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- Table of Contents
This guide uses cytoplasmic tissue staining (HPA tissue IHC) and ER membrane localisation (UniProt) to plan BCAP31 paraffin IHC. It pairs a catalog antibody starting range of 2–5 μg/mL (datasheet A03767-4) with high-staining lung macrophages and unstained cardiomyocytes as reference populations (HPA tissue IHC).
Expected localisation, validated protocols, controls and antibodies — the at-a-glance facts below, then the full design guide.
| Expected localisation | Tissue: cytoplasm (HPA tissue IHC); protein: ER membrane (UniProt) | |
| Staining pattern | Widespread cytoplasmic staining; high in lung macrophages (HPA tissue IHC) | |
| Antigen retrieval | EDTA pH 8.0 HIER, heat-mediated (datasheet A03767-4) | |
| Positive control | Cerebellum+4 more · see all | |
| Negative control | Heart muscle+4 more · see all |
| Fixation | Keep fixation consistent across sections (standard IHC practice; not target-specific) | |
| Caveat | Presumed off-target binding may confound staining (HPA tissue IHC) | |
| Regulation | ER stress redistributes BCAP31 (UniProt) | |
| Isoform / epitope | 2 isoforms; epitope differences unspecified (UniProt) |
The catalog antibody’s IHC-P protocol is followed by 3 published BCAP31 IHC protocols with usable staining details (PMC11683684; PMC6771250; PMC7055246).
| Sample | Paraffin-embedded human colon cancer tissue; fixative not specified (datasheet A03767-4) |
| Fixation | Image fixative and duration unreported (datasheet A03767-4); 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 A03767-4); 20 min, 95–100 °C (standard) |
| Peroxidase block | 3% H2O2, 10 min, room temperature (standard) |
| Blocking | 10% goat serum (datasheet A03767-4) |
| Primary antibody | Rabbit anti-BCAP31, 2-5 μg/ml (datasheet A03767-4) |
| Primary incubation | Overnight at 4 °C (datasheet A03767-4) |
| Detection | HRP-conjugated secondary, DAB chromogen (datasheet A03767-4) |
| Counterstain | Hematoxylin, blue, dehydrate and mount (standard) |
| Expected result | BCAP31-positive staining in bergmann glia - cytoplasm/membrane of cerebellum (HPA tissue IHC: High). HPA tissue profile: Ubiquitous cytoplasmic expression. No signal in the no-primary control. |
BCAP31 is an endoplasmic reticulum (ER) membrane protein with three transmembrane segments (UniProt P51572 topology). In paraffin-section IHC, expect mainly cytoplasmic or membrane-associated staining, particularly in cerebellar Bergmann glia and lung macrophages (HPA: High in both). Expression is widespread, but intensity varies by cell type (HPA: ubiquitous cytoplasmic profile). Treat the reference pattern as a guide: HPA rates its tissue IHC evidence Supported, with medium consistency and presumed off-target binding observed (HPA: tissue IHC reliability).
| Cytoplasmic/membrane staining in Bergmann glia or lung macrophages, with little nuclear signal. | This fits the reported High staining in those cells (HPA: tissue IHC) and BCAP31’s ER membrane location (UniProt P51572). Judge the pattern within the identified cells; section-wide color alone cannot establish specific staining (general IHC practice). |
| Predominantly nuclear staining, or a pattern confined to an unexpected compartment. | A nuclear-dominant result conflicts with the ER localisation reported by UniProt and ICC-IF (UniProt P51572; HPA: subcellular ER). Review morphology and controls before interpreting it as BCAP31; HPA reports presumed off-target binding in its tissue assessment (HPA: tissue IHC reliability). |
| Strong color in cells listed as Not detected, such as cardiomyocytes or skeletal myocytes. | That differs from the HPA reference for those specific cells (HPA: Not detected in heart and skeletal muscle). Consider cross-reactivity or endogenous detection activity, then check controls (general IHC practice). An HPA Not detected call is an observed staining result, not proof that every specimen must be negative. |
| Diffuse color across cells and surrounding tissue, without a discernible cytoplasmic pattern. | Treat this as possible background until controls support a cell-associated signal (general IHC practice). The reference is ubiquitous cytoplasmic expression, not uniform staining of every part of a section (HPA: tissue IHC profile); evaluate background separately from genuine low-level staining. |
| No discernible signal in Bergmann glia or lung macrophages. | This disagrees with the reported High IHC staining in those cells (HPA: tissue IHC). First confirm that the relevant cells are present and identifiable; then assess the staining run and antibody conditions (general IHC practice). A failed reference section cannot by itself establish BCAP31 absence in test tissue. |
| Compartment and topology | BCAP31 occupies the ER and ER–Golgi intermediate compartment membranes, with three transmembrane segments (UniProt P51572). A cytoplasmic or membrane-associated tissue pattern is therefore plausible; the record does not specify an IHC epitope or retrieval condition. |
| Cell-type variation | HPA reports High staining in Bergmann glia and lung macrophages, Medium in several listed cell types, and Not detected in certain others (HPA: tissue IHC). Compare like cells across sections; a low or negative reference call does not define every cell in an organ. |
| Antibody evidence | The three listed antibodies, HPA003906, CAB015350 and CAB015424, each have Supported IHC status (HPA: antibody validation). HPA also notes medium tissue IHC consistency and presumed off-target binding, so use staining distribution and controls together (HPA: tissue IHC reliability). |
| Isoforms and protein processing | Two isoforms are recorded, with no signal peptide or propeptide and no annotated glycosylation sites or modified residues (UniProt P51572). The supplied record gives no isoform-specific tissue pattern or antibody epitope; do not assign a staining difference to a particular isoform. |
| IF/ICC Q&A: What pattern can help assess localisation? | HPA reports enhanced ER localisation in ICC-IF, with images from A-431, U-251MG and U2OS (HPA: subcellular). That supports an ER-pattern comparison, while the tissue IHC profile remains the reference for this paraffin-section guide (HPA: tissue IHC). |
| Situation | Likely cause | Next action |
|---|---|---|
| The positive reference section is blank. | The run may have failed, the expected cells may be absent from the section, or the signal may be below detection (general IHC practice). | Confirm Bergmann glia or lung macrophages are present, then review the documented IHC-P staining conditions and run controls (HPA: High in those cells; general IHC practice). |
| Reference cells stain weakly despite a clear positive control. | Intensity varies across cell types and assay runs; HPA’s High category is an observed reference pattern, not a prescribed color threshold (HPA: tissue IHC). | Compare the same cell type under matched staining and counterstain conditions; record whether the cytoplasmic pattern remains discernible (general IHC practice). |
| Nuclei dominate the apparent signal. | This conflicts with reported ER localisation and may reflect nonspecific staining or interpretation of overlapping structures (UniProt P51572; HPA: subcellular ER; general IHC practice). | Inspect cell boundaries and the counterstain, compare control sections, and avoid calling nuclear color BCAP31 without independent support (general IHC practice). |
| Color appears in HPA Not detected cell types. | Cross-reactivity or endogenous detection activity can produce misleading chromogenic color (general IHC practice); HPA reports presumed off-target binding (HPA: tissue IHC reliability). | Check an appropriate negative reagent control and the detection-system controls; interpret against the exact cell type in the HPA reference (general IHC practice; HPA: tissue IHC). |
| The entire section has diffuse background. | Insufficient blocking, excess detection reagent, or incomplete removal of unbound reagent can obscure localisation (general IHC practice). | Review blocking, reagent exposure and washes in the documented IHC-P workflow; require identifiable cell-associated cytoplasmic staining before scoring (general IHC practice; HPA: cytoplasmic profile). |
| Two antibodies give different cell distributions. | HPA lists Supported IHC validation for each listed antibody but reports medium consistency and presumed off-target binding overall (HPA: antibody validation and tissue IHC reliability). | Compare the same cells in matched sections, check controls for each antibody, and describe the disagreement rather than assigning either pattern to BCAP31 automatically (general IHC practice). |
Comprehensive Human Protein Atlas IHC scoring per tissue (reliability: Supported — Medium consistency between antibody staining and RNA expression data. Presumed off target binding observed and disregarded.). Rows are taken directly from the HPA tissue chart — click any row's HPA link to view the source.
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| Cerebellum | Bergmann glia - cytoplasm/membrane | High | Protein (IHC) | HPA → |
| Lung | Macrophages | High | Protein (IHC) | HPA → |
| Adipose tissue | Adipocytes | Medium | Protein (IHC) | HPA → |
| Adrenal gland | Glandular cells | Medium | Protein (IHC) | HPA → |
| Appendix | Glandular cells | Medium | Protein (IHC) | HPA → |
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| Heart muscle | Cardiomyocytes | Not detected | Protein (IHC) | HPA → |
| Oral mucosa | Squamous epithelial cells | Not detected | Protein (IHC) | HPA → |
| Skeletal muscle | Myocytes | Not detected | Protein (IHC) | HPA → |
| Soft tissue | Peripheral nerve | Not detected | Protein (IHC) | HPA → |
| Vagina | Squamous epithelial cells | Not detected | Protein (IHC) | HPA → |
These questions focus on interpreting BCAP31 staining in paraffin sections, with one entry covering IF/ICC.
IHC images show BCAP31 staining in human colon cancer, rat lung and human brain sections (catalog IHC captions). IF images show A549 cells and human paraffin sections (A03767-4 IF captions).
A03767-4 shows IHC in human colon cancer paraffin sections and IF in A549 cells and human paraffin sections (A03767-4 image captions); A03767 shows IHC in formaldehyde-fixed rat lung paraffin sections (A03767 IHC caption). M03767 shows IHC in formaldehyde-fixed human brain paraffin sections (M03767 IHC caption).
Which to pick: For human tissue IHC, choose A03767-4 for paraffin sections at 2–5 μg/mL with EDTA pH 8.0 retrieval; its IHC caption does not report a fixative (A03767-4 datasheet and IHC caption). M03767 is a mouse monoclonal option for paraffin-section human brain paraffin sections at 1:25 with citrate pH 6 retrieval (M03767 catalog and IHC caption); for IF/ICC, choose A03767-4 at 5 μg/mL, supported by images of A549 cells and human paraffin sections (A03767-4 datasheet and IF captions). For cross-species planning, A03767 is a rabbit polyclonal reactive with human, mouse and rat, but its IHC image and stated IHC validation cover rat tissue only (A03767 catalog, dilution notes and IHC caption). The selected A03767-4 tissue-IHC caption documents paraffin sections, but does not specify the fixative (selected-SKU IHC image A03767-4).