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- Table of Contents
Plan chromogenic BMP1 IHC in paraffin sections using 1:50 as a starting dilution for the catalog antibody (datasheet: 1:50). Use colon glandular cells as a positive control (HPA tissue IHC), and account for BMP1 secretion when interpreting staining (UniProt).
Expected localisation, validated protocols, controls and antibodies — the at-a-glance facts below, then the full design guide.
| Expected localisation | Cytoplasmic staining in most tissues (HPA tissue IHC) | |
| Staining pattern | High glandular cell cytoplasmic staining in colon and duodenum (HPA tissue IHC) | |
| Antigen retrieval | Citrate pH 6.0 HIER, 95–98 °C, 20 min (rule: cytoplasmic / membrane antigen) | |
| Positive control | Colon+4 more · see all | |
| Negative control | Adipose tissue+4 more · see all |
| Fixation | Keep fixation consistent across paraffin sections. (standard IHC practice; not target-specific) | |
| Caveat | Secreted BMP1 may stain away from expressing cells (HPA tissue IHC) | |
| Regulation | Regulation unreported in the supplied record (UniProt) | |
| Isoform / epitope | 7 isoforms; propeptide epitopes may miss mature BMP1 (UniProt) |
The catalog antibody’s IHC-P protocol (datasheet) appears alongside three published BMP1 IHC methods (PMC5326928; PMC8971496; PMC6635762).
| Sample | Paraffin-embedded human skin tissue; fixative not specified (datasheet A02436) |
| Fixation | Image fixative and duration unreported (datasheet A02436); 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-BMP1, 1:50 recommended; image 1:200 (datasheet A02436) |
| 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 | BMP1-positive staining in glandular cells of colon (HPA tissue IHC: High). HPA tissue profile: Cytoplasmic expression in most tissues. No signal in the no-primary control. |
BMP1 is a secreted protein associated with the Golgi, extracellular space and extracellular matrix; it has no transmembrane segment (UniProt P13497: location, topology). In tissue IHC, expect mainly cytoplasmic staining in many cell types, especially glandular cells of the colon, duodenum and small intestine, cardiomyocytes, and placental trophoblastic cells (HPA tissue IHC: High). HPA rates the tissue profile Enhanced, while noting medium consistency with RNA because secreted protein can accumulate away from its source (HPA tissue IHC).
| Clear cytoplasmic staining in intestinal glandular cells, cardiomyocytes or placental trophoblastic cells. | This matches reported High staining in those cell types (HPA tissue IHC). Extracellular staining can also be plausible because BMP1 is secreted and associates with matrix (UniProt P13497: location). Judge the named cells and their compartment, rather than assigning one intensity to the entire tissue. |
| A sharply nuclear-only or membrane-rim pattern dominates the positive cells. | Neither pattern is the expected tissue profile: HPA reports predominantly cytoplasmic staining, and UniProt lists secreted and Golgi locations with no transmembrane segment (HPA tissue IHC; UniProt P13497: location, topology). Treat the pattern as suspect until staining controls and antibody specificity are checked. |
| Strong staining appears in adipocytes or squamous epithelial cells used as comparison cells. | Adipocytes in adipose tissue and breast, and squamous epithelial cells in cervix, esophagus and oral mucosa, are reported Not detected (HPA tissue IHC). Unexpected signal suggests cross-reactivity or detection background; a positive cell elsewhere in the section does not change those cell-specific comparisons. |
| Pale color covers most compartments, including tissue spaces and comparison cells. | A uniform haze is less persuasive than identifiable cytoplasmic staining in the reported positive cells (HPA tissue IHC: profile, High cells). Consider nonspecific detection or substrate background, and assess a no-primary control under the same imaging and development conditions (standard IHC practice). |
| No staining is visible in colon or duodenal glandular cells. | Both are reported High and are useful positive comparisons (HPA tissue IHC). An empty result could reflect a failed detection run or an antibody and retrieval combination that has not been shown to work; HPA supplies no BMP1-specific fixation or retrieval effect. |
| Secretion and compartment | BMP1 occurs in the trans-Golgi network and outside cells, including matrix, and lacks a transmembrane segment (UniProt P13497: location, topology). Thus cytoplasmic and extracellular signals can represent different pools; HPA documents the tissue IHC pattern mainly as cytoplasmic (HPA tissue IHC). |
| Processing and matrix deposition | The precursor has a signal peptide at residues 1–22 and propeptide at 23–120; the listed mature chain begins at 121 (UniProt P13497: processing). Interaction with POSTN promotes matrix deposition (UniProt P13497: subunit). The antibody epitope is unspecified here, so these facts cannot predict which processed form it detects. |
| Isoforms and antibody validation | Seven BMP1 isoforms are listed (UniProt P13497: isoforms). The supplied record does not map the IHC antibody's epitope across them. HPA014572 is listed as IHC Enhanced (HPA antibodies), supporting the reported tissue pattern without establishing equal detection of every isoform or extracellular pool. |
| Tissue and RNA comparison | HPA reports low tissue RNA specificity but cell-specific IHC differences, from High intestinal glandular staining to Not detected adipocytes (HPA tissue IHC). Because BMP1 is secreted, HPA cautions that RNA and protein locations may differ; score observed protein by cell and compartment rather than using RNA alone. |
| Situation | Likely cause | Next action |
|---|---|---|
| No signal in colon or duodenal glandular cells. | These cells are reported High, so a blank positive comparison raises concern about the staining run (HPA tissue IHC). The record does not identify a BMP1-specific retrieval requirement. | Check the catalog antibody's IHC-P instructions, primary dilution and detection reagents; compare a known-positive section processed in the same run (standard IHC practice). Do not infer a BMP1 fixation effect from this result. |
| Signal appears in HPA Not detected comparison cells. | Staining of adipocytes or listed squamous epithelial cells conflicts with their cell-specific HPA calls (HPA tissue IHC). Cross-reactivity or chromogenic background is possible. | Check cell identity and the no-primary control; reassess the IHC-validated antibody and detection settings before calling those cells BMP1-positive (standard IHC practice). |
| Predominantly nuclear or sharply membrane-rim staining. | The pattern does not fit HPA's cytoplasmic tissue profile or UniProt's secreted, Golgi and matrix locations and lack of a transmembrane segment (HPA tissue IHC; UniProt P13497: location, topology). | Compare reported positive and Not detected cell types in the same run, then check antibody specificity and secondary-only staining (HPA tissue IHC; standard IHC practice). |
| Diffuse brown haze obscures cell boundaries. | Uniform background prevents the cell-specific assessment needed for the HPA profile (HPA tissue IHC). Nonspecific binding or endogenous chromogenic detection activity are general IHC possibilities (standard IHC practice). | Inspect no-primary and reagent controls; review blocking, washes, detection time and counterstain so identifiable cells remain readable (standard IHC practice). |
| Cytoplasmic staining is present, but matrix signal is absent. | HPA reports predominantly cytoplasmic tissue staining, while extracellular and matrix locations come from UniProt; neither source requires visible matrix staining in every section (HPA tissue IHC; UniProt P13497: location). | Score the supported cytoplasmic pattern first. Report extracellular staining separately, and avoid treating its absence alone as a failed IHC run. |
| IF/ICC Q&A: should a punctate fluorescence pattern be called BMP1-positive? | HPA lists BMP1 as secreted but provides no ICC-IF images or main cellular location; the supplied antibody has no ICC validation entry (HPA subcellular; HPA antibodies). | Treat the pattern as unconfirmed. Use appropriate fluorescence controls and independently validate cell identity and compartment before interpreting it (standard IF practice); consult the separate IF/ICC guide for that application. |
Comprehensive Human Protein Atlas IHC scoring per tissue (reliability: Enhanced — Medium consistency between antibody staining and RNA expression data. Secreted protein, tissue location of RNA and protein is expected to differ.). 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 |
|---|---|---|---|---|
| Colon | Glandular cells | High | Protein (IHC) | HPA → |
| Duodenum | Glandular cells | High | Protein (IHC) | HPA → |
| Heart muscle | Cardiomyocytes | High | Protein (IHC) | HPA → |
| Placenta | Trophoblastic cells | High | Protein (IHC) | HPA → |
| Small intestine | Glandular cells | High | Protein (IHC) | HPA → |
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| Adipose tissue | Adipocytes | Not detected | Protein (IHC) | HPA → |
| Breast | Adipocytes | Not detected | Protein (IHC) | HPA → |
| Cervix | Squamous epithelial cells | Not detected | Protein (IHC) | HPA → |
| Esophagus | Squamous epithelial cells | Not detected | Protein (IHC) | HPA → |
| Liver | Cholangiocytes | Not detected | Protein (IHC) | HPA → |
Troubleshoot BMP1 staining in paraffin sections by checking retrieval, compartment, controls and scoring before interpreting chromogenic signal.
A02436 has real IHC data in paraffin-embedded human skin (A02436 image caption). IF is listed without a figure (A02436 applications/IF image alts); human, mouse, and rat reactivity is listed (A02436 reactivity).
A02436 will render with an IHC image of paraffin-embedded human skin at 1:200 (A02436 image caption). Its catalog lists IHC and IF applications and human, mouse, and rat reactivity, but supplies no IF image (A02436 applications/reactivity/IF image alts).
Which to pick: For tissue IHC, choose A02436: its own image documents paraffin-embedded human skin at 1:200, with the fixative unreported (A02436 image caption). For IF/ICC planning, A02436 lists IF at 1:50, but supplies no IF figure or explicit ICC application (A02436 applications/IF dilution/IF image alts). For cross-species work, A02436 is a rabbit polyclonal with human, mouse, and rat reactivity listed; its supplied tissue image shows human skin only (A02436 host/dilution_raw/reactivity/image caption).