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BMX / Cytoplasmic tyrosine-protein kinase BMX · IHC design guide
Design Immunohistochemistry for BMX
Plan chromogenic BMX IHC in paraffin sections using epididymal glandular cells as a positive reference (HPA tissue IHC). Start the IHC-validated antibody at 1:50–1:200 (datasheet), include a no-primary control (standard IHC practice), and interpret staining in light of reported off-target binding (HPA tissue IHC).
Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
BMX 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 | Cytoplasm in epididymal glandular cells (HPA tissue IHC) | |
| Staining pattern | Distinct cytoplasmic staining in epididymal glandular cells (HPA tissue IHC) | |
| Antigen retrieval | Citrate pH 6.0 HIER, 95–98 °C, 20 min (rule: cytoplasmic / membrane antigen) | |
| Positive control | Epididymis+1 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 | Presumed off-target staining; verification pending (HPA tissue IHC) | |
| Regulation | Highly expressed in migratory cells (UniProt) | |
| Isoform / epitope | No reported isoforms or processing; one cytoplasmic chain (UniProt) |
Section 1
Recommended BMX IHC & IF Protocols
The catalog antibody’s IHC-P protocol is accompanied by published BMX staining protocols for cervical cancer tissue and yak testicular tissue (PMC5564764; PMC10178277).
| Sample | Tissue sections; selected-image fixative not specified (standard IHC workflow) |
| Fixation | Image fixative and duration unreported (datasheet A02537-1); 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-BMX, 1:50-1:200 (datasheet A02537-1) |
| 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 | BMX-positive staining in glandular cells of epididymis (HPA tissue IHC: High). HPA tissue profile: Distinct cytoplasmic expression in epididymis. No signal in the no-primary control. |
💡Decision noteStart with citrate pH 6.0 HIER at 95–98 °C for 20 min (page antigen-retrieval rule); the cervical cancer study used a shorter pressure-cooker retrieval (PMC5564764).
Section 2
What Is the Expected BMX Staining Pattern?
In paraffin-section IHC, expect distinct cytoplasmic BMX staining in epididymal glandular cells, with High staining there and Medium staining in testicular Sertoli cells (HPA: tissue IHC). BMX is a cytoplasmic kinase with no transmembrane segment (UniProt P51813: localization and topology). HPA rates its IHC evidence Enhanced, while noting medium staining–RNA consistency, disregarded presumed off-target binding, and pending external verification (HPA: tissue IHC reliability).
| Distinct cytoplasmic staining in epididymal glandular cells; weaker staining in testicular Sertoli cells. | This matches the reported High epididymal and Medium Sertoli-cell patterns (HPA: tissue IHC). Judge compartment and cell identity together: HPA's Enhanced rating supports the observed IHC pattern, but its reliability note still calls for care when interpreting additional stained structures (HPA: tissue IHC reliability). |
| Predominantly nuclear or sharply membrane-only staining in the expected IHC-positive cells. | Recheck the chromogenic pattern before assigning it to BMX: the tissue IHC profile describes distinct cytoplasmic staining (HPA: tissue IHC). Plasma-membrane localization and additional nucleoplasmic localization come from ICC-IF, a different assay (HPA: subcellular ICC-IF); they do not establish either pattern as the expected paraffin-section IHC result. |
| Strong staining in adipocytes, bronchial respiratory epithelium, or bone-marrow hematopoietic cells. | Those sampled cell types are reported as Not detected by HPA tissue IHC (HPA: tissue IHC). Suspect cross-reactivity or endogenous chromogen-generating activity, then compare the same run's controls (general IHC practice). The HPA reliability note explicitly records presumed off-target binding that was disregarded (HPA: tissue IHC reliability). |
| Diffuse color across cells, stroma, and the negative-control section. | A field-wide deposit is difficult to reconcile with distinct cytoplasmic staining restricted to the reported positive cell types (HPA: tissue IHC). Background in the negative control points toward detection chemistry, insufficient blocking, or inadequate washing rather than a cell-specific BMX pattern (general IHC practice). |
| No convincing signal in epididymal glandular cells. | A negative result conflicts with their reported High staining (HPA: tissue IHC). Check tissue identity, positive-control performance, retrieval, antibody incubation, and chromogen development before interpreting the sample as BMX-negative (general IHC practice). HPA's Enhanced rating describes its antibody evidence; it does not guarantee every local staining run (HPA: antibodies; general IHC practice). |
💡Expected BMX appearanceCall a positive IHC result when epididymal glandular cells show distinct, predominantly cytoplasmic High staining; strong staining in HPA Not-detected cell types or dominant nuclear staining needs investigation (HPA: tissue IHC).
| Assay-specific compartment evidence | UniProt places BMX in the cytoplasm and notes cell-edge localization when complexed with BCAR1 (UniProt P51813: subcellular location). HPA tissue IHC reports cytoplasmic epididymal staining, whereas its ICC-IF record supports plasma-membrane localization with additional nucleoplasmic localization (HPA: tissue IHC; HPA: subcellular ICC-IF). Interpret each assay against its own evidence. |
| IHC antibody evidence | HPA lists 2 rabbit polyclonal antibodies with Enhanced IHC status, HPA001048 and CAB032495 (HPA: antibodies). Enhanced supports the reported IHC pattern, while the tissue summary reports only medium staining–RNA consistency, disregarded presumed off-target binding, and pending external verification (HPA: tissue IHC reliability). |
| Protein architecture and modification | BMX has no transmembrane segment, no listed signal peptide or propeptide, one chain spanning residues 1–675, and no listed glycosylation sites (UniProt P51813: topology, processing, glycosylation). UniProt also lists phosphotyrosines at 216, 224, and 566 (UniProt P51813: modified residues). Without an antibody epitope, these facts cannot predict retrieval needs or phospho-specific staining. |
| Choice of comparison tissue | Epididymal glandular cells provide the reported High pattern; testicular Sertoli cells provide a Medium comparison (HPA: tissue IHC). HPA reports Not detected staining in specified adipocytes, bronchial respiratory epithelial cells, and bone-marrow hematopoietic cells (HPA: tissue IHC). Compare the named cell types rather than treating every cell in an organ as a control. |
| Situation | Likely cause | Next action |
|---|---|---|
| Epididymal glandular cells are blank. | This misses HPA's reported High positive population (HPA: tissue IHC); a failed staining run remains possible (general IHC practice). | Verify tissue identity and examine a positive control from the same run; review retrieval, primary incubation, and detection steps (general IHC practice). No BMX-specific fixation sensitivity or retrieval condition is supplied. |
| Color appears throughout the section, including the negative control. | The distribution differs from HPA's distinct cytoplasmic pattern (HPA: tissue IHC); excess detection activity or nonspecific background is possible (general IHC practice). | Inspect reagent-only controls, endogenous enzyme blocking, primary concentration, and washing according to the detection system (general IHC practice). Do not score diffuse color as cell-specific BMX. |
| A HPA Not-detected cell population stains strongly. | The result conflicts with that population's reported tissue IHC level (HPA: tissue IHC); cross-reactivity or endogenous detection activity may contribute (general IHC practice). | Confirm the cell type, compare control sections, and review the staining distribution; HPA notes disregarded presumed off-target binding in its reliability summary (HPA: tissue IHC reliability; general IHC practice). |
| Most signal looks nuclear or confined to cell outlines. | That does not match the reported cytoplasmic paraffin-section pattern (HPA: tissue IHC). The separate ICC-IF record reports plasma-membrane and additional nucleoplasmic localization (HPA: subcellular ICC-IF). | Check morphology, counterstain, and controls before scoring the IHC slide; keep ICC-IF compartment observations separate when explaining the result (general IHC practice; HPA: subcellular ICC-IF). |
| Sertoli cells stain less intensely than epididymal glandular cells. | That ranking is consistent with Medium versus High staining in the respective reported cell types (HPA: tissue IHC). | Compare like cell populations and exposure or development conditions across sections before calling the weaker signal a failure (general IHC practice). |
| Can an IF/ICC image set the expected paraffin IHC pattern? | HPA's ICC-IF entry supports a main plasma-membrane location and an additional nucleoplasmic location, while its tissue IHC entry reports distinct cytoplasmic staining (HPA: subcellular ICC-IF; HPA: tissue IHC). | Use the cytoplasmic tissue IHC profile to assess chromogenic paraffin sections; consult the separate IF/ICC guide for fluorescence interpretation (HPA: tissue IHC; HPA: subcellular ICC-IF). |
Experiment evidence
IHC & IF images showing BMX staining
A02537-1 · IHCImmunohistochemistry (IHC) analyzes of ETK (K35) pAb in paraffin-embeddedhuman liver carcinoma tissue at 1:50.showing cytoplasmic and cell m
A02537-1 · IHCImmunohistochemistry (IHC) analyzes of ETK (K35) pAb in paraffin-embeddedhuman rectum carcinoma tissue at 1:50.showing cytoplasmic and cell
A02537Y43 · IHCImmunohistochemistry (IHC) analyzes of ETK (Y43) pAb in paraffin-embedded human tonsil carcinoma tissue at 1:50.
HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000102010-BMX/tissue/cerebral+cortex#imid_335582
HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000102010-BMX/tissue/colon#imid_335516
HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000102010-BMX/tissue/liver#imid_335517
HPA · KidneyHuman Protein Atlas IHC, Kidney. Source: https://www.proteinatlas.org/ENSG00000102010-BMX/tissue/kidney#imid_335510
HPA · ICC-IF HUVEC/TERT2Human Protein Atlas immunofluorescence, HUVEC/TERT2, antibody HPA001048; green = BMX, blue = nucleus, red = microtubules.
HPA · ICC-IF HUVEC/TERT2Human Protein Atlas immunofluorescence, HUVEC/TERT2, antibody HPA001048; green = BMX, blue = nucleus, red = microtubules.
← swipe / scroll for more images →
Real IHC / IF data behind the staining-pattern expectations in this section.
Sample controls for BMX IHC & IF
🧪Run epididymis first; its glandular cells should stain (HPA: High in epididymal glandular cells). Use adipose tissue adipocytes as a negative comparator (HPA: Not detected in adipocytes); putative internal negative cells on the epididymis slide should show only background chromogen, although the supplied HPA rows identify no validated BMX-negative cell type within epididymis (HPA: epididymis row).
Positive control tissue: Epididymis (Glandular cells, HPA High)
Negative control tissue: Adipose tissue (HPA Not detected)
ICC-IF cell lines (HPA subcellular resource): HPA ICC-IF images show BMX in HUVEC/TERT2, with annotated localisation: Plasma membrane (supported) (HPA subcellular).
Technical controls: Include no-primary/secondary-only and host- and Ig-class-matched nonimmune rabbit IgG controls for the rabbit polyclonal primary, plus a BMX knockout specimen if available (selected A02537-1 caption: pAb and anti-rabbit secondary; standard IHC practice). Quench endogenous peroxidase and check endogenous biotin background in the epididymis section when using avidin-peroxidase detection (selected A02537-1 caption: avidin-peroxidase; standard IHC practice).
⚠️Feasibility: The selected A02537-1 caption documents staining in a paraffin-embedded section at 1:50, but reports no fixative; a BMX-specific fixation window or fixation effect is unreported (selected A02537-1 caption). Antigen-retrieval conditions and retrieval dependency are unreported in the supplied evidence (selected A02537-1 caption; supplied target/application evidence). The HUVEC/TERT2 ICC-IF images do not establish whether frozen sections or tissue IF are easier, and no epididymis-specific artefact is reported (HPA: HUVEC/TERT2 ICC-IF; supplied target/application evidence).
HPA tissue IHC evidence for BMX
Comprehensive Human Protein Atlas IHC scoring per tissue (reliability: Enhanced — Medium consistency between antibody staining and RNA expression data. Presumed off target binding observed and disregarded. 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 |
|---|---|---|---|---|
| Epididymis | Glandular cells | High | Protein (IHC) | HPA → |
| Testis | Sertoli cells | Medium | 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 → |
| Appendix | Glandular cells | Not detected | Protein (IHC) | HPA → |
| Bone marrow | Hematopoietic cells | Not detected | Protein (IHC) | HPA → |
| Breast | Adipocytes | Not detected | Protein (IHC) | HPA → |
Section 3
Advanced BMX IHC Tips
Troubleshoot BMX staining in paraffin sections by checking retrieval, controls and cell compartment before scoring chromogenic signal.
How should I retrieve BMX in paraffin sections if staining is weak?
Start with citrate buffer at pH 6.0, heated to 95–98 °C for 20 minutes (page retrieval rule). Keep section thickness, cooling and detection conditions matched while comparing retrieval runs, so a change in signal can be attributed to the retrieval step (standard IHC practice). If staining remains weak, test a stronger retrieval condition on adjacent sections as a fallback, while checking for tissue damage and increased background (standard IHC practice). Compare the result with a positive control such as epididymal glandular cells (HPA: high staining) and a primary-antibody omission control (standard IHC practice).
Could fixation explain weak or uneven BMX staining?
BMX-specific sensitivity to fixation is unknown from the supplied evidence; the selected paraffin-section caption does not state its fixative (caption: A02537-1). Record the actual fixative and fixation duration for each specimen, and compare sections processed together before assigning a weak result to biology (standard IHC practice). Excessive or uneven fixation can alter antigen accessibility in IHC generally, so repeat staining with the specified citrate retrieval before changing several steps at once (standard IHC practice; page retrieval rule). Include a positive tissue control and a primary-antibody omission control in the same run to separate assay failure from nonspecific detection (standard IHC practice).
Is membrane staining compatible with a cytoplasmic BMX annotation?
Yes: UniProt places BMX in the cytoplasm and reports localization at spreading-cell edges when it complexes with BCAR1 (UniProt P51813). BMX has no transmembrane segment, so a membrane-associated pattern should not be described as proof that it spans the membrane (UniProt P51813 topology). HPA reports plasma membrane as its main subcellular location and nucleoplasm as an additional location in its subcellular assessment (HPA: subcellular). The selected paraffin-section image describes cytoplasmic and cell membrane staining at 1:50 (caption: A02537-1). Score these compartments separately, checking whether apparent edge staining follows cell boundaries or section folds (standard IHC practice).
How can I assess whether the antibody recognizes the intended BMX epitope?
The supplied record lists 0 isoforms, so it offers no isoform-specific staining pattern to interpret (UniProt P51813). BMX contains a PH domain at residues 4–111, an SH2 domain at 296–392 and a kinase domain at 417–675 (UniProt P51813). Its reported phosphotyrosines include residues 216, 224 and 566, but their presence does not establish what this antibody recognizes (UniProt P51813). Check the catalog antibody’s stated immunogen or epitope before interpreting staining as domain-specific or phosphorylation-specific, and use an independent antibody or target depletion for confirmation when available (standard IHC practice).
How should I adapt a BMX question for multiplex IF?
For IF/ICC, pair BMX with a marker identifying the cell population being assessed; endothelial cells are a biologically relevant population for BMX (UniProt P51813 tissue specificity). Choose fluorophores after viewing an unstained specimen in each channel, placing the signal of interest where tissue autofluorescence is least disruptive (standard IF practice). Because BMX has no transmembrane segment, test permeabilisation when seeking intracellular epitopes, and assess membrane-associated signal without assuming an extracellular epitope (UniProt P51813 topology; standard IF practice). The HPA subcellular assessment includes IF images from HUVEC/TERT2, but that observation does not establish an IF protocol for the catalog antibody (HPA: subcellular).
How do I distinguish BMX signal from chromogenic background?
Run a primary-antibody omission control beside each chromogenic IHC batch; the selected paraffin-section image used PBS in place of primary antibody for its negative control (caption: A02537-1). Check endogenous peroxidase blocking and inspect a section before DAB development if diffuse brown staining appears, treating these as general workflow checks (standard IHC practice). Compare suspected signal with tissue morphology and with a positive control containing epididymal glandular cells (HPA: high staining; standard IHC practice). If background rises with stronger retrieval, return to the page’s citrate pH 6.0, 20-minute condition and adjust one variable at a time (page retrieval rule; standard IHC practice).
What should I measure when comparing BMX staining across sections? ⚠ ANSWER MARKED FOR VERIFICATION
Define the cell population and compartment before scoring: HPA reports high staining in epididymal glandular cells, while UniProt and HPA describe cytoplasmic and membrane-associated localization (HPA: tissue and subcellular; UniProt P51813). For a defined population, report percentage of positive cells and an H-score combining staining intensity with that percentage; use positive-cell density per mm² when cell distribution is the question (standard IHC practice). Normalize counts to evaluable cells or viable tissue area, as appropriate, and exclude folds and necrotic regions consistently (standard IHC practice). Apply the same thresholds, imaging settings and control criteria to every section (standard IHC practice).
Which BMX staining patterns warrant a specificity check?
Interpret cytoplasmic or cell-edge staining in light of BMX’s cytoplasmic annotation and its reported association with spreading-cell edges (UniProt P51813). Membrane-associated signal is plausible, but the protein has no transmembrane segment; dominant staining in an unexpected compartment needs independent confirmation (HPA: subcellular; UniProt P51813 topology). Check the identity of stained cells against tissue morphology: HPA reports high staining in epididymal glandular cells and notes presumed off-target binding in its tissue assessment (HPA: tissue). Signal confined to cut edges, folds or necrosis, or reproduced in a primary-antibody omission control, should prompt a check for staining artefact or endogenous enzyme activity (standard IHC practice).
Boster reagents
Best BMX / Cytoplasmic tyrosine-protein kinase BMX IHC Antibodies
Two anti-BMX antibodies have paraffin-section IHC images in human carcinoma tissue (catalog image captions); both list IF and Human, Mouse, and Rat reactivity (catalog applications and reactivity).
Real IHC data
Anti-ETK (K35) BMX Antibody
Cat # A02537-1
Real IHC data
Anti-ETK (Y43) BMX Antibody
Cat # A02537Y43
A02537-1 has IHC images from paraffin-embedded human liver and rectum carcinoma tissue and lists IF (A02537-1 image captions; catalog applications). A02537Y43 has an IHC image from paraffin-embedded human tonsil carcinoma tissue and lists IF (A02537Y43 image caption; catalog applications).
Which to pick: For tissue IHC, choose A02537-1 when its liver or rectum carcinoma images match the planned comparison, or A02537Y43 for its tonsil carcinoma image; each caption documents paraffin sections, while the fixative is unreported (respective IHC image captions). For IF, either is listed at 1:50–1:200, but neither has an IF image in the payload, and ICC validation is unreported (catalog applications, dilutions, and image records). For cross-species work, both list Human, Mouse, and Rat reactivity, though their IHC images show human tissue only (catalog reactivity; respective IHC image captions).
Each figure is that product's own IHC / IF validation image from its datasheet.
References
- UniProt Consortium. UniProt entry P51813 (BMX_HUMAN, Cytoplasmic tyrosine-protein kinase BMX).
- Human Protein Atlas. BMX tissue IHC expression (reliability: Enhanced).
- Human Protein Atlas. BMX subcellular location (ICC-IF): Mainly localized to the plasma membrane. In addition localized to the nucleoplasm..
- Human Protein Atlas. BMX antibody validation summary (2 antibodies).
- BMX/Etk promotes cell proliferation and tumorigenicity of cervical cancer cells through PI3K/AKT/mTOR and STAT3 pathways. Oncotarget 2017 — PMC5564764.
- Single-Cell RNA Sequencing Reveals Atlas of Yak Testis Cells. International journal of molecular sciences 2023 — PMC10178277.
- BMX-Mediated Regulation of Multiple Tyrosine Kinases Contributes to Castration Resistance in Prostate Cancer. Cancer research 2018 — PMC6139052.
- BMX Represses Thrombin-PAR1-Mediated Endothelial Permeability and Vascular Leakage During Early Sepsis. Circulation research 2020 — PMC7035171.
- PubMed PMID:7970727 — UniProt-cited evidence.
- PubMed PMID:9520419 — UniProt-cited evidence.
- PubMed PMID:15772651 — UniProt-cited evidence.
Immunohistochemistry protocol sheet for BMX (Cytoplasmic tyrosine-protein kinase BMX). Expected localisation Cytoplasm in epididymal glandular cells (HPA tissue IHC). Antigen retrieval: Citrate pH 6.0 HIER, 95–98 °C, 20 min (rule: cytoplasmic / membrane antigen). Validated IHC antibody: A02537-1 at 1:50-1:200 (the product's own IHC validation image is shown). Controls (HPA): positive Epididymis (Glandular cells); negative Adipose tissue. IHC-P workflow on paraffin sections; fixative unspecified. Protocol steps: 1. Sample: Tissue sections; selected-image fixative not specified (standard IHC workflow); 2. Fixation: Image fixative and duration unreported (datasheet A02537-1); 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-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-BMX, 1:50-1:200 (datasheet A02537-1); 9. Primary incubation: Overnight at 4 °C (standard). Watch for: Fixation: Keep fixation consistent across paraffin sections (standard IHC practice; not target-specific); Caveat: Presumed off-target staining; verification pending (HPA tissue IHC); Isoform / epitope: No reported isoforms or processing; one cytoplasmic chain (UniProt); Regulation: Highly expressed in migratory cells (UniProt). Evidence: UniProt P51813, HPA tissue IHC · Enhanced, HPA ICC-IF · Mainly localized to the plasma membrane. In addition localized to…, Datasheet A02537-1, PMC5564764, PMC10178277. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/BMX/immunohistochemistry-ihc-guide.