmic with no transmembrane segment, permeabilise sufficiently to access an intracellular epitope, then optimise against morphology and background (UniProt O75363 subcellular location and topology; standard IF practice). Its approved vesicular IF localisation offers a further pattern check (HPA subcellular).
What should I change when BCAS1 DAB staining is diffuse?
First inspect a no-primary control for nonspecific detection and endogenous colour, and compare it with a positive-control section from the same run (standard IHC practice). Block endogenous peroxidase before chromogenic detection and titrate the IHC-validated antibody if widespread DAB obscures cellular boundaries (standard IHC practice). The carcinoma caption documents 5 μg/mL for its image, which is a reference point rather than a universal working concentration (A13589 tissue-IHC caption). Shorten chromogen development or improve washing when precipitate or diffuse deposition dominates (standard IHC practice). Judge residual signal against BCAS1’s reported cytoplasmic distribution and cell-specific tissue staining (UniProt O75363 subcellular location; HPA tissue IHC).
How should I quantify heterogeneous BCAS1 staining in tissue sections? ⚠ ANSWER MARKED FOR VERIFICATION
Define the cell population and viable tissue area before scoring, then apply the same threshold and sampling scheme across slides (standard IHC practice). For cellular cytoplasmic staining, report the percentage of positive cells and an H-score using intensity 0–3 multiplied by percentages, yielding 0–300 (standard IHC practice). If counting discrete positive cells, report density per mm² of viable, annotated tissue and the number of cells assessed (standard IHC practice). Normalise comparisons to the same cell type or compartment, since HPA staining varies among named cell populations (HPA tissue IHC). Exclude necrosis, folds, and section edges using prespecified rules (standard IHC practice).
How do I distinguish true BCAS1 staining from artefact?
Prioritise cytoplasmic signal within identifiable cells, consistent with BCAS1 localisation and the tissue-IHC profile (UniProt O75363 subcellular location; HPA tissue IHC profile). A prostate glandular-cell section offers a documented high-staining comparison, while adipocytes are listed as not detected (HPA tissue IHC). Treat nuclear-only colour, sharp section-edge gradients, and staining confined to necrosis as suspect until controls and morphology support them (UniProt O75363 subcellular location; standard IHC practice). Check no-primary and peroxidase-block controls when DAB appears in cells or regions inconsistent with the expected pattern (standard IHC practice). Interpret the breast-carcinoma image as observed staining at 5 μg/mL, without assuming an unreported fixative (A13589 tissue-IHC caption).
Boster reagents
Best BCAS1 / Breast carcinoma-amplified sequence 1 IHC Antibodies
The IHC-validated anti-BCAS1 antibody has human breast carcinoma tissue IHC data (A13589 IHC image caption) and human breast carcinoma cell IF data (A13589 IF image caption).
Real IHC data
Anti-BCAS1 Antibody
Cat # A13589
A13589 is listed for human IHC-P and IF (catalog: applications and reactivity). Its figures show IHC in human breast carcinoma at 5 μg/mL and IF in human breast carcinoma cells at 20 μg/mL (A13589 image captions).
Which to pick: Choose A13589 for paraffin-section IHC (catalog: IHC-P); its IHC figure shows human breast carcinoma, but the fixative is unreported (A13589 IHC image caption). For IF/ICC planning, A13589 has IF data in human breast carcinoma cells; ICC is not separately listed (A13589 IF image caption; catalog: applications). No cross-species option is supported because A13589 lists human reactivity only (catalog: reactivity).
Each figure is that product's own IHC / IF validation image from its datasheet.
References
- UniProt Consortium. UniProt entry O75363 (BCAS1_HUMAN, Breast carcinoma-amplified sequence 1).
- Human Protein Atlas. BCAS1 tissue IHC expression (reliability: Enhanced).
- Human Protein Atlas. BCAS1 subcellular location (ICC-IF): Localized to vesicles..
- Human Protein Atlas. BCAS1 antibody validation summary (3 antibodies).
- BCAS1 defines a heterogeneous cell population in diffuse gliomas. Oncotarget 2024 — PMC10812236.
- Inflammatory microglia correlate with impaired oligodendrocyte maturation in multiple sclerosis. Frontiers in immunology 2024 — PMC11772157.
- BCAS1 expression defines a population of early myelinating oligodendrocytes in multiple sclerosis lesions. Science translational medicine 2017 — PMC7116798.
- Postmortem quantitative MRI disentangles histological lesion types in multiple sclerosis. Brain pathology (Zurich, Switzerland) 2023 — PMC10580009.
- PubMed PMID:9671742 — UniProt-cited evidence.
- PubMed PMID:11780052 — UniProt-cited evidence.
- PubMed PMID:15489334 — UniProt-cited evidence.
Immunohistochemistry protocol sheet for BCAS1 (Breast carcinoma-amplified sequence 1). Expected localisation Cytoplasmic staining in GI tract, bladder, prostate and CNS (HPA tissue IHC). Antigen retrieval: Citrate pH 6.0 HIER, 95–98 °C, 20 min (rule: cytoplasmic / membrane antigen). Validated IHC antibody: A13589 at 5 μg/mL (the product's own IHC validation image is shown). Controls (HPA): positive Appendix (Goblet cells); negative Adipose tissue. IHC workflow; specimen preparation unspecified. Protocol steps: 1. Sample: Tissue sections; selected-image fixative not specified (standard IHC workflow); 2. Fixation: Image fixative and duration unreported (datasheet A13589); 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 citrate buffer, pH 6.0 , 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 anti-BCAS1, 5 μg/mL (datasheet A13589); 9. Primary incubation: Overnight at 4 °C (standard). Watch for: Fixation: Keep fixation consistent across paraffin sections. (standard IHC practice; not target-specific); Caveat: GI positives vary: colon endocrine; small-intestinal goblet cells (HPA tissue IHC); Isoform / epitope: Two isoforms; no TM segment; check isoform epitope coverage (UniProt); Regulation: Down-regulated in some colorectal tumors (UniProt). Evidence: UniProt O75363, HPA tissue IHC · Enhanced, HPA ICC-IF · vesicles., Datasheet A13589, PMC11772157. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/BCAS1/immunohistochemistry-ihc-guide.