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BAMBI / BMP and activin membrane-bound inhibitor homolog · IHC design guide
Design Immunohistochemistry for BAMBI
Plan BAMBI chromogenic IHC on paraffin sections using the reported cytoplasmic tissue pattern (HPA tissue IHC) and membrane topology (UniProt). A human lung IHC example uses 5 μg/mL (datasheet); interpret staining in light of low antibody–RNA agreement (HPA tissue IHC).
Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
BAMBI 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 tissue staining (HPA tissue IHC); membrane protein (UniProt) | |
| Staining pattern | Cytoplasmic staining across several tissue types (HPA tissue IHC) | |
| Antigen retrieval | Citrate pH 6.0 HIER, 95–98 °C, 20 min (rule: cytoplasmic / membrane antigen) | |
| Positive control | Adipose tissue+4 more · see all | |
| Negative control | Cervix+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 | Low agreement between antibody staining and RNA expression (HPA tissue IHC) | |
| Regulation | Expression varies by tissue (UniProt) | |
| Isoform / epitope | No isoforms annotated; extracellular and cytoplasmic regions (UniProt) |
Section 1
Recommended BAMBI IHC & IF Protocols
The catalog antibody’s IHC-P protocol (page catalog) is accompanied by four published BAMBI IHC protocols (PMC11504530; PMC5097819; PMC12194233; PMC2890648).
| Sample | Tissue sections; selected-image fixative not specified (standard IHC workflow) |
| Fixation | Image fixative and duration unreported (datasheet A07983); 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-BAMBI, 5 μg/mL (datasheet A07983) |
| 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 | BAMBI-positive staining in adipocytes of adipose tissue (HPA tissue IHC: Medium). HPA tissue profile: Cytoplasmic expression in several different tissue types. No signal in the no-primary control. |
💡Decision noteStart with citrate pH 6.0 HIER at 95–98 °C for 20 min (page retrieval rule); compare the published Tris-EDTA method if staining is weak (PMC11504530).
Section 2
What Is the Expected BAMBI Staining Pattern?
BAMBI is a membrane protein with an extracellular region, one transmembrane segment and a cytoplasmic tail (UniProt Q13145 topology). In tissue IHC, the reported pattern is chiefly cytoplasmic, with medium staining in lung macrophages, adipocytes and bronchial respiratory epithelial cells (HPA tissue IHC). Treat this as a provisional expectation: HPA rates the tissue antibody Approved but reports low agreement between staining and RNA data, pending external verification (HPA tissue IHC).
| Distinct cytoplasmic chromogen in lung macrophages, with nearby unstained areas remaining clear. | This fits the reported medium macrophage signal and chiefly cytoplasmic tissue pattern (HPA tissue IHC). Score the identified cells and their staining intensity, rather than calling the entire lung section positive. A sharp membrane rim is not required by the tissue images: UniProt describes BAMBI topology, while HPA describes its observed IHC appearance (UniProt Q13145 topology; HPA tissue IHC). |
| Predominantly nuclear staining, or an even stain across nuclei and cytoplasm in most cells. | That is a poor match for the chiefly cytoplasmic tissue pattern (HPA tissue IHC). Review morphology, counterstain and staining controls before assigning BAMBI positivity (general IHC practice). The additional nucleolar location reported by HPA comes from ICC-IF and does not, by itself, validate a nuclear-only chromogenic tissue pattern (HPA subcellular ICC-IF). |
| Strong staining in colon glandular cells while the comparison lung macrophages stain as expected. | HPA reports colon glandular cells as not detected and lung macrophages at medium intensity (HPA tissue IHC). Consider antibody cross-reactivity or endogenous chromogenic detection activity, then examine appropriate detection controls (general IHC practice). This contrast raises concern; it does not prove that every stained colon cell is a false positive, given HPA's stated low staining–RNA consistency (HPA tissue IHC). |
| Weak color spread over stroma, empty spaces and multiple cell types without clear cellular boundaries. | This is diffuse background, not a convincing cell-specific pattern (general IHC practice). Compare a control lacking primary antibody and inspect washing and blocking steps (general IHC practice). HPA's reported cytoplasmic staining in selected cells cannot establish the identity of uniform background across a section (HPA tissue IHC). |
| No discernible stain in lung macrophages or another HPA-reported medium-positive cell population. | A negative result in a reported positive population warrants a run check, although medium HPA staining is not a guarantee for every specimen (HPA tissue IHC). Check tissue identification, antibody and detection controls, and the catalog antibody's IHC-P instructions (general IHC practice). HPA's Approved rating carries a low-consistency caveat, so avoid concluding that BAMBI is biologically absent from this result alone (HPA tissue IHC). |
💡Expected BAMBI appearanceCall a positive when identifiable lung macrophages or bronchial respiratory epithelial cells show cell-associated, chiefly cytoplasmic chromogen of roughly medium intensity (HPA tissue IHC); widespread acellular color or dominant nuclear-only staining is suspect against that reference pattern (HPA tissue IHC; general IHC practice).
| Topology and epitope location | BAMBI has residues 21–152 outside the cell, 153–173 across the membrane and 174–260 in the cytoplasm (UniProt Q13145 topology). The supplied record does not identify this antibody's epitope, so topology alone cannot predict which staining or retrieval condition will expose it. |
| Processing and glycosylation | UniProt lists a signal peptide at residues 1–20, a mature chain spanning 21–260 and one glycosylation site at residue 87 (UniProt Q13145 processing and glycosylation). These annotations describe the protein; they do not establish tissue-specific shedding or a target-specific fixation effect. |
| Strength of the tissue reference | The IHC antibody is Approved, while HPA reports low agreement between antibody staining and RNA expression and says external verification is pending (HPA tissue IHC; HPA antibodies). Use its cell-level pattern as a comparison, and seek independent support before treating an unexpected distribution as established BAMBI biology (general IHC practice). |
| IF/ICC: should its puncta determine the IHC call? | No. ICC-IF reports vesicles as the main location, with additional nucleoli fibrillar center and lipid-droplet signals (HPA subcellular ICC-IF). Those observations can help interpret an IF image, but they do not replace the chiefly cytoplasmic tissue IHC reference (HPA tissue IHC). |
| Situation | Likely cause | Next action |
|---|---|---|
| Reported medium-positive cells show no chromogen. | The staining run or detection may have failed; HPA also notes limited agreement with RNA data (general IHC practice; HPA tissue IHC). | Confirm cell identity and inspect positive and detection controls; follow the catalog antibody's IHC-P instructions for the run (general IHC practice). Do not infer BAMBI-specific fixation sensitivity from this outcome. |
| A nuclear-only pattern dominates the section. | It conflicts with the chiefly cytoplasmic tissue IHC profile; ICC-IF's additional nucleolar signal is a separate observation (HPA tissue IHC; HPA subcellular ICC-IF). | Check counterstain and a control lacking primary antibody, then require independent support before assigning nuclear tissue staining to BAMBI (general IHC practice). |
| Colon glandular cells stain strongly. | Those cells are listed as not detected, so cross-reactivity or endogenous detection activity is possible (HPA tissue IHC; general IHC practice). | Compare with the same-run detection control and an HPA-reported positive population such as lung macrophages; assess the specific cells, not section-wide color (HPA tissue IHC; general IHC practice). |
| Color is diffuse or appears in cell-free regions. | Diffuse background can arise from nonspecific reagent binding or residual detection reagent (general IHC practice). | Inspect a control lacking primary antibody and review blocking, washing and development against the chosen chromogenic IHC workflow (general IHC practice). |
| A crisp membrane rim is weak, but cytoplasmic staining is visible. | UniProt assigns BAMBI a transmembrane segment, whereas HPA describes tissue staining as cytoplasmic (UniProt Q13145 topology; HPA tissue IHC). | Score cell-associated cytoplasmic signal in the expected cell population; do not reject it solely for lacking a membrane outline (HPA tissue IHC; general IHC practice). |
| Replicate sections give different positive-cell distributions. | HPA reports low consistency between antibody staining and RNA expression, so its Approved tissue profile needs careful comparison (HPA tissue IHC). | Recheck cell identification and same-run controls, document the differing distributions, and seek independent validation before making a biological claim (general IHC practice). |
Experiment evidence
IHC & IF images showing BAMBI staining
A07983 · IHCImmunohistochemistry of BAMBI in human lung tissue with BAMBI antibody at 5 μg/ml.
A07983 · IFImmunofluorescence of BAMBI in human lung tissue with BAMBI antibody at 20 μg/ml.
HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000095739-BAMBI/tissue/cerebral+cortex#imid_7510728
HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000095739-BAMBI/tissue/colon#imid_7510662
HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000095739-BAMBI/tissue/liver#imid_7510655
HPA · KidneyHuman Protein Atlas IHC, Kidney. Source: https://www.proteinatlas.org/ENSG00000095739-BAMBI/tissue/kidney#imid_7510664
HPA · ICC-IF MCF-7Human Protein Atlas immunofluorescence, MCF-7, antibody HPA010819; green = BAMBI, blue = nucleus, red = microtubules.
HPA · ICC-IF MCF-7Human Protein Atlas immunofluorescence, MCF-7, antibody HPA010819; green = BAMBI, blue = nucleus, red = microtubules.
← swipe / scroll for more images →
Real IHC / IF data behind the staining-pattern expectations in this section.
Sample controls for BAMBI IHC & IF
🧪Run adipose tissue first and assess staining in adipocytes (HPA: Medium in adipocytes). Use cervix glandular cells as the negative tissue (HPA: Not detected in cervix glandular cells); compare other cells on the adipose slide with background, but do not assume they are BAMBI-negative because their expression is unspecified (HPA: adipocyte-specific row).
Positive control tissue: Adipose tissue (Adipocytes, HPA Medium)
Negative control tissue: Cervix (HPA Not detected)
ICC-IF cell lines (HPA subcellular resource): HPA ICC-IF images show BAMBI in MCF-7, SK-MEL-30, U2OS, with annotated localisation: Vesicles (approved) (HPA subcellular).
Technical controls: Include a no-primary, secondary-only control and a host- and isotype-matched control for a monoclonal antibody, or species-matched nonimmune IgG for a polyclonal antibody (standard IHC practice). Use BAMBI knockout tissue as a biological specificity control if available, and check endogenous peroxidase background in adipose sections before chromogenic scoring (standard IHC practice).
⚠️Feasibility: A target-specific fixation window or fixation effect is unreported, and the selected tissue-IHC caption does not state its fixative (selected SKU A07983 caption). Antigen-retrieval dependence is unreported; the supplied evidence does not establish whether frozen sections or IF would be easier (supplied target/application evidence). Lipid extraction during paraffin processing can leave adipocyte vacuoles that complicate scoring (standard histology practice).
HPA tissue IHC evidence for BAMBI
Comprehensive Human Protein Atlas IHC scoring per tissue (reliability: Approved — Low 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 |
|---|---|---|---|---|
| Adipose tissue | Adipocytes | Medium | Protein (IHC) | HPA → |
| Bone marrow | Hematopoietic cells | Medium | Protein (IHC) | HPA → |
| Bronchus | Respiratory epithelial cells | Medium | Protein (IHC) | HPA → |
| Cerebral cortex | Neuropil | Medium | Protein (IHC) | HPA → |
| Fallopian tube | Glandular cells | Medium | Protein (IHC) | HPA → |
Undetected expression · recommended negative controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| Cervix | Glandular cells | Not detected | Protein (IHC) | HPA → |
| Colon | Glandular cells | Not detected | Protein (IHC) | HPA → |
| Duodenum | Glandular cells | Not detected | Protein (IHC) | HPA → |
| Esophagus | Squamous epithelial cells | Not detected | Protein (IHC) | HPA → |
| Oral mucosa | Squamous epithelial cells | Not detected | Protein (IHC) | HPA → |
Section 3
Advanced BAMBI IHC Tips
Troubleshoot BAMBI staining in paraffin sections by checking retrieval, cell type and compartment, then confirming patterns with appropriate controls.
How should I retrieve BAMBI in paraffin sections when staining is weak?
Use citrate buffer at pH 6.0 for heat-induced retrieval at 95–98 °C for 20 minutes (page antigen-retrieval method). Let sections cool in the buffer before washing, and process a known positive control alongside the test section to distinguish retrieval failure from low expression (standard IHC practice). BAMBI has an extracellular region at residues 21–152 and a cytoplasmic region at 174–260, so epitope accessibility can differ between antibodies (UniProt Q13145 topology). If staining remains weak, test a different retrieval buffer only as a controlled fallback, keeping antibody concentration and detection conditions constant (standard IHC practice).
Could fixation explain weak BAMBI staining in my paraffin sections?
BAMBI-specific sensitivity to fixation is unknown from the supplied evidence; the selected tissue caption gives an antibody concentration but does not state the fixative (A07983 tissue-IHC caption). Record the fixative, fixation duration, section age and retrieval conditions for each specimen before comparing staining intensity (standard IHC practice). Run sections from the same block through a small retrieval comparison while holding primary antibody and detection conditions constant (standard IHC practice). If staining varies between blocks, assess tissue preservation and control-antigen staining before attributing the difference to BAMBI expression (standard IHC practice). The lung caption used 5 µg/ml but provides no fixation comparison (A07983 tissue-IHC caption).
Should BAMBI appear at the cell membrane or in the cytoplasm?
BAMBI is annotated as a membrane protein with a transmembrane segment at residues 153–173 (UniProt Q13145 topology). Tissue IHC reports cytoplasmic staining across several tissue types, including medium staining in lung macrophages (HPA tissue IHC). Cell-based IF reports vesicles as the main location, with additional nucleolar fibrillar-center and lipid-droplet locations; these observations should be assessed separately from the tissue IHC pattern (HPA subcellular). At high magnification, score membrane-associated and cytoplasmic signal separately, and compare each with a no-primary control and tissue morphology (standard IHC practice). Treat diffuse staining across unrelated compartments as a reason to check background and antibody specificity (standard IHC practice).
How does BAMBI epitope position affect antibody choice and staining?
Check the antibody's stated immunogen or mapped epitope before interpreting a missing membrane signal, because the selected tissue caption does not identify an epitope (A07983 tissue-IHC caption; standard IHC practice). BAMBI has an extracellular region at residues 21–152, a transmembrane segment at 153–173 and a cytoplasmic region at 174–260 (UniProt Q13145 topology). Its signal peptide spans residues 1–20, and glycosylation is annotated at residue 87 (UniProt Q13145 processing and glycosylation). Compare antibodies against distinct mapped regions, if available, on adjacent sections under matched detection conditions (standard IHC practice). The supplied record lists 0 isoforms, so it provides no basis for isoform-specific staining claims (UniProt Q13145 isoforms).
How can I investigate BAMBI localisation by multiplex IF?
For IF, pair BAMBI with a macrophage marker such as CD68 when examining lung cells, because tissue IHC reports medium BAMBI staining in lung macrophages (HPA tissue IHC; standard IF practice). Select a far-red fluorophore for the weaker channel, and inspect an unstained section to identify tissue autofluorescence before assigning colors (standard IF practice). BAMBI's extracellular residues 21–152 may be accessible without permeabilisation, whereas an epitope within cytoplasmic residues 174–260 requires permeabilisation; confirm the antibody epitope first (UniProt Q13145 topology; standard IF practice). Compare matched permeabilised and nonpermeabilised preparations with single-color controls to assess colocalisation and bleed-through (standard IF practice).
What should I check when BAMBI IHC shows widespread brown staining?
Compare the section with a no-primary control and inspect whether staining follows tissue edges, necrotic areas or pigment rather than intact cell boundaries (standard IHC practice). Quench endogenous peroxidase before chromogenic detection, and use an appropriate protein block and thorough washes to limit nonspecific signal (standard IHC practice). Titrate the primary antibody downward from a tested starting condition if background remains high; the selected lung image used 5 µg/ml, which is evidence for that image rather than an optimum for every specimen (A07983 tissue-IHC caption). Interpret residual cytoplasmic staining cautiously because HPA reports low consistency between antibody staining and RNA expression (HPA tissue IHC reliability).
How should I quantify BAMBI staining across paraffin sections? ⚠ ANSWER MARKED FOR VERIFICATION
Define the cell population and compartment before scoring, then apply the same threshold, counterstain and image-acquisition settings to every section (standard IHC practice). Report the percentage of positive cells with an intensity-based H-score, or count positive cells per mm² when cell density is the outcome (standard IHC practice). Normalise counts to viable tissue area or to the number of eligible cells, and exclude folds, edges and necrotic regions using prespecified rules (standard IHC practice). Score lung macrophages separately from other lung cells when using that specimen, because HPA specifically reports medium staining in macrophages (HPA tissue IHC). Include control-slide performance and observer or analysis thresholds with the results (standard IHC practice).
When is a BAMBI-positive IHC pattern convincing rather than artefactual?
Give greatest weight to reproducible staining in intact cells that exceeds a no-primary control and remains consistent across serial sections (standard IHC practice). Check cell identity: HPA reports medium staining in lung macrophages, while its tissue profile describes cytoplasmic expression in several tissue types (HPA tissue IHC). BAMBI's membrane topology makes membrane-associated signal plausible, but a strong nuclear-only chromogenic pattern needs independent confirmation rather than assignment to BAMBI from topology alone (UniProt Q13145 topology; standard IHC practice). Exclude staining concentrated at cut edges, necrosis or sites of residual endogenous peroxidase before scoring (standard IHC practice). HPA flags low consistency between staining and RNA data, so corroborate unexpected patterns with another validated antibody or an independent assay (HPA tissue IHC reliability; standard IHC practice).
Boster reagents
Best BAMBI / BMP and activin membrane-bound inhibitor homolog IHC Antibodies
A07983 has IHC and IF images from human lung tissue (catalog image captions). Its listed reactivity covers human, mouse and rat (catalog reactivity).
Real IHC data
Anti-BAMBI Antibody
Cat # A07983
A07983 will render with an IHC image of human lung tissue at 5 μg/mL (IHC image caption). The same SKU has an IF image of human lung tissue at 20 μg/mL (IF image caption).
Which to pick: Choose A07983 for tissue IHC: IHC-P is listed, and its IHC image shows human lung tissue at 5 μg/mL (catalog applications; IHC image caption); the fixative is unreported (IHC image caption). Choose A07983 for IF because it has a human lung tissue IF image at 20 μg/mL; ICC validation is not listed (IF image caption; catalog applications). For cross-species work, A07983 lists human, mouse and rat reactivity, while its supplied IHC and IF images show human tissue (catalog reactivity; IHC and IF image captions).
Each figure is that product's own IHC / IF validation image from its datasheet.
References
- UniProt Consortium. UniProt entry Q13145 (BAMBI_HUMAN, BMP and activin membrane-bound inhibitor homolog).
- Human Protein Atlas. BAMBI tissue IHC expression (reliability: Approved).
- Human Protein Atlas. BAMBI subcellular location (ICC-IF): Mainly localized to vesicles. In addition localized to the nucleoli fibrillar center and lipid droplets..
- Human Protein Atlas. BAMBI antibody validation summary (2 antibodies).
- Hepatic Bone Morphogenetic Protein and Activin Membrane-Bound Inhibitor Levels Decline in Hepatitis C but Are Not Associated with Progression of Hepatocellular Carcinoma. Biomedicines 2024 — PMC11504530.
- Expression of BMP and Actin Membrane Bound Inhibitor Is Increased during Terminal Differentiation of MSCs. Stem cells international 2016 — PMC5097819.
- Comparison of Differentially Expressed Genes in Human and Canine Osteosarcoma. Life (Basel, Switzerland) 2025 — PMC12194233.
- The TGF-beta-pseudoreceptor BAMBI is strongly expressed in COPD lungs and regulated by nontypeable Haemophilus influenzae. Respiratory research 2010 — PMC2890648.
- PubMed PMID:8621228 — UniProt-cited evidence.
- PubMed PMID:15164054 — UniProt-cited evidence.
- PubMed PMID:15489334 — UniProt-cited evidence.
Immunohistochemistry protocol sheet for BAMBI (BMP and activin membrane-bound inhibitor homolog). Expected localisation Cytoplasmic tissue staining (HPA tissue IHC); membrane protein (UniProt). Antigen retrieval: Citrate pH 6.0 HIER, 95–98 °C, 20 min (rule: cytoplasmic / membrane antigen). Validated IHC antibody: A07983 at 5 μg/mL (the product's own IHC validation image is shown). Controls (HPA): positive Adipose tissue (Adipocytes); negative Cervix. 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 A07983); 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-BAMBI, 5 μg/mL (datasheet A07983); 9. Primary incubation: Overnight at 4 °C (standard). Watch for: Fixation: Keep fixation consistent across sections. (standard IHC practice; not target-specific); Caveat: Low agreement between antibody staining and RNA expression (HPA tissue IHC); Isoform / epitope: No isoforms annotated; extracellular and cytoplasmic regions (UniProt); Regulation: Expression varies by tissue (UniProt). Evidence: UniProt Q13145, HPA tissue IHC · Approved, HPA ICC-IF · Mainly localized to vesicles. In addition localized to the nucleoli…, Datasheet A07983, PMC11504530, PMC5097819. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/BAMBI/immunohistochemistry-ihc-guide.