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BCAM / Basal cell adhesion molecule · IHC design guide
Design Immunohistochemistry for BCAM
Plan BCAM chromogenic IHC on paraffin sections, expecting membranous staining at basal membranes and endothelial cells (HPA tissue IHC). Start with the catalog antibody's 0.5–1 μg/mL IHC range (datasheet A03148-1), and use kidney Bowman's capsule as a positive reference (HPA tissue IHC).
Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
BCAM 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 | Membranous at basal membranes and endothelium (HPA tissue IHC) | |
| Staining pattern | Membranous staining in basal membranes and endothelial cells (HPA tissue IHC) | |
| Antigen retrieval | EDTA pH 8.0 HIER, heat-mediated (datasheet A03148-1) | |
| Positive control | Cervix+4 more · see all | |
| Negative control | None in HPA (detected in all 45 tissues); use no-primary + isotype controls |
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 | Staining and RNA show medium consistency (HPA tissue IHC) | |
| Regulation | Expression regulation not specified (UniProt) | |
| Isoform / epitope | No isoforms listed; extracellular versus cytoplasmic epitope matters (UniProt) |
Section 1
Recommended BCAM IHC & IF Protocols
The catalog antibody’s IHC-P protocol is paired with published BCAM staining protocols for prostate, ovarian and renal cancer samples (PMC12599034; PMC9842900; PMC9361577).
| Sample | Paraffin-embedded human thyroid cancer tissue; fixative not specified (datasheet A03148-1) |
| Fixation | Image fixative and duration unreported (datasheet A03148-1); 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 A03148-1); 20 min, 95–100 °C (standard) |
| Peroxidase block | 3% H2O2, 10 min, room temperature (standard) |
| Blocking | 10% goat serum (datasheet A03148-1) |
| Primary antibody | Rabbit anti-BCAM, 0.5-1μg/ml (datasheet A03148-1) |
| Primary incubation | Overnight at 4 °C (datasheet A03148-1) |
| Detection | Streptavidin-biotin complex (SABC), DAB chromogen (datasheet A03148-1) |
| Counterstain | Hematoxylin, blue, dehydrate and mount (standard) |
| Expected result | BCAM-positive staining in glandular cells of cervix (HPA tissue IHC: High). HPA tissue profile: Membranous expression in basal membranes and endothelial cells. No signal in the no-primary control. |
💡Decision noteStart with heat-mediated EDTA pH 8.0 retrieval for the catalog antibody (datasheet: A03148-1); the ovarian study used Trilogy for its different BCAM antibody (PMC9842900).
Section 2
What Is the Expected BCAM Staining Pattern?
BCAM is a cell-membrane protein with an extracellular region at residues 32–547 and one transmembrane segment at 548–568 (UniProt P50895 topology). In paraffin-section IHC, expect membranous staining along epithelial basal regions and in endothelial cells (HPA tissue IHC: Enhanced; medium consistency with RNA). HPA also reports high staining in selected epithelial and kidney structures, so judge the pattern by both compartment and cell type (HPA tissue IHC).
| Crisp membranous staining follows epithelial basal regions and vessel endothelium. | This matches the reported tissue profile and cell-membrane assignment (HPA tissue IHC; UniProt P50895 subcellular location). Judge each structure against its local morphology; a brown outline alone does not identify the cell type (general IHC practice). |
| Strong staining appears in Bowman's capsule, nasopharyngeal basal cells, or cervical glandular cells. | These are reported high-staining examples (HPA tissue IHC: High in each). High signal is also reported in fallopian-tube non-ciliated cells, skin sebaceous glands, and bladder urothelial cells (HPA tissue IHC). Intensity can vary by structure; compare like cells within the section (general IHC practice). |
| Predominantly nuclear or broadly cytoplasmic staining replaces the expected membrane pattern. | For tissue IHC, this conflicts with the reported membranous profile (HPA tissue IHC; UniProt P50895 subcellular location). Treat it as suspect and inspect morphology and controls before calling BCAM positive (general IHC practice). HPA's approved nucleolar ICC-IF location is a separate assay observation (HPA subcellular ICC-IF). |
| Strong staining dominates a cell population outside the reported tissue pattern. | Unexpected cell-type staining can reflect cross-reactivity or endogenous detection activity (general IHC practice). It needs independent confirmation before assignment to BCAM, especially because HPA notes stained cells or structures that were not annotated (HPA tissue IHC: reliability description). |
| The whole section looks hazy, or a known high-staining structure has no detectable signal. | Uniform haze obscures compartment and cell boundaries; absent signal in a reported high-staining structure makes a negative call uninterpretable until controls pass (general IHC practice; HPA tissue IHC: High examples). Assess these as separate background and assay-performance problems (general IHC practice). |
💡Expected BCAM appearanceCall a positive result when distinct membrane staining marks the relevant basal epithelial, endothelial, or listed high-staining cells (HPA tissue IHC; UniProt P50895 subcellular location); diffuse haze or dominant nuclear staining is a suspect IHC pattern (general IHC practice).
| Membrane topology and unknown antibody epitope | BCAM has an extracellular region, one transmembrane segment, and a cytoplasmic tail (UniProt P50895 topology). The supplied record does not locate the antibody epitope, so topology cannot justify a specific retrieval or permeabilisation setting (UniProt P50895 topology; general IHC practice). |
| Tissue and cell-type variation | HPA grades several epithelial or kidney structures High, breast myoepithelial and bronchial basal cells Medium, and selected neural or endocrine cells Low (HPA tissue IHC). Use the named cell population when comparing intensity; the tissue name alone is too broad (general IHC practice). |
| Strength and limits of tissue evidence | The tissue IHC profile is Enhanced, with medium consistency between antibody staining and RNA; some stained structures are unannotated (HPA tissue IHC: reliability description). The listed antibody, HPA005654, has Enhanced IHC validation (HPA antibodies). Interpret unexpected structures cautiously. |
| IHC versus ICC-IF localisation | HPA reports an approved nucleoli fibrillar-center location in ICC-IF, while its tissue IHC profile is membranous (HPA subcellular ICC-IF; HPA tissue IHC). If IF/ICC is the intended assay, use its separate guide; the ICC observation does not redefine a tissue-IHC positive result. |
| Situation | Likely cause | Next action |
|---|---|---|
| Known high-staining tissue is blank. | A failed staining run, inadequate detection, or an unsuitable assay setting is possible (general IHC practice); the supplied sources establish no BCAM-specific fixation or retrieval sensitivity. | Check the same-run positive control, antibody and detection steps, and retrieval conditions used for the validated IHC assay (general IHC practice). Do not score the specimen negative until the positive control works. |
| Brown signal fills nuclei or cytoplasm without clear cell borders. | The result does not match the tissue membranous profile (HPA tissue IHC; UniProt P50895 subcellular location); nonspecific staining is possible (general IHC practice). | Inspect a matched negative detection control and tissue morphology, then reassess blocking, antibody concentration, and detection conditions (general IHC practice). Require a convincing membrane pattern before assigning tissue-IHC positivity. |
| Vessels or unrelated cell populations look positive in a confusing pattern. | Endothelium is a reported BCAM-positive compartment (HPA tissue IHC); signal elsewhere could include endogenous detection activity or cross-reactivity (general IHC practice). | Identify vessel lining on the counterstained section, compare a negative detection control, and seek independent antibody or orthogonal support for unexpected cells (general IHC practice; HPA tissue IHC: unannotated staining caveat). |
| The slide has diffuse brown background across multiple compartments. | Excess detection signal or insufficient blocking may obscure localisation (general IHC practice); HPA does not identify a BCAM-specific cause (HPA tissue IHC). | Compare a negative detection control, then adjust blocking, antibody concentration, washes, or chromogen development as appropriate to the assay (general IHC practice). Reassess only after membranes can be distinguished. |
| A low-staining cell type gives little or no signal. | Low staining is reported for Purkinje, hippocampal neuronal, caudate glial, and selected endocrine glandular cells (HPA tissue IHC: Low); absence there alone does not establish assay failure. | Check a reported high-staining structure on a same-run control before interpreting the low-staining sample (HPA tissue IHC: High examples; general IHC practice). Record the specific cell type scored. |
| ICC-IF appears nucleolar while paraffin-section IHC appears membranous. | Those are the respective HPA assay observations (HPA subcellular ICC-IF: approved nucleoli fibrillar center; HPA tissue IHC: membranous profile). | Interpret each image within its assay and controls (general IHC practice). Use the separate IF/ICC guide for that application; do not use the nucleolar ICC-IF observation as the expected paraffin-section IHC pattern. |
Experiment evidence
IHC & IF images showing BCAM staining
A03148-1 · IHCIHC analysis of CD239/BCAM using anti-CD239/BCAM antibody (A03148-1). CD239/BCAM was detected in paraffin-embedded section of human thyroid
A03148-1 · IHCIHC analysis of BCAM using anti-BCAM antibody (A03148-1). BCAM was detected in a paraffin-embedded section of human kidney tissue. Heat medi
A03148-1 · IFIF analysis of CD239/BCAM using anti-CD239/BCAM antibody (A03148-1). CD239/BCAM was detected in immunocytochemical section of Hela cells. En
HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000187244-BCAM/tissue/cerebral+cortex#imid_6577288
HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000187244-BCAM/tissue/colon#imid_6577214
HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000187244-BCAM/tissue/liver#imid_6577215
HPA · KidneyHuman Protein Atlas IHC, Kidney. Source: https://www.proteinatlas.org/ENSG00000187244-BCAM/tissue/kidney#imid_6577208
HPA · ICC-IF A-431Human Protein Atlas immunofluorescence, A-431, antibody HPA005654; green = BCAM, blue = nucleus, red = microtubules.
HPA · ICC-IF A-431Human Protein Atlas immunofluorescence, A-431, antibody HPA005654; green = BCAM, blue = nucleus, red = microtubules.
← swipe / scroll for more images →
Real IHC / IF data behind the staining-pattern expectations in this section.
Sample controls for BCAM IHC & IF
🧪Run skin first and score sebaceous glands for strong staining (HPA: High in skin sebaceous glands). HPA lists no negative tissue because BCAM is detected in all 45 scored tissues; use no-primary and isotype controls, and score any cells without specific staining on the positive slide as internal background comparators without assuming a named cell type is BCAM-negative (HPA: no negative rows; standard IHC practice).
Positive control tissue: Cervix (Glandular cells, HPA High)
Negative control tissue: None in HPA: BCAM is detected in all 45 scored tissues. Use a no-primary (secondary-only) and an isotype control instead.
ICC-IF cell lines (HPA subcellular resource): HPA ICC-IF images show BCAM in A-431, U-251MG, U2OS, with annotated localisation: Nucleoli fibrillar center (approved) (HPA subcellular).
Technical controls: Include a secondary-only section, a concentration-matched rabbit isotype or nonimmune IgG control appropriate to the antibody’s clonality, and BCAM-knockout tissue or peptide competition if the immunizing peptide is available (caption: rabbit primary; standard IHC practice). For the caption’s SABC–DAB detection, quench endogenous peroxidase and assess endogenous biotin background; distinguish skin pigment from DAB signal (caption: SABC–DAB; standard IHC practice).
⚠️Feasibility: A target-specific fixation window and fixation effect are unreported, and the selected A03148-1 paraffin-section caption does not state a fixative (caption: fixative unreported). That caption supports heat retrieval in EDTA at pH 8.0 and primary antibody at 1 μg/ml overnight at 4°C for paraffin IHC; retrieval dependence beyond this example is unreported (caption: A03148-1 tissue IHC). No supplied comparison establishes frozen sections or IF as easier, and skin pigment may complicate DAB scoring (supplied evidence: no frozen-section or comparative IF protocol; standard IHC practice).
HPA tissue IHC evidence for BCAM
Comprehensive Human Protein Atlas IHC scoring per tissue (reliability: Enhanced — Medium consistency between antibody staining and RNA expression data. Antibody staining in cells/structures not annotated, view images.). 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 |
|---|---|---|---|---|
| Cervix | Glandular cells | High | Protein (IHC) | HPA → |
| Fallopian tube | Non-ciliated cells | High | Protein (IHC) | HPA → |
| Kidney | Bowman's capsule | High | Protein (IHC) | HPA → |
| Nasopharynx | Basal cells | High | Protein (IHC) | HPA → |
| Skin | Sebaceous glands | High | Protein (IHC) | HPA → |
Undetected expression · recommended negative controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| None in HPA: BCAM is detected in all 45 scored tissues. Use a no-primary (secondary-only) and an isotype control instead. | ||||
Section 3
Advanced BCAM IHC Tips
For BCAM chromogenic IHC, assess membrane staining in the appropriate cell compartment and compare it with the documented tissue pattern (UniProt P50895; HPA tissue IHC).
What retrieval should I start with if BCAM staining is weak in paraffin sections?
Start with heat-mediated retrieval in EDTA at pH 8.0 for paraffin sections (datasheet A03148-1). The selected BCAM image used that retrieval before overnight incubation at 4°C with 1 μg/mL primary antibody (datasheet A03148-1). If staining remains weak, compare a modestly longer heating interval with the initial run, keeping section thickness, antibody concentration and detection constant (standard IHC practice). Check for membranous signal in basal epithelial or endothelial cells while watching for diffuse background or tissue damage (HPA tissue IHC; standard IHC practice). Change one retrieval variable at a time so any improvement can be attributed to that change (standard IHC practice).
How should I troubleshoot fixation when BCAM membrane staining is inconsistent?
The selected paraffin-section caption does not state a fixative, so BCAM-specific sensitivity to fixation is unknown (datasheet A03148-1). Record the fixative, fixation duration, processing schedule and section age for each specimen before comparing staining intensity (standard IHC practice). Run sections with different fixation histories in the same staining batch, using the same EDTA pH 8.0 retrieval and detection conditions (datasheet A03148-1; standard IHC practice). Compare membrane staining in the relevant cell populations and examine morphology for processing damage (HPA tissue IHC; standard IHC practice). Do not infer a BCAM-specific fixation effect from its membrane topology or tissue distribution alone (UniProt P50895 topology; HPA tissue IHC).
Which staining compartment should count as convincing BCAM signal?
Prioritise staining at cell surfaces, especially along basal epithelial membranes and in endothelial cells (UniProt P50895 subcellular location; HPA tissue IHC). BCAM has an extracellular region at residues 32–547, a transmembrane segment at 548–568 and a cytoplasmic tail at 569–628 (UniProt P50895 topology). Use morphology and a counterstain to distinguish a cell border from adjacent extracellular material in the chromogenic section (standard IHC practice). HPA also reports an approved nucleolar location in its subcellular dataset, creating a compartment discrepancy to investigate (HPA subcellular; HPA tissue IHC). Score nuclear signal separately until its specificity is resolved with appropriate controls (standard IHC practice).
Could epitope location explain different BCAM staining patterns between antibodies?
Yes: accessibility and staining can depend on whether an antibody recognises the extracellular region or cytoplasmic tail (UniProt P50895 topology; standard IHC practice). BCAM spans the membrane at residues 548–568, leaving residues 32–547 extracellular and 569–628 cytoplasmic (UniProt P50895 topology). The record lists 0 isoforms, but that entry does not establish the epitope of the catalog antibody (UniProt P50895 isoforms; datasheet A03148-1). Five annotated glycosylation sites lie in the extracellular region, so document any epitope information supplied for a comparison antibody before interpreting differences (UniProt P50895 glycosylation and topology; standard IHC practice). Compare antibodies on matched sections with their own validated retrieval conditions (standard IHC practice).
How can I use IF to examine a disputed BCAM IHC pattern?
Use IF as a separate localisation check and multiplex BCAM with a validated marker for the expected basal epithelial or endothelial cell population (HPA tissue IHC; standard IF practice). Select fluorophores after checking tissue autofluorescence and place the weaker signal in a spectrally clean channel (standard IF practice). If the antibody binds the extracellular region, begin with minimal permeabilisation; access to a cytoplasmic epitope may require permeabilisation (UniProt P50895 topology; standard IF practice). The catalog IHC caption does not identify its epitope or establish an IF protocol, so optimise IF fixation and detergent conditions independently (datasheet A03148-1; standard IF practice). Include single-label and secondary-only controls to assess bleed-through and background (standard IF practice).
What should I check when BCAM DAB staining is diffuse or widespread?
First compare the suspect signal with cell-border staining in the expected basal epithelial and endothelial structures (HPA tissue IHC). The selected caption used 10% goat serum blocking, a biotinylated secondary antibody, a streptavidin–biotin detection complex and DAB (datasheet A03148-1). Include a no-primary control, check endogenous peroxidase blocking and assess whether the biotin-based detection contributes background (standard IHC practice; datasheet A03148-1). Titrate the primary antibody around the documented 1 μg/mL condition while holding retrieval and development time constant (datasheet A03148-1; standard IHC practice). Evaluate section edges and damaged tissue separately before assigning diffuse colour to BCAM (standard IHC practice).
How should I score BCAM staining across sections with different cell composition? ⚠ ANSWER MARKED FOR VERIFICATION
Define the cell population and membrane-positive criterion before scoring, then report the percentage of positive cells and membrane intensity or an H-score (HPA tissue IHC; standard IHC practice). For vascular analysis, report positive vessels per mm² alongside total vessel density or the fraction of vessels positive for BCAM (HPA tissue IHC; standard IHC practice). For epithelial analysis, normalise positive counts to the number of evaluable cells in the specified compartment (standard IHC practice). Keep retrieval, DAB development, imaging and thresholds consistent across sections (standard IHC practice). Report scores for basal epithelial, endothelial and other compartments separately because BCAM expression is not restricted to one tissue population (UniProt P50895 tissue specificity; HPA tissue IHC).
How can I distinguish true BCAM positivity from chromogenic artefact?
A convincing result follows cell membranes in plausible basal epithelial or endothelial populations and remains visible away from section edges (UniProt P50895 subcellular location; HPA tissue IHC; standard IHC practice). High staining has also been reported in kidney Bowman’s capsule and nasopharyngeal basal cells, providing tissue-level comparisons when those structures are present (HPA tissue IHC). Treat isolated nuclear colour cautiously because the HPA subcellular dataset reports a nucleolar location while its tissue IHC profile is membranous (HPA subcellular; HPA tissue IHC). Exclude necrotic regions, edge staining and colour reproduced in a no-primary control from the positive score (standard IHC practice). Check endogenous peroxidase and biotin-related background when interpreting the caption’s DAB and biotin-based workflow (datasheet A03148-1; standard IHC practice).
Boster reagents
Best BCAM / Basal cell adhesion molecule IHC Antibodies
A03148-1 has IHC images from human paraffin sections and an IF image from HeLa cells (catalog image captions); its listed reactivity includes human, mouse and rat (catalog applications and reactivity).
Real IHC data
Anti-CD239/BCAM Antibody ®
Cat # A03148-1
A03148-1 will render with its human thyroid cancer IHC figure; its catalog also shows human kidney IHC and HeLa IF images (catalog image captions). M03148 is listed for IHC in human, mouse and rat, but has no supplied IHC or IF image and will not render a card (catalog applications, reactivity and image fields).
Which to pick: Choose A03148-1 for paraffin-section IHC: its own captions document staining in human thyroid cancer and kidney sections, with the fixative unreported (A03148-1 IHC image captions). Choose A03148-1 for IF/ICC because its HeLa image documents IF staining; M03148 has no listed IF application (catalog IF image caption and applications). Both list human, mouse and rat reactivity, but the supplied tissue IHC images document human samples only; M03148 is monoclonal, while A03148-1 has no listed clone (catalog reactivity, clone fields and IHC image captions).
Each figure is that product's own IHC / IF validation image from its datasheet.
References
- UniProt Consortium. UniProt entry P50895 (BCAM_HUMAN, Basal cell adhesion molecule).
- Human Protein Atlas. BCAM tissue IHC expression (reliability: Enhanced).
- Human Protein Atlas. BCAM subcellular location (ICC-IF): Localized to the nucleoli fibrillar center..
- Human Protein Atlas. BCAM antibody validation summary (1 antibodies).
- Unveiling prognostic biomarkers and immunotherapeutic insights in prostate cancer through multi-omics and machine learning. European journal of medical research 2025 — PMC12599034.
- Single-Cell Sequencing Unveils the Heterogeneity of Nonimmune Cells in Chronic Apical Periodontitis. Frontiers in cell and developmental biology 2021 — PMC8883837.
- Basal cell adhesion molecule promotes metastasis-associated processes in ovarian cancer. Clinical and translational medicine 2023 — PMC9842900.
- Integrated multi-omics analyses reveal that BCAM is associated with epigenetic modification and tumor microenvironment subtypes of clear cell renal cell carcinoma. Clinical epigenetics 2022 — PMC9361577.
- PubMed PMID:7777537 — UniProt-cited evidence.
- PubMed PMID:15057824 — UniProt-cited evidence.
- PubMed PMID:15489334 — UniProt-cited evidence.
Immunohistochemistry protocol sheet for BCAM (Basal cell adhesion molecule). Expected localisation Membranous at basal membranes and endothelium (HPA tissue IHC). Antigen retrieval: EDTA pH 8.0 HIER, heat-mediated (datasheet A03148-1). Validated IHC antibody: A03148-1 at 0.5-1μg/ml (the product's own IHC validation image is shown). Controls (HPA): positive Cervix (Glandular cells); negative None in HPA (detected in all 45 tissues); use no-primary + isotype controls. IHC-P workflow on paraffin sections; fixative unspecified. Protocol steps: 1. Sample: Paraffin-embedded human thyroid cancer tissue; fixative not specified (datasheet A03148-1); 2. Fixation: Image fixative and duration unreported (datasheet A03148-1); 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 A03148-1); 20 min, 95–100 °C (standard); 6. Peroxidase block: 3% H2O2, 10 min , room temperature (standard); 7. Blocking: 10% goat serum (datasheet A03148-1); 8. Primary antibody: Rabbit anti-BCAM, 0.5-1μg/ml (datasheet A03148-1); 9. Primary incubation: Overnight at 4 °C (datasheet A03148-1). Watch for: Fixation: Keep fixation consistent across paraffin sections. (standard IHC practice; not target-specific); Caveat: Staining and RNA show medium consistency (HPA tissue IHC); Isoform / epitope: No isoforms listed; extracellular versus cytoplasmic epitope matters (UniProt); Regulation: Expression regulation not specified (UniProt). Evidence: UniProt P50895, HPA tissue IHC · Enhanced, HPA ICC-IF · nucleoli fibrillar center., Datasheet A03148-1, PMC12599034, PMC9842900. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/BCAM/immunohistochemistry-ihc-guide.