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BMX / Cytoplasmic tyrosine-protein kinase BMX · IHC design guide
Design Immunohistochemistry for BMX
Plan chromogenic BMX IHC around the cytoplasmic pattern in epididymal glandular cells (HPA tissue IHC) and an IHC-validated antibody diluted 1:50–1:200 (datasheet: A02537-1). Include a no-primary control and interpret staining with the reported presumed off-target binding in mind (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 | Cytoplasmic staining in epididymal glandular cells (HPA tissue IHC) | |
| Staining pattern | Distinct cytoplasmic signal 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 sections (standard IHC practice; not target-specific). Selected-image fixative and duration unreported (datasheet A02537-1); verify before use. | |
| Caveat | Presumed off-target binding may confound staining (HPA tissue IHC) | |
| Regulation | High in cells with migratory potential (UniProt) | |
| Isoform / epitope | No isoforms; one 1–675 chain; epitope site unknown (UniProt) |
Section 1
Recommended BMX IHC & IF Protocols
The catalog antibody’s IHC-P protocol is paired with two published BMX IHC protocols (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 retrieval specification). A published pressure-cooker protocol uses citrate for 2 min (PMC5564764).
Section 2
What Is the Expected BMX Staining Pattern?
In paraffin-section IHC, the clearest expected BMX signal is distinct cytoplasmic staining in epididymal glandular cells, with medium staining reported in testicular Sertoli cells (HPA: tissue IHC). BMX is a cytoplasmic kinase without a transmembrane segment (UniProt P51813: location and topology). HPA rates its tissue IHC evidence Enhanced, while reporting medium staining–RNA consistency, presumed off-target binding that was disregarded, and pending external verification (HPA: reliability).
| Distinct cytoplasmic staining in epididymal glandular cells, with comparatively weaker staining in Sertoli cells. | This follows the reported high epididymal and medium Sertoli-cell patterns (HPA: tissue IHC). Read intensity relative to controls on the same run; absolute darkness alone does not establish specificity (general IHC practice). |
| Predominantly membranous or nuclear staining replaces the expected cytoplasmic pattern in tissue sections. | Review before calling it BMX: tissue IHC reports cytoplasm (HPA: tissue IHC). Membrane localization is supported and nucleoplasmic localization is approved in ICC-IF (HPA: subcellular), so those findings alone do not prove artefact; the assay context matters. |
| Strong staining appears in cells recorded as not detected, such as adipocytes or bone-marrow hematopoietic cells. | This conflicts with the sampled tissue IHC pattern (HPA: tissue IHC). Consider nonspecific antibody staining or endogenous detection activity (general IHC practice); compare with a detection-only control before assigning a cause. |
| Broad, diffuse color covers tissue and spaces between cells rather than outlining individual cytoplasms. | That distribution cannot be read confidently as the distinct epididymal cytoplasmic signal (HPA: tissue IHC). Excess detection background, inadequate blocking, or incomplete washing are possible general IHC causes (general IHC practice). |
| The epididymal glandular-cell positive control has no convincing cytoplasmic staining. | A negative study slide is uninterpretable if the reported high-staining control also fails (HPA: High in epididymal glandular cells; general IHC practice). Check control tissue, antibody and detection performance before concluding that the study sample lacks BMX. |
💡Expected BMX appearanceCall a run positive when glandular cells show high, distinct cytoplasmic staining in epididymis (HPA: tissue IHC); widespread diffuse color or strong staining in recorded negative cells warrants specificity checks (HPA: tissue IHC; general IHC practice).
| Tissue and cell choice | Epididymal glandular cells provide the strongest reported tissue IHC reference; Sertoli cells are medium, while several listed cell populations are not detected (HPA: tissue IHC). Score the named cells, not the whole section. |
| Assay-dependent localization | Tissue IHC describes distinct cytoplasm, whereas ICC-IF supports plasma membrane localization and also reports nucleoplasm (HPA: tissue IHC and subcellular). Use the tissue IHC pattern for this paraffin-section guide. |
| Validation limits | Two listed antibodies have Enhanced IHC status (HPA: antibody validation). HPA also notes presumed off-target binding and pending external verification, so agreement with the reference pattern still needs controls (HPA: reliability). |
| Protein organization | BMX has no transmembrane segment and is annotated as cytoplasmic; localization at spreading-cell edges is reported when complexed with BCAR1 (UniProt P51813: topology and location). This does not specify a paraffin-section edge-staining pattern. |
| Protocol and epitope uncertainty | The supplied UniProt and HPA records give no antibody epitope or BMX-specific antigen-retrieval result (UniProt P51813; HPA: supplied records). Treat retrieval conditions as an assay optimization choice, without predicting a BMX-specific fixation effect. |
| Situation | Likely cause | Next action |
|---|---|---|
| No signal in both the study section and epididymal positive control. | The reported high-staining glandular-cell reference has failed (HPA: High in epididymis). Antibody, detection, tissue preservation, or retrieval workflow problems remain possible (general IHC practice). | Confirm that glandular cells are present; check reagent application and detection controls, then follow the antibody's validated IHC-P conditions where available (general IHC practice). Do not score the study section as BMX-negative yet. |
| Epididymis stains, but the study tissue is negative. | Expression may differ by sampled cell population; HPA reports several populations as not detected (HPA: tissue IHC). A working positive control does not establish expression in every tissue (general IHC practice). | Identify the cell population being scored and report its result against the stained control. Avoid extrapolating UniProt's endothelial or migratory-cell expression statement to a particular IHC section (UniProt P51813: tissue specificity). |
| Strong signal appears in an HPA negative reference population. | That disagrees with the reported not-detected pattern (HPA: tissue IHC); nonspecific binding or endogenous chromogen-generating activity is possible (general IHC practice). | Compare a detection-only section, review blocking of endogenous activity, and assess whether the color is cell-associated (general IHC practice). HPA's Enhanced rating does not remove its off-target caveat (HPA: reliability). |
| Color is diffuse across the section or obscures cell boundaries. | The slide cannot resolve the distinct cytoplasmic pattern reported for epididymal glandular cells (HPA: tissue IHC). Concentrated reagent, incomplete washing, or nonspecific detection can contribute (general IHC practice). | Compare with the negative detection control; review blocking, washes, antibody concentration, and detection development using validated IHC-P guidance (general IHC practice). Reassess compartment only after background is controlled. |
| Membrane or nuclear signal dominates the paraffin-section result. | It differs from the tissue IHC cytoplasmic profile (HPA: tissue IHC). ICC-IF separately reports supported plasma membrane and approved nucleoplasmic localization, so the compartment alone cannot identify the cause (HPA: subcellular). | Compare the same run's epididymal control and detection-only control, then describe compartment and cell type explicitly (general IHC practice). Do not use the ICC-IF localization record as an IHC-P protocol or as proof of tissue-section specificity. |
| Only faint, scattered cells stain in the epididymal control. | The result falls short of the reported high glandular-cell staining, although section composition and assay performance can affect interpretation (HPA: tissue IHC; general IHC practice). No BMX-specific fixation sensitivity is supplied (UniProt P51813; HPA: supplied records). | Check that glandular epithelium is represented, inspect control and detection performance, and compare retrieval and dilution with the antibody's validated IHC-P instructions if available (general IHC practice). Do not infer a BMX-specific fixation cause. |
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 and score its glandular cells, where BMX staining is High (HPA: epididymis glandular cells, High). Use adrenal gland glandular cells as a negative tissue (HPA: adrenal gland glandular cells, Not detected); on the epididymis slide, treat cells outside the scored glandular compartment as internal background references only if they are unstained, rather than assuming they are BMX-negative (HPA: epididymis glandular cells, High).
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 a no-primary, secondary-only control, concentration-matched nonimmune rabbit IgG for the rabbit polyclonal antibody, and BMX-knockout tissue or cells as a biological negative where available (hero caption: rabbit pAb and goat anti-rabbit secondary; standard IHC practice). Block endogenous peroxidase and check for endogenous biotin when using the caption’s avidin–peroxidase detection system (hero caption: avidin–peroxidase; standard IHC practice).
⚠️Feasibility: No target-specific fixation window, fixation effect, or antigen-retrieval dependency is reported in the supplied evidence; the selected A02537-1 paraffin-section caption does not state a fixative (hero caption: paraffin section; fixative unreported). The evidence does not establish whether frozen sections or IF/ICC would be easier, although HUVEC/TERT2 ICC-IF images are available (HPA: HUVEC/TERT2 ICC-IF images). In epididymis, inspect glandular luminal material for nonspecific chromogen deposition when scoring cells (HPA: epididymis glandular cells, High; standard IHC practice).
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
Use the paraffin-section image and tissue-expression data as controls while evaluating BMX staining by chromogenic IHC.
How should I retrieve BMX in paraffin sections when cytoplasmic staining is weak?
Start with citrate buffer at pH 6.0, using heat-induced epitope retrieval at 95–98 °C for 20 min (page retrieval rule). The catalog antibody has a paraffin-section liver carcinoma image at 1:50, but its caption does not report a retrieval method or fixative (A02537-1 caption). If staining remains weak, compare a second retrieval condition on adjacent sections while holding antibody concentration and detection constant (standard IHC practice). Check epididymal glandular cells as a positive tissue reference and adipocytes as a negative reference (HPA tissue IHC). Score cytoplasmic and membrane signal separately, since both appear in the catalog image (A02537-1 caption).
Can fixation explain a missing BMX signal in my IHC sections?
Target-specific fixation sensitivity for BMX is unknown from the supplied evidence (A02537-1 caption). Although the catalog image uses paraffin-embedded liver carcinoma tissue, its caption does not state the fixative or fixation duration (A02537-1 caption). Document those variables for each specimen and compare similarly processed sections before attributing a weak result to BMX abundance (standard IHC practice). Review tissue preservation and staining in a positive reference alongside the test section; epididymal glandular cells show high staining in the tissue dataset (HPA tissue IHC). If fixation differs across samples, optimize retrieval on matched material and report the processing difference when interpreting staining intensity (standard IHC practice).
Should BMX staining look cytoplasmic, membranous, or nuclear?
Expect cytoplasmic signal as one plausible IHC pattern because BMX is annotated in the cytoplasm (UniProt P51813 subcellular location). The catalog paraffin-section image reports cytoplasmic and cell membrane staining, while the subcellular dataset supports plasma-membrane localization and additionally reports nucleoplasm localization (A02537-1 caption; HPA subcellular). BMX has no transmembrane segment, so a membrane rim alone does not establish that it spans the membrane (UniProt P51813 topology). Record cytoplasmic, membrane-associated, and nuclear staining separately rather than combining them into one positive score (standard IHC practice). Evaluate each pattern against omission controls and tissue morphology, especially where section edges produce apparent rims (standard IHC practice).
How do BMX domains and phosphorylation affect antibody interpretation in IHC?
The supplied record lists no BMX isoforms and maps the PH domain to residues 4–111, SH2 domain to 296–392, and kinase domain to 417–675 (UniProt P51813 record). It also lists phosphorylation at Tyr 216, 224, and 566 (UniProt P51813 modified residues). The catalog caption does not identify the antibody epitope, so neither domain specificity nor phosphorylation dependence can be assigned to its staining (A02537-1 caption). If samples disagree, first compare retrieval, antibody concentration, and control staining on adjacent sections (standard IHC practice). Treat any proposed epitope or modification explanation as unresolved until an epitope-mapped reagent or suitable validation experiment supports it (standard IHC practice).
How can IF help assess BMX localisation seen by chromogenic IHC?
Use IF as a separate localization check when chromogenic IHC cannot distinguish closely apposed cells or compartments (standard microscopy practice). Multiplex BMX with a validated endothelial-cell marker where that cell population is relevant, because BMX is reported in endothelial cells with migratory potential (UniProt P51813 tissue specificity). Choose a red or far-red fluorophore after examining tissue autofluorescence in the intended channel (standard IF practice). Because BMX lacks a transmembrane segment and is annotated in the cytoplasm, test mild permeabilisation for access to intracellular epitopes; the antibody’s precise epitope is unreported (UniProt P51813 topology and subcellular location; A02537-1 caption). Compare compartment patterns with the IHC result, while keeping the IF preparation and its controls distinct (standard IF practice).
What controls help distinguish BMX staining from chromogenic background?
Include a no-primary section: the catalog image used PBS instead of primary antibody as its negative control (A02537-1 caption). Its detection used a biotinylated secondary antibody followed by avidin-peroxidase, so evaluate nonspecific secondary or detection-system staining on the control section (A02537-1 caption; standard IHC practice). With peroxidase and DAB detection, block endogenous peroxidase and inspect pigment or precipitate before calling a deposit BMX-positive (standard chromogenic IHC practice). Reduce antibody concentration or detection time only after confirming that a positive reference retains interpretable signal (standard IHC practice). Epididymal glandular cells provide a high-staining reference, whereas adipocytes are reported as not detected (HPA tissue IHC).
How should I score BMX staining across different cell populations? ⚠ ANSWER MARKED FOR VERIFICATION
Define the cell population and compartment before scoring, since the catalog image shows cytoplasmic and membrane staining and the tissue dataset identifies specific stained cell types (A02537-1 caption; HPA tissue IHC). For a specified population, report percent positive cells and an H-score from 0–300 using prespecified intensity categories (standard IHC scoring practice). Alternatively, report positive-cell density per mm² of viable tissue when cell distribution is the endpoint (standard image-analysis practice). Normalize counts to the number of eligible cells or measured viable area, and apply one threshold across comparably processed sections (standard IHC practice). Keep membrane and cytoplasmic scores separate and document excluded necrotic or damaged regions (standard IHC practice).
When is an apparent BMX-positive region more likely to be artefact?
Give greatest weight to staining in intact cells with plausible cytoplasmic or membrane-associated patterns, both reported for BMX in the catalog paraffin image (A02537-1 caption). Check the cell identity: high epididymal glandular-cell staining and absent adipocyte staining offer contrasting tissue references, although the tissue dataset notes presumed off-target binding (HPA tissue IHC). Treat isolated section-edge rims, necrotic deposits, and signal reproduced in a no-primary control as possible artefacts (standard IHC practice; A02537-1 caption). Nuclear signal warrants separate assessment because nucleoplasm localization is reported by subcellular imaging, but that finding does not validate nuclear chromogenic staining in a given section (HPA subcellular; standard IHC interpretation). For peroxidase detection, examine endogenous enzyme activity before assigning brown signal to BMX (standard chromogenic IHC practice).
Boster reagents
Best BMX / Cytoplasmic tyrosine-protein kinase BMX IHC Antibodies
Both anti-BMX antibodies have paraffin-section IHC images and are listed for IF, with Human, Mouse and Rat reactivity (IHC image captions; catalog applications/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 paraffin-section IHC images from human liver and rectum carcinoma, and is listed for IF (A02537-1 image captions; catalog applications). A02537Y43 has a paraffin-section IHC image from human tonsil carcinoma and is listed for IF (A02537Y43 image caption; catalog applications).
Which to pick: For tissue IHC, choose A02537-1 when its documented cytoplasmic and cell-membrane staining and PBS primary-omission control are useful; choose A02537Y43 when a tonsil carcinoma IHC example is more relevant (A02537-1 image captions; A02537Y43 image caption). For IF, either rabbit polyclonal antibody is catalog-listed; the payload provides no IF image or ICC validation (catalog host, dilution_raw, applications and image alts). Both list Human, Mouse and Rat reactivity, while their IHC captions specify paraffin sections without reporting a fixative (catalog reactivity; 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 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). 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 sections (standard IHC practice; not target-specific). Selected-image fixative and duration unreported (datasheet A02537-1); verify before use.; Caveat: Presumed off-target binding may confound staining (HPA tissue IHC); Isoform / epitope: No isoforms; one 1–675 chain; epitope site unknown (UniProt); Regulation: High in cells with migratory potential (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.