This website uses cookies to ensure you get the best experience on our website.
- Table of Contents
BTRC / F-box/WD repeat-containing protein 1A · IHC design guide
Design Immunohistochemistry for BTRC
Plan BTRC chromogenic IHC in paraffin sections around the general cytoplasmic tissue pattern (HPA tissue IHC), while recognising that nuclear localisation is also annotated at the molecular level (UniProt). The IHC-validated antibody has a stated dilution range of 1:50–1:200 (datasheet: A01747).
Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
BTRC 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 | Predominantly cytoplasmic in tissue (HPA tissue IHC) | |
| Staining pattern | Cytoplasmic staining across multiple cell types (HPA tissue IHC) | |
| Antigen retrieval | Citrate pH 6.0 HIER, heat-mediated (datasheet A01747) | |
| Positive control | Appendix+4 more · see all | |
| Negative control | Spleen |
Important caveats
Reasons your staining may differ from the expected pattern.
| Fixation | Keep formalin fixation consistent across paraffin sections. (standard IHC practice; not target-specific) | |
| Caveat | Staining has medium consistency with RNA data (HPA tissue IHC) | |
| Regulation | Low tissue specificity; no defined inducer (HPA tissue IHC) | |
| Isoform / epitope | 2 isoforms; antibody epitope coverage is unspecified (UniProt) |
Section 1
Recommended BTRC IHC & IF Protocols
Start with the catalog antibody’s citrate pH 6.0 retrieval protocol (datasheet A01747). The published option below describes staining of FFPE sections (PMC13280727).
| Sample | Paraffin-embedded mouse spleen tissue; fixative not specified (datasheet A01747) |
| Fixation | Image fixative and duration unreported (datasheet A01747); verify before use. |
| Sectioning | 4–5 µm sections on charged slides (standard) |
| Deparaffinisation | Xylene, graded ethanol series to water (standard) |
| Antigen retrieval | Heat retrieval: Citrate pH 6.0 (datasheet A01747); 20 min, 95–100 °C (standard) |
| 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-BTRC, 1:50-1:200 (datasheet A01747) |
| 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 | BTRC-positive staining in glandular cells of appendix (HPA tissue IHC: Medium). HPA tissue profile: General cytoplasmic expression. No signal in the no-primary control. |
💡Decision noteTry heat-mediated citrate pH 6.0 retrieval first (datasheet A01747); the published FFPE protocol also uses citrate pH 6.0 after formic acid treatment (PMC13280727).
Section 2
What Is the Expected BTRC Staining Pattern?
BTRC should appear chiefly as cytoplasmic staining across several cell types in tissue IHC, including glandular, respiratory epithelial, glial, and endothelial cells (HPA tissue IHC: general cytoplasmic expression; medium staining in listed cells). Nuclear staining is biologically plausible (UniProt Q9Y297: cytoplasm and nucleus), but should be assessed against the tissue pattern. BTRC has no transmembrane segment (UniProt Q9Y297: topology). HPA rates the tissue IHC evidence Approved, with medium consistency between staining and RNA data (HPA tissue IHC: reliability).
| Moderate cytoplasmic signal in appendix or breast glandular cells, or bronchial respiratory epithelial cells. | This matches the reported tissue IHC pattern (HPA tissue IHC: Medium in these cells; general cytoplasmic expression). Judge signal in the named cells and compartment; staining elsewhere on the section does not by itself establish a BTRC-positive result. |
| Dominant nuclear-only or sharply membranous staining in a tissue section. | Check for an assay-dependent pattern or artefact before calling it positive: tissue IHC is described as generally cytoplasmic (HPA tissue IHC: profile). Nuclear localization remains possible (UniProt Q9Y297: nucleus); plasma-membrane localization was observed in ICC-IF, a different assay (HPA subcellular: additional location). |
| Strong signal in an unexpected cell population, especially when the expected cells are unstained. | Consider antibody cross-reactivity or endogenous chromogenic detection activity (standard IHC practice). HPA reports low tissue specificity and several stained cell types, so an unlisted population is a reason to verify specificity, not proof of a false positive (HPA tissue IHC: RNA specificity and positive-cell list). |
| Diffuse color across cells, stroma, or the whole section without a clear cellular pattern. | Treat this as background until controls resolve it (standard IHC practice). Excess detection signal, inadequate blocking, or insufficient washing can obscure compartment boundaries (standard IHC practice). A diffuse field cannot be matched confidently to HPA's general cytoplasmic profile (HPA tissue IHC: profile). |
| No signal in appendix glandular cells or bronchial respiratory epithelial cells. | First check section quality, retrieval, antibody incubation, and detection controls (standard IHC practice). These are useful reference cells because HPA reports Medium staining, although its Approved rating has only medium consistency with RNA data (HPA tissue IHC: positive cells and reliability). A failed reference section cannot establish that the test tissue lacks BTRC. |
💡Expected BTRC appearanceCall a result consistent with BTRC when identifiable reference cells show medium, mainly cytoplasmic chromogenic staining (HPA tissue IHC: positive-cell levels and profile); widespread structureless color or staining confined to an unexpected population warrants control-based review (standard IHC practice).
| Compartment and assay | Tissue IHC is generally cytoplasmic (HPA tissue IHC: profile); ICC-IF is mainly nucleoplasmic, with cytosol and plasma membrane also reported (HPA subcellular: approved locations). Interpret each assay against its own evidence. |
| Reference-cell selection | Appendix and breast glandular cells, bronchial epithelium, and selected neural or endothelial cells show Medium staining (HPA tissue IHC: positive cells). Spleen red-pulp cells are reported Not detected; that finding does not classify the entire spleen (HPA tissue IHC: negative cell). |
| Antibody validation | CAB032986 is Approved for IHC; HPA031156 is Approved for ICC, with no IHC status listed for it (HPA antibodies: assay-specific status). An Approved label is not an Enhanced validation claim (HPA antibodies: validation status). |
| Protein features | BTRC has two isoforms, no listed signal peptide or propeptide, and no transmembrane segment (UniProt Q9Y297: isoforms, processing, topology). The supplied record gives no antibody epitope, so isoform coverage cannot be inferred. |
| Evidence limit for paraffin sections | No target-specific fixation or retrieval effect is provided (supplied UniProt and HPA records). Retrieval may be optimized as general IHC practice, but these sources cannot predict which condition improves BTRC staining. |
| Situation | Likely cause | Next action |
|---|---|---|
| Reference cells are blank and the test section is blank. | A failed staining run or unsuitable assay conditions are possible (standard IHC practice); HPA reports Medium staining in several reference cells (HPA tissue IHC: positive cells). | Confirm the IHC antibody identity and assay validation, then check retrieval, incubation, detection reagents, and the positive control (standard IHC practice; HPA antibodies: CAB032986 IHC Approved). |
| Color is widespread, with little contrast between cells and surrounding tissue. | Nonspecific background or endogenous detection activity may overwhelm cellular signal (standard IHC practice). | Review the no-primary control; optimize blocking, washes, antibody concentration, and detection exposure as needed (standard IHC practice). Reassess whether identifiable cells show the HPA cytoplasmic pattern (HPA tissue IHC: profile). |
| Only nuclei stain in tissue IHC. | BTRC can occur in the nucleus (UniProt Q9Y297: subcellular location), but a nuclear-only result differs from HPA's general tissue IHC profile (HPA tissue IHC: profile). | Compare a reference tissue and appropriate controls before scoring; record the nuclear pattern separately from the expected cytoplasmic tissue IHC signal (standard IHC practice; HPA tissue IHC: profile). |
| Spleen red-pulp cells stain strongly. | This conflicts with the reported Not detected result for those cells (HPA tissue IHC: spleen red pulp); cross-reactivity or background is possible (standard IHC practice). | Verify cell identity and inspect no-primary and positive-reference controls (standard IHC practice). Do not extend the red-pulp finding to every spleen cell (HPA tissue IHC: negative cell). |
| Epididymis glandular cells stain weakly despite the UniProt tissue note. | UniProt reports protein expression in epididymis, while HPA lists Low glandular staining there (UniProt Q9Y297: tissue specificity; HPA tissue IHC: low group). | Use an HPA Medium-staining cell population as the run reference, and interpret weak epididymal staining within its reported range (HPA tissue IHC: positive and low groups). |
| ICC-IF shows nucleoplasmic signal while paraffin IHC looks cytoplasmic. | The reported profiles differ by assay (HPA subcellular: mainly nucleoplasmic ICC-IF; HPA tissue IHC: general cytoplasmic expression). The listed Approved ICC and IHC antibodies also differ (HPA antibodies: assay-specific status). | Evaluate ICC-IF against its separate guide and assay controls (standard ICC-IF practice); retain the tissue IHC profile when scoring paraffin sections (HPA tissue IHC: profile). |
Experiment evidence
IHC & IF images showing BTRC staining
A01747 · IHCImmunohistochemistry of paraffin-embedded mouse spleen using β-TrCP/BTRC Rabbit pAb at dilution of 1:100 .Perform high pressure antigen retr
A01747 · IFImmunofluorescence analysis of C6 cells using β-TrCP/BTRC Rabbit pAb at dilution of 1:100 . Blue: DAPI for nuclear staining.
HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000166167-BTRC/tissue/cerebral+cortex#imid_8170400
HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000166167-BTRC/tissue/colon#imid_8170318
HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000166167-BTRC/tissue/liver#imid_8170319
HPA · KidneyHuman Protein Atlas IHC, Kidney. Source: https://www.proteinatlas.org/ENSG00000166167-BTRC/tissue/kidney#imid_8170336
HPA · ICC-IF A-431Human Protein Atlas immunofluorescence, A-431, antibody HPA031156; green = BTRC, blue = nucleus, red = microtubules.
HPA · ICC-IF A-431Human Protein Atlas immunofluorescence, A-431, antibody HPA031156; green = BTRC, blue = nucleus, red = microtubules.
← swipe / scroll for more images →
Real IHC / IF data behind the staining-pattern expectations in this section.
Sample controls for BTRC IHC & IF
🧪Run appendix first: glandular cells should stain (HPA: Medium in appendix glandular cells). Use spleen red pulp as the HPA negative comparator (HPA: Not detected in spleen red-pulp cells); on the appendix slide, use non-glandular cells that remain unstained as internal background references, without assigning them a known BTRC-negative identity.
Positive control tissue: Appendix (Glandular cells, HPA Medium)
Negative control tissue: Spleen (HPA Not detected)
ICC-IF cell lines (HPA subcellular resource): HPA ICC-IF images show BTRC in A-431, U-251MG, U2OS, with annotated localisation: Nucleoplasm (approved) (HPA subcellular).
Technical controls: Include a no-primary (secondary-only) control, a concentration-matched rabbit polyclonal isotype control (selected-SKU caption: rabbit pAb), and BTRC-knockout tissue as a biological negative where available (standard IHC controls). Quench endogenous peroxidase and check inflammatory cells in appendix for residual chromogen background (standard chromogenic IHC practice).
⚠️Feasibility: A target-specific fixation window or fixation effect is unreported in the supplied evidence; the selected-SKU mouse-spleen paraffin caption also does not report its fixative (selected-SKU caption). That caption reports high-pressure retrieval in 10 mM citrate, pH 6.0, and a 1:100 antibody dilution, but does not establish that retrieval is required for other tissues (selected-SKU caption). The documented paraffin IHC starting point is more concrete than a frozen-section or IF protocol; interpret spleen cautiously because the mouse-spleen caption shows staining while HPA reports no detection in spleen red-pulp cells (selected-SKU caption; HPA: Not detected in spleen red-pulp cells).
HPA tissue IHC evidence for BTRC
Comprehensive Human Protein Atlas IHC scoring per tissue (reliability: Approved — Medium consistency between antibody staining and RNA expression data.). 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 |
|---|---|---|---|---|
| Appendix | Glandular cells | Medium | Protein (IHC) | HPA → |
| Breast | Glandular cells | Medium | Protein (IHC) | HPA → |
| Bronchus | Respiratory epithelial cells | Medium | Protein (IHC) | HPA → |
| Caudate | Glial cells | Medium | Protein (IHC) | HPA → |
| Cerebellum | Cells in granular layer | Medium | Protein (IHC) | HPA → |
Undetected expression · recommended negative controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| Spleen | Cells in red pulp | Not detected | Protein (IHC) | HPA → |
Section 3
Advanced BTRC IHC Tips
Use compartment, cell type, and matched controls to interpret BTRC staining in paraffin sections; the selected tissue image reports retrieval conditions but not fixation.
What retrieval should I try first for weak BTRC staining in paraffin sections?
Start with heat-mediated retrieval in 10 mM citrate buffer, pH 6.0 (datasheet A01747; selected IHC caption). The selected mouse spleen image used high-pressure retrieval and a 1:100 primary dilution, so use those as an initial reference while recording your heating and cooling conditions (selected IHC caption). If staining remains weak, adjust retrieval time in small steps and compare adjacent sections with identical detection conditions; excessive heating can damage morphology (standard IHC practice). Judge improvement by cell-associated staining and preserved tissue structure, rather than by a general increase in brown signal; include a primary-antibody omission control to expose detection background (standard IHC practice).
How should I troubleshoot fixation-related loss of BTRC staining?
BTRC-specific sensitivity to fixation is unknown from the supplied evidence; the selected caption identifies paraffin-embedded mouse spleen but does not state its fixative (selected IHC caption). Record the fixative, fixation duration, tissue thickness, and processing history for each specimen before comparing staining intensity (standard IHC practice). If archived blocks vary, stain adjacent sections together with the same 10 mM citrate, pH 6.0 retrieval and antibody conditions, then compare morphology and signal (datasheet A01747; standard IHC practice). Test a controlled fixation series only when comparable fresh material is available, and avoid attributing a weak result to fixation until retrieval, detection, and tissue preservation have been checked (standard IHC practice).
Should BTRC staining be cytoplasmic, nuclear, or membrane-associated?
Assess cytoplasmic and nuclear staining separately: UniProt places BTRC in both compartments, while HPA tissue IHC describes general cytoplasmic expression (UniProt Q9Y297 subcellular location; HPA tissue IHC). HPA cell imaging places it mainly in the nucleoplasm, with additional cytosol and plasma-membrane localisation, so those observations can guide review without dictating every tissue pattern (HPA subcellular). BTRC has no predicted transmembrane segment, making a sharp, exclusively membranous chromogenic rim a pattern to investigate carefully (UniProt Q9Y297 topology). Compare compartments in well-preserved cells, inspect the counterstain for nuclear boundaries, and exclude folds or section edges before recording a compartment-specific score (standard IHC practice).
Can this antibody distinguish BTRC isoforms or reveal an inaccessible epitope?
The record lists 2 BTRC isoforms, but the supplied antibody evidence does not map its epitope or establish isoform selectivity (UniProt Q9Y297 isoforms; selected IHC caption). Therefore, report staining as BTRC immunoreactivity unless independent epitope information and appropriate controls support an isoform-specific claim (standard IHC interpretation). BTRC contains an F-box domain at residues 190–228 and WD repeats, yet those features alone cannot locate this antibody's binding site (UniProt Q9Y297 domains and keywords). If retrieval changes the pattern, compare adjacent sections and control tissues under matched conditions; a stronger signal alone cannot identify which isoform or epitope became accessible (standard IHC practice).
How can I check the IHC pattern with multiplex IF?
For a follow-up IF experiment, pair BTRC with a marker for the cell population being evaluated in the IHC section, and inspect each channel independently before interpreting overlap (standard IF practice). Choose fluorophores after checking unstained tissue for autofluorescence; place a weaker target signal in a cleaner spectral channel and include single-stain controls (standard IF practice). Because BTRC is reported in the cytoplasm and nucleus, assess whether permeabilisation permits access to those intracellular compartments, while treating the antibody's unmapped epitope as unknown (UniProt Q9Y297 subcellular location; selected IHC caption). Compare the IF compartment pattern with the chromogenic result, without assuming that fluorescence intensity and brown deposit measure abundance identically (standard IHC/IF practice).
How do I separate BTRC staining from chromogenic background?
Run a section without primary antibody to reveal secondary-reagent or detection background, and inspect areas with endogenous pigment before assigning brown deposits to BTRC (standard IHC practice). Include a peroxidase block when using an enzyme-based DAB system; this is a general chromogenic workflow step, not evidence about BTRC expression (standard IHC practice). If diffuse staining obscures cells, optimise blocking, washes, primary concentration, and DAB development one variable at a time; 1:100 is the selected image's antibody dilution (selected IHC caption; standard IHC practice). Prefer reproducible cell-associated signal over edge staining, precipitate, or deposits that also appear in the primary-omission control (standard IHC practice).
What is a defensible way to quantify BTRC IHC across specimens? ⚠ ANSWER MARKED FOR VERIFICATION
Define the cell population and compartment before scoring, because BTRC can occupy cytoplasmic and nuclear compartments and HPA tissue IHC emphasizes cytoplasmic staining (UniProt Q9Y297 subcellular location; HPA tissue IHC). For intact cells, report the percentage positive and an H-score calculated as 1× weak + 2× moderate + 3× strong percentage staining, keeping nuclear and cytoplasmic scores separate (standard IHC practice). For spatial counts, report positive cells per mm² of viable, annotated tissue and retain the denominator for each specimen (standard IHC practice). Apply one threshold across a staining batch, normalise to the same cell population or viable area, and exclude folds, necrosis, and section loss (standard IHC practice).
What makes a BTRC-positive IHC result convincing?
Look for reproducible staining in intact cells with interpretable cytoplasmic or nuclear localisation, consistent with the reported BTRC compartments (UniProt Q9Y297 subcellular location; HPA tissue IHC). Check cell identity against the section's morphology and relevant markers: HPA reports medium staining in several specified cell populations but does not make every cell in those tissues positive (HPA tissue IHC). Treat an isolated edge accent, necrotic area, or brown signal persisting without primary antibody as possible artefact, and review peroxidase blocking when using DAB (standard IHC practice). The selected mouse spleen image and HPA's undetected human spleen red-pulp entry concern different specimens, so neither alone establishes universal spleen positivity or negativity (selected IHC caption; HPA tissue IHC).
Boster reagents
Best BTRC / F-box/WD repeat-containing protein 1A IHC Antibodies
A01747 has IHC data from paraffin-embedded mouse spleen and IF data from C6 cells (catalog image captions); listed reactivity includes human, mouse, and rat (catalog reactivity).
Real IHC data
Anti-BTRC Antibody
Cat # A01747
A01747 will render with an IHC image of paraffin-embedded mouse spleen stained at 1:100 after citrate retrieval (A01747 IHC image caption). A01747 also has an IF image of C6 cells stained at 1:100 with DAPI nuclear staining (A01747 IF image caption).
Which to pick: Choose A01747 for paraffin-section IHC; its own mouse spleen image documents that preparation, while the fixative is unreported (A01747 IHC image caption). Choose A01747 for IF/ICC because both applications are listed and its IF image shows C6 cells (catalog applications; A01747 IF image caption). For work across species, A01747 is listed as reactive with human, mouse, and rat and is described as a rabbit polyclonal antibody in its IHC caption (catalog reactivity; A01747 IHC image caption).
Each figure is that product's own IHC / IF validation image from its datasheet.
References
- UniProt Consortium. UniProt entry Q9Y297 (FBW1A_HUMAN, F-box/WD repeat-containing protein 1A).
- Human Protein Atlas. BTRC tissue IHC expression (reliability: Approved).
- Human Protein Atlas. BTRC subcellular location (ICC-IF): Mainly localized to the nucleoplasm. In addition localized to the plasma membrane and cytosol..
- Human Protein Atlas. BTRC antibody validation summary (2 antibodies).
- miR-224 targets BTRC and promotes cell migration and invasion in colorectal cancer. 3 Biotech 2020 — PMC7585582.
- MGMT genomic rearrangements contribute to chemotherapy resistance in gliomas. Nature communications 2020 — PMC7403430.
- Double Chromogen-based Immunohistochemical Staining: An Efficient Approach for Utilizing Long-term Formalin-fixed Tissue in Biobanks. Applied immunohistochemistry & molecular morphology : AIMM 2024 — PMC13280727.
- EBV-miR-BART10-3p facilitates epithelial-mesenchymal transition and promotes metastasis of nasopharyngeal carcinoma by targeting BTRC. Oncotarget 2015 — PMC4747187.
- PubMed PMID:9859996 — UniProt-cited evidence.
- PubMed PMID:9660940 — UniProt-cited evidence.
- PubMed PMID:10531035 — UniProt-cited evidence.
Immunohistochemistry protocol sheet for BTRC (F-box/WD repeat-containing protein 1A). Expected localisation Predominantly cytoplasmic in tissue (HPA tissue IHC). Antigen retrieval: Citrate pH 6.0 HIER, heat-mediated (datasheet A01747). Validated IHC antibody: A01747 at 1:50-1:200 (the product's own IHC validation image is shown). Controls (HPA): positive Appendix (Glandular cells); negative Spleen. IHC workflow; specimen preparation unspecified. Protocol steps: 1. Sample: Paraffin-embedded mouse spleen tissue; fixative not specified (datasheet A01747); 2. Fixation: Image fixative and duration unreported (datasheet A01747); 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: Citrate pH 6.0 (datasheet A01747); 20 min, 95–100 °C (standard); 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-BTRC, 1:50-1:200 (datasheet A01747); 9. Primary incubation: Overnight at 4 °C (standard). Watch for: Fixation: Keep formalin fixation consistent across paraffin sections. (standard IHC practice; not target-specific); Caveat: Staining has medium consistency with RNA data (HPA tissue IHC); Isoform / epitope: 2 isoforms; antibody epitope coverage is unspecified (UniProt); Regulation: Low tissue specificity; no defined inducer (HPA tissue IHC). Evidence: UniProt Q9Y297, HPA tissue IHC · Approved, HPA ICC-IF · Mainly localized to the nucleoplasm. In addition localized to the…, Datasheet A01747, PMC13280727. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/BTRC/immunohistochemistry-ihc-guide.