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BRI3BP / BRI3-binding protein · IHC design guide
Design Immunohistochemistry for BRI3BP
Plan BRI3BP IHC in paraffin sections using the granular cytoplasmic tissue pattern (HPA tissue IHC). The catalog antibody has an IHC dilution range of 1:100–1:300 (datasheet); glandular and glial cells show medium staining, while adipocytes are undetected (HPA tissue IHC).
Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
BRI3BP 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 | Granular cytoplasm (HPA tissue IHC); mitochondrial outer membrane expected (UniProt) | |
| Staining pattern | Granular cytoplasmic staining in most tissues (HPA tissue IHC) | |
| Antigen retrieval | Citrate pH 6.0 HIER, 95–98 °C, 20 min (rule: cytoplasmic / membrane antigen) | |
| Positive control | Adrenal gland+4 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 | Only medium consistency between IHC and RNA (HPA tissue IHC) | |
| Regulation | Overexpressed in various carcinomas (UniProt) | |
| Isoform / epitope | No annotated isoforms; epitope side across 4 membrane segments is unknown (UniProt) |
Section 1
Recommended BRI3BP IHC & IF Protocols
The catalog antibody’s IHC-P protocol is accompanied by published FFPE BRI3BP protocols for invasive breast cancer (PMC11730447) and hepatocellular carcinoma (PMC12578974).
| Sample | Paraffin-embedded human brain tissue; fixative not specified (datasheet A30536) |
| Fixation | Image fixative and duration unreported (datasheet A30536); 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-BRI3BP, 1:100 - 1:300 (datasheet A30536) |
| 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 | BRI3BP-positive staining in glandular cells of adrenal gland (HPA tissue IHC: Medium). HPA tissue profile: Granular cytoplasmic expression in most tissues. No signal in the no-primary control. |
💡Decision noteTry citrate pH 6.0 retrieval at 95–98 °C for 20 min first (page retrieval rule); both articles used citrate pH 6 (PMC11730447; PMC12578974).
Section 2
What Is the Expected BRI3BP Staining Pattern?
For paraffin-section IHC, expect granular cytoplasmic BRI3BP staining in many tissues, including glandular cells in colon and breast and glial cells in cerebral cortex (HPA: tissue IHC). BRI3BP is a four-pass mitochondrial outer-membrane protein (UniProt Q8WY22: topology and location). HPA rates the tissue staining Approved, with medium consistency between staining and RNA expression (HPA: tissue IHC reliability).
| Granular cytoplasmic signal in colon glandular cells. | This fits the reported IHC pattern and cell type; HPA records medium staining in colon glandular cells (HPA: tissue IHC). Judge intensity against a control processed in the same run (general IHC practice). |
| Predominantly nuclear, membranous, or uniformly diffuse staining on the IHC slide. | This differs from HPA's granular cytoplasmic tissue-IHC profile; check morphology, counterstain, and control sections before assigning localisation (HPA: tissue IHC; general IHC practice). Nuclear signal alone cannot be dismissed across assays: HPA also reports approved nucleoplasmic localisation by ICC-IF (HPA: subcellular). |
| Strong signal in adipocytes or squamous epithelial cells of cervix. | Those cells are reported as not detected by HPA tissue IHC; unexpected staining warrants a specificity and endogenous-detection check (HPA: tissue IHC; general IHC practice). It does not, by itself, establish cross-reactivity. |
| Chromogen spreads across cells, stroma, or the slide without a granular cellular pattern. | Treat this as background until controls show otherwise: compare a primary-antibody-omission control and inspect washing, blocking, and detection steps (general IHC practice). HPA describes granular cytoplasmic expression, not a diffuse field-wide pattern (HPA: tissue IHC). |
| No staining in a colon glandular-cell control. | HPA reports medium staining there, so a blank control raises a run-performance or sample-quality question; review the control's morphology and detection steps before calling the study sample negative (HPA: tissue IHC; general IHC practice). |
💡Expected BRI3BP appearanceCall a positive result when identifiable glandular cells show granular cytoplasmic chromogen of roughly medium intensity in a colon control; diffuse field-wide colour or strong staining in HPA-undetected adipocytes is suspect (HPA: tissue IHC).
| Subcellular structure | UniProt places BRI3BP in the mitochondrial outer membrane and annotates four transmembrane segments at residues 13–33, 125–145, 146–166, and 185–205 (UniProt Q8WY22: topology). HPA's granular cytoplasmic tissue pattern is consistent with an organelle-associated signal, but routine chromogenic IHC does not resolve individual mitochondria (HPA: tissue IHC; general IHC practice). |
| Tissue and cell selection | HPA reports medium staining in colon and breast glandular cells, cerebral-cortex glial cells, and bone-marrow hematopoietic cells; adipocytes and cervical squamous epithelial cells are not detected (HPA: tissue IHC). Select controls by the recorded cell population, rather than assuming every cell in a positive tissue must stain. |
| Strength of tissue evidence | The tissue-IHC profile is Approved, with medium consistency between antibody staining and RNA expression (HPA: tissue IHC reliability). HPA lists antibody HPA014957 as Approved for IHC, rather than Enhanced (HPA: antibody validation). Interpret unexpected compartments or cell types with controls rather than treating this profile as definitive. |
| Epitope and processing information | UniProt annotates a full-length 1–251 chain, no signal peptide or propeptide, and no glycosylation sites (UniProt Q8WY22: processing). The supplied record does not locate the antibody epitope, so the topology cannot establish which retrieval condition will expose it. |
| IF/ICC Q: Should its pattern match chromogenic tissue IHC? | A: HPA reports approved nucleoplasmic and mitochondrial localisation in ICC-IF images from RT-4, while its tissue-IHC summary is granular cytoplasmic (HPA: subcellular; HPA: tissue IHC). Compare each assay with its own reported pattern; this IHC section does not prescribe an IF protocol. |
| Situation | Likely cause | Next action |
|---|---|---|
| Colon glandular-cell control is blank. | The expected medium signal is absent; tissue preservation, retrieval, primary-antibody incubation, or detection may need review (HPA: tissue IHC; general IHC practice). | Confirm glandular cells are present, then inspect the run controls and each IHC step; repeat with a documented retrieval condition if the run failed (general IHC practice). |
| Signal is diffuse across cells and stroma. | Nonspecific binding, incomplete washing, or detection background can obscure a granular pattern (general IHC practice). | Check a primary-antibody-omission control; review blocking, wash steps, and chromogen development before scoring cells (general IHC practice). |
| Nuclear-only staining dominates the IHC section. | It conflicts with the granular cytoplasmic tissue-IHC summary, although nucleoplasm is an approved ICC-IF location (HPA: tissue IHC; HPA: subcellular). | Check cellular boundaries and counterstain, compare a positive tissue control, and report the assay-specific discrepancy rather than assigning a mitochondrial pattern (general IHC practice). |
| Adipocytes or cervical squamous epithelial cells stain strongly. | These populations are recorded as not detected; nonspecific primary binding or endogenous detection activity are possibilities to test (HPA: tissue IHC; general IHC practice). | Compare omission and detection controls, then check whether granular staining remains restricted to the expected cells (general IHC practice). |
| Only scattered cells stain in a nominally positive tissue. | HPA's levels refer to named cell populations, so a section with few of those cells may yield little interpretable signal (HPA: tissue IHC; general IHC practice). | Identify the relevant cells on the counterstained section and score their signal separately from surrounding unstained populations (general IHC practice). |
| A second run gives a different apparent intensity. | Run conditions and chromogen development can alter apparent staining strength (general IHC practice); HPA's medium level is a reference observation, not a quantitative threshold (HPA: tissue IHC). | Compare sections processed together with the same control tissue and score compartment, cell type, and background alongside intensity (general IHC practice). |
Experiment evidence
IHC & IF images showing BRI3BP staining
A30536 · IHCImmunohistochemistry analysis of paraffin-embedded human brain tissue, using BRI3B Antibody. The picture on the right is blocked with the sy
HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000184992-BRI3BP/tissue/cerebral+cortex#imid_3850064
HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000184992-BRI3BP/tissue/colon#imid_3849998
HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000184992-BRI3BP/tissue/liver#imid_3849991
HPA · KidneyHuman Protein Atlas IHC, Kidney. Source: https://www.proteinatlas.org/ENSG00000184992-BRI3BP/tissue/kidney#imid_3849992
HPA · ICC-IF RT-4Human Protein Atlas immunofluorescence, RT-4, antibody HPA014957; green = BRI3BP, blue = nucleus, red = microtubules.
HPA · ICC-IF RT-4Human Protein Atlas immunofluorescence, RT-4, antibody HPA014957; green = BRI3BP, blue = nucleus, red = microtubules.
← swipe / scroll for more images →
Real IHC / IF data behind the staining-pattern expectations in this section.
Sample controls for BRI3BP IHC & IF
🧪Run caudate first and score glial cells for staining (HPA: Medium in caudate glial cells). Use adipose tissue as the negative tissue (HPA: Not detected in adipocytes); on the caudate slide, cells without specific staining should show only background chromogen, without assuming a particular cell type is negative.
Positive control tissue: Adrenal gland (Glandular cells, HPA Medium)
Negative control tissue: Adipose tissue (HPA Not detected)
ICC-IF cell lines (HPA subcellular resource): HPA ICC-IF images show BRI3BP in RT-4, with annotated localisation: Nucleoplasm (approved), Mitochondria (approved) (HPA subcellular).
Technical controls: Include a no-primary secondary-only control and a nonimmune antibody matched to the primary antibody’s host species, immunoglobulin class, and clonality. Use a BRI3BP knockout specimen or peptide competition as a specificity control (A30536 caption: peptide-blocked human brain section); for caudate, quench endogenous peroxidase and distinguish granular lipofuscin pigment from chromogen (standard chromogenic IHC practice).
⚠️Feasibility: No supplied target or application evidence reports a BRI3BP-specific fixation window, fixation effect, or antigen-retrieval dependency; optimize retrieval empirically. The selected A30536 caption documents staining in a paraffin-embedded human brain section but does not report the fixative (A30536 caption: fixative unreported). Frozen sections and tissue IF have no supplied comparison showing easier detection; brain lipofuscin may complicate interpretation of granular staining or fluorescence (standard IHC/IF practice).
HPA tissue IHC evidence for BRI3BP
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 |
|---|---|---|---|---|
| Adrenal gland | Glandular cells | Medium | Protein (IHC) | HPA → |
| Appendix | Glandular cells | Medium | Protein (IHC) | HPA → |
| Bone marrow | Hematopoietic cells | Medium | Protein (IHC) | HPA → |
| Breast | Glandular cells | Medium | Protein (IHC) | HPA → |
| Caudate | Glial cells | Medium | Protein (IHC) | HPA → |
Undetected expression · recommended negative controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| Adipose tissue | Adipocytes | Not detected | Protein (IHC) | HPA → |
| Cervix | Squamous epithelial cells | Not detected | Protein (IHC) | HPA → |
| Smooth muscle | Smooth muscle cells | Not detected | Protein (IHC) | HPA → |
| Soft tissue | Fibroblasts | Not detected | Protein (IHC) | HPA → |
| Vagina | Squamous epithelial cells | Not detected | Protein (IHC) | HPA → |
Section 3
Advanced BRI3BP IHC Tips
Troubleshoot BRI3BP chromogenic IHC with matched processing controls and compartment-aware interpretation (UniProt Q8WY22; HPA tissue IHC).
How should I optimize antigen retrieval when BRI3BP staining is weak in paraffin sections?
For paraffin sections, start with citrate pH 6.0 HIER at 95–98 °C for 20 min (page retrieval setting). Cool sections in retrieval buffer before washing, and process comparison sections together so differences reflect the tested condition (standard IHC practice). BRI3BP has four annotated transmembrane segments, so epitope accessibility may depend on processing and antibody epitope location (UniProt Q8WY22). If staining remains weak, compare a modestly longer retrieval time on adjacent sections while monitoring tissue integrity (standard IHC practice). Include the peptide-blocked brain image as evidence for the selected antibody, while recognizing that its caption reports no retrieval conditions (A30536 caption).
Could fixation explain inconsistent BRI3BP staining between paraffin blocks?
Target-specific fixation sensitivity is unknown because the supplied BRI3BP evidence does not report a tested fixative or fixation interval (A30536 caption). The selected brain image identifies paraffin-embedded tissue, but its caption does not establish whether formalin was used (A30536 caption). Document each block’s fixative, fixation duration, processing schedule, and section age before comparing stain intensity (standard IHC practice). For an optimization experiment, stain adjacent sections from blocks with known processing histories in the same run and hold retrieval and detection constant (standard IHC practice). Treat a processing-associated difference as an assay observation until a controlled comparison establishes its cause (standard IHC practice).
Which staining compartments should I expect for BRI3BP in tissue sections?
Expect predominantly granular cytoplasmic tissue staining, the pattern reported across most tissues in the tissue IHC profile (HPA tissue IHC). BRI3BP is annotated at the mitochondrial outer membrane, consistent with a punctate cytoplasmic contribution (UniProt Q8WY22). An independent cell-based localization record also reports nucleoplasm and mitochondria, so nuclear signal warrants evaluation rather than automatic rejection (HPA subcellular). Compare candidate compartments with matched controls and cellular morphology; chromogenic resolution alone cannot establish mitochondrial colocalization (standard IHC practice). Diffuse nuclear-only staining that varies with section edges or background controls deserves additional validation before it is scored as BRI3BP (standard IHC practice; HPA subcellular).
How can epitope placement affect interpretation of BRI3BP IHC?
BRI3BP has 251 amino acids, four annotated transmembrane segments, and no annotated isoforms in the supplied record (UniProt Q8WY22). Its annotated modified residues include acetyllysine at position 229 and phosphoserine at position 248 (UniProt Q8WY22). Without an antibody epitope map, neither a specific membrane-facing epitope nor sensitivity to either modification can be assigned to the catalog antibody (A30536 caption; UniProt Q8WY22). If two antibodies disagree, compare their stated immunogens, retrieval conditions, and peptide-blocking behavior before attributing the discrepancy to protein biology (standard IHC practice). The selected brain caption documents peptide blocking for its own antibody and image only (A30536 caption).
How should I follow up an IHC pattern using multiplex BRI3BP immunofluorescence?
Use IF as a separate validation experiment and pair BRI3BP with a marker for the cell population being scored in the IHC section (standard IF practice). For brain tissue, the IHC profile reports staining in glial cells and cerebellar granular-layer cells, which can guide marker selection without proving coexpression (HPA tissue IHC). Choose fluorophores after checking unstained tissue autofluorescence and single-color controls, favoring channels with a clear signal-to-background separation (standard IF practice). BRI3BP spans membranes four times; choose permeabilization according to the antibody’s documented epitope side, or compare mild permeabilization with an unpermeabilized control if that side is unknown (UniProt Q8WY22; standard IF practice). Assess mitochondrial and nucleoplasmic patterns separately (HPA subcellular).
How can I reduce diffuse or patchy background in BRI3BP chromogenic IHC?
Run a no-primary control and assess endogenous pigment and enzyme activity before attributing diffuse color to BRI3BP (standard IHC practice). Include a peroxidase block for enzyme-based chromogenic detection, and limit DAB development using the same timing across comparison sections (standard IHC practice). Titrate the primary antibody and strengthen blocking or washing when background rises alongside specific staining; no tested dilution is supplied here (standard IHC practice; A30536 caption). Examine section folds, edges, and necrotic regions separately because they can produce misleading intensity patterns (standard IHC practice). Compare the remaining signal with the reported granular cytoplasmic tissue profile and the selected antibody’s peptide-blocked brain image (HPA tissue IHC; A30536 caption).
What is a defensible way to quantify BRI3BP staining across tissue samples? ⚠ ANSWER MARKED FOR VERIFICATION
Define the cell population and scoring compartment before measurement, using granular cytoplasmic staining as the primary tissue IHC pattern (HPA tissue IHC). Report an H-score or both percent positive cells and staining intensity; use positive-cell density per mm² when cell counts vary across sampled areas (standard IHC practice). Normalize counts to viable tissue area or to the number of eligible cells, and apply one threshold and acquisition setting across the comparison set (standard IHC practice). Exclude folds, necrosis, and edge artifacts by a prespecified rule, with the excluded area recorded (standard IHC practice). Score nuclear signal separately because nucleoplasmic localization is also reported in cell-based data (HPA subcellular).
When should an apparent BRI3BP-positive cell be treated as an artifact?
A credible tissue result should fit the reported granular cytoplasmic pattern and the identity of the scored cell population (HPA tissue IHC). Medium staining is reported in brain glial cells and several glandular populations, whereas adipocytes and smooth-muscle cells are listed as not detected in the cited tissue profile (HPA tissue IHC). Isolated edge staining, necrotic deposits, or color persisting in a no-primary control favors a technical source (standard IHC practice). Nuclear staining needs context because nucleoplasmic localization is approved in cell-based imaging, while the broad tissue pattern is granular cytoplasmic (HPA subcellular; HPA tissue IHC). The selected brain peptide-blocking image supports that antibody’s illustrated signal, but does not validate every tissue or compartment (A30536 caption).
Boster reagents
Best BRI3BP / BRI3-binding protein IHC Antibodies
A30536 has an IHC image of paraffin-embedded human brain with a peptide-blocking control (A30536 image caption). IF/ICC and Human/Mouse reactivity are listed; no IF image is supplied (catalog A30536).
Real IHC data
Anti-BRI3B BRI3BP Antibody
Cat # A30536
A30536 will render with IHC data from paraffin-embedded human brain and a peptide-blocked comparison (A30536 image caption). It lists IF/ICC and Human/Mouse reactivity, but supplies no IF image or mouse tissue image (catalog A30536 applications, reactivity, image alts).
Which to pick: Choose A30536 for paraffin-section tissue IHC because its own image documents human brain staining with a peptide-blocking control (A30536 image caption). For IF/ICC or mouse samples, A30536 is the listed polyclonal option based on its applications and reactivity; no IF image or mouse-section example is supplied (catalog A30536 applications, reactivity, dilution_raw, image alts). The IHC caption does not report the fixative (A30536 image caption).
Each figure is that product's own IHC / IF validation image from its datasheet.
References
- UniProt Consortium. UniProt entry Q8WY22 (BRI3B_HUMAN, BRI3-binding protein).
- Human Protein Atlas. BRI3BP tissue IHC expression (reliability: Approved).
- Human Protein Atlas. BRI3BP subcellular location (ICC-IF): Localized to the nucleoplasm and mitochondria..
- Human Protein Atlas. BRI3BP antibody validation summary (1 antibodies).
- The clinicopathological significance of BRI3BP in women with invasive breast cancer. Translational cancer research 2024 — PMC11730447.
- Multi-omic analysis reveals elevated BRI3BP expression associated with hepatocellular carcinoma progression and poor prognosis. Scientific reports 2025 — PMC12578974.
- PubMed PMID:11860200 — UniProt-cited evidence.
- PubMed PMID:16541075 — UniProt-cited evidence.
- PubMed PMID:15489334 — UniProt-cited evidence.
Immunohistochemistry protocol sheet for BRI3BP (BRI3-binding protein). Expected localisation Granular cytoplasm (HPA tissue IHC); mitochondrial outer membrane expected (UniProt). Antigen retrieval: Citrate pH 6.0 HIER, 95–98 °C, 20 min (rule: cytoplasmic / membrane antigen). Validated IHC antibody: A30536 at 1:100 - 1:300 (the product's own IHC validation image is shown). Controls (HPA): positive Adrenal gland (Glandular cells); negative Adipose tissue. IHC-P workflow on paraffin sections; fixative unspecified. Protocol steps: 1. Sample: Paraffin-embedded human brain tissue; fixative not specified (datasheet A30536); 2. Fixation: Image fixative and duration unreported (datasheet A30536); 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-BRI3BP, 1:100 - 1:300 (datasheet A30536); 9. Primary incubation: Overnight at 4 °C (standard). Watch for: Fixation: Keep fixation consistent across paraffin sections. (standard IHC practice; not target-specific); Caveat: Only medium consistency between IHC and RNA (HPA tissue IHC); Isoform / epitope: No annotated isoforms; epitope side across 4 membrane segments is unknown (UniProt); Regulation: Overexpressed in various carcinomas (UniProt). Evidence: UniProt Q8WY22, HPA tissue IHC · Approved, HPA ICC-IF · nucleoplasm and mitochondria., Datasheet A30536, PMC11730447, PMC12578974. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/BRI3BP/immunohistochemistry-ihc-guide.