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BIRC6 / Dual E2 ubiquitin-conjugating enzyme/E3 ubiquitin-protein ligase BIRC6 · IHC design guide
Design Immunohistochemistry for BIRC6
Plan chromogenic BIRC6 IHC in paraffin sections around the cytoplasmic tissue pattern (HPA tissue IHC). Compare high-staining colon glandular cells with squamous epithelial cells reported as undetected in esophagus, while accounting for medium consistency between antibody staining and RNA expression (HPA tissue IHC).
Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
BIRC6 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 most tissues (HPA tissue IHC) | |
| Staining pattern | Cytoplasmic staining across many tissue cell types (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 sections. (standard IHC practice; not target-specific) | |
| Caveat | Antibody staining and RNA show medium consistency (HPA tissue IHC) | |
| Regulation | Staining varies by cell type (HPA tissue IHC) | |
| Isoform / epitope | No annotated isoforms; one chain with no transmembrane segment (UniProt) |
Section 1
Recommended BIRC6 IHC & IF Protocols
Compare the catalog antibody’s IHC-P protocol (datasheet) with four published BIRC6 IHC protocols (PMC10706016; PMC4196171; PMC4416929; PMC7317768).
| Sample | Tissue sections; selected-image fixative not specified (standard IHC workflow) |
| Fixation | Image fixative and duration unreported (datasheet A01773); 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-BIRC6, 5 μg/mL (datasheet A01773) |
| 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 | BIRC6-positive staining in glandular cells of adrenal gland (HPA tissue IHC: High). HPA tissue profile: Cytoplasmic expression in most tissues. 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); optimize for the tissue and antibody.
Section 2
What Is the Expected BIRC6 Staining Pattern?
BIRC6 staining is expected mainly in the cytoplasm of many tissue cell types, with high staining reported in selected glandular, respiratory epithelial, and trophoblastic cells (HPA: tissue IHC). Its Golgi and endosomal associations and cell cycle dependent redistribution can shape localisation (UniProt Q9NR09: subcellular location). BIRC6 has no transmembrane segment (UniProt Q9NR09: topology). HPA rates tissue IHC reliability Approved, with medium consistency between staining and RNA data (HPA: tissue IHC).
| Cytoplasmic staining in colon glandular cells or bronchial respiratory epithelial cells. | This matches reported high staining in those cell types (HPA: tissue IHC). Compare cells within the section; high describes the HPA observation, not a required intensity for every run (HPA: tissue IHC). |
| A sharply nuclear dominant pattern, with little cytoplasmic staining. | Treat this as discordant with the reported cytoplasmic tissue pattern (HPA: tissue IHC) and Golgi, endosomal, and cytoskeletal locations (UniProt Q9NR09: subcellular location). Check whether counterstain or nonspecific detection is being mistaken for target signal (general IHC practice). |
| Strong staining confined to adipocytes, while expected positive glandular cells are unstained. | Adipocytes are reported as not detected, whereas colon glandular cells are high (HPA: tissue IHC). Consider cross-reactivity or endogenous chromogen generating activity, but first verify tissue identity and assay controls (general IHC practice). |
| Uniform haze over cells, stroma, and empty areas. | A field wide deposit is difficult to reconcile with the reported cell associated cytoplasmic pattern (HPA: tissue IHC). It suggests nonspecific background or detection reagent deposition; assess a no primary control (general IHC practice). |
| No staining in a section expected to contain high expressing cells. | Absence in colon glandular cells or placental trophoblastic cells conflicts with their reported high staining (HPA: tissue IHC). Check section quality, retrieval, antibody application, and detection before concluding that the sample lacks BIRC6 (general IHC practice). |
💡Expected BIRC6 appearanceCall a positive result when cytoplasmic signal is clear in expected glandular or respiratory epithelial cells, with high staining possible in the HPA listed tissues; nuclear dominant staining or uniform haze is suspect (HPA: tissue IHC; general IHC practice).
| Which tissue and cell type should anchor interpretation? | Use a reported high group such as colon glandular cells or bronchial respiratory epithelial cells (HPA: tissue IHC). A negative comparator must match the specified cell type: adipocytes, not every cell in adipose tissue, are listed as not detected (HPA: tissue IHC). |
| How much confidence does tissue IHC validation provide? | Tissue staining is Approved but has medium consistency with RNA data (HPA: tissue IHC). HPA074409 is IHC Approved; HPA074738 has ICC approval but no IHC status in the supplied record (HPA: antibodies). Treat unexpected patterns as findings to verify, not proof of a new localisation. |
| Does subcellular location vary through division? | BIRC6 is associated with a pericentriolar compartment in interphase, spindle poles in metaphase, and the midbody ring during cytokinesis (UniProt Q9NR09: subcellular location). A rare division associated focus may therefore be compatible with the record; score the prevalent cytoplasmic pattern separately. |
| Does membrane topology imply surface staining? | No transmembrane segment is reported (UniProt Q9NR09: topology). Its trans Golgi network membrane and endosomal localisation does not establish plasma membrane staining as the expected tissue IHC pattern (UniProt Q9NR09: subcellular location; HPA: tissue IHC). |
| IF/ICC: what pattern should appear? | Golgi apparatus and vesicles are the supported main locations, with an additional supported mitotic spindle location (HPA: subcellular ICC-IF). These resolved IF structures provide context for localisation; routine chromogenic sections are assessed against the broader cytoplasmic tissue pattern (HPA: tissue IHC). |
| What is known about antigen retrieval sensitivity? | No target specific retrieval or fixation effect is established by the supplied UniProt and HPA records. Retrieval is a general paraffin IHC optimisation variable; compare conditions with tissue and reagent controls before attributing a weak result to BIRC6 epitope masking (general IHC practice). |
| Situation | Likely cause | Next action |
|---|---|---|
| A reported high cell type is blank. | The result conflicts with the reported high staining in colon glandular cells or placental trophoblastic cells (HPA: tissue IHC); assay failure is one possibility (general IHC practice). | Confirm the intended cells are present, then review primary antibody use, retrieval, chromogen development, and a concurrently stained positive control (general IHC practice). |
| All sections show uniform brown background. | Diffuse deposition is inconsistent with the reported cytoplasmic cell pattern (HPA: tissue IHC); nonspecific binding or detection background may contribute (general IHC practice). | Inspect a no primary control, blocking and washes, and the detection reagents. Adjust those general IHC conditions before scoring cellular staining (general IHC practice). |
| Signal appears only where endogenous detection activity is likely. | Endogenous activity can create chromogenic signal independent of primary antibody binding (general IHC practice). Such signal cannot by itself establish BIRC6 expression. | Run a no primary control and apply the detection system's appropriate endogenous activity block; then reassess cell specific cytoplasmic staining (general IHC practice). |
| Staining is nuclear dominant. | That pattern differs from HPA's cytoplasmic tissue profile (HPA: tissue IHC) and the supplied UniProt locations (UniProt Q9NR09: subcellular location). | Check counterstain and no primary controls, then compare with a reported high cytoplasmic cell type before accepting the signal as BIRC6 (HPA: tissue IHC; general IHC practice). |
| A low staining tissue is used as the sole negative control. | Thyroid glandular cells and duodenal glandular cells are listed as low, while BIRC6 has low tissue specificity overall (HPA: tissue IHC). Weak signal there need not indicate assay background. | Use a cell type reported as not detected, such as adipocytes, and retain a no primary control to distinguish tissue signal from reagent background (HPA: tissue IHC; general IHC practice). |
| An ICC approved antibody is assumed suitable for paraffin IHC. | HPA074738 has ICC approval but no IHC status in the supplied antibody record; HPA074409 has IHC approval (HPA: antibodies). | Base paraffin tissue expectations on the IHC approved antibody and the HPA tissue pattern. Do not transfer ICC validation or its resolved Golgi and vesicle appearance into an IHC performance claim (HPA: antibodies; HPA: tissue IHC; HPA: subcellular ICC-IF). |
Experiment evidence
IHC & IF images showing BIRC6 staining
A01773 · IHCImmunohistochemistry of BRUCE in human small intestine tissue with BRUCE antibody at 5 μg/mL.
HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000115760-BIRC6/tissue/cerebral+cortex#imid_21009739
HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000115760-BIRC6/tissue/colon#imid_21012985
HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000115760-BIRC6/tissue/liver#imid_21012994
HPA · KidneyHuman Protein Atlas IHC, Kidney. Source: https://www.proteinatlas.org/ENSG00000115760-BIRC6/tissue/kidney#imid_21012987
HPA · ICC-IF MCF-7Human Protein Atlas immunofluorescence, MCF-7, antibody HPA074409; green = BIRC6, blue = nucleus, red = microtubules.
HPA · ICC-IF MCF-7Human Protein Atlas immunofluorescence, MCF-7, antibody HPA074409; green = BIRC6, blue = nucleus, red = microtubules.
← swipe / scroll for more images →
Real IHC / IF data behind the staining-pattern expectations in this section.
Sample controls for BIRC6 IHC & IF
🧪Run colon first: its glandular cells should stain (HPA: High in colon glandular cells). Use oral mucosa squamous epithelial cells as a negative tissue (HPA: Not detected in oral mucosa squamous epithelial cells); on the colon slide, non-glandular cells should be scored as internal negatives only if they show no specific signal above background.
Positive control tissue: Adrenal gland (Glandular cells, HPA High)
Negative control tissue: Adipose tissue (HPA Not detected)
ICC-IF cell lines (HPA subcellular resource): HPA ICC-IF images show BIRC6 in MCF-7, SH-SY5Y, U2OS, A-431, A-549, with annotated localisation: Golgi apparatus (supported), Vesicles (supported) (HPA subcellular).
Technical controls: Include a no-primary (secondary-only) control and a primary-matched control: host- and isotype-matched antibody for a monoclonal primary, or nonimmune IgG from the same host for a polyclonal primary (standard IHC practice). Confirm specificity with BIRC6 knockout material, or a peptide block if the immunizing peptide is available; quench endogenous peroxidase in colon sections for chromogenic detection (standard IHC practice).
⚠️Feasibility: A target-specific fixation window is unreported, and the fixative is unreported for the A01773 human small intestine IHC caption (selected-SKU tissue-IHC caption). Antigen-retrieval dependence is also unreported; optimize retrieval on matched positive and negative sections (standard IHC practice). Frozen sections have no demonstrated ease advantage in the supplied evidence, while IF/ICC images support assessment of Golgi, vesicular and mitotic spindle localization (HPA: subcellular ICC-IF); endogenous peroxidase from blood cells in colon can cause chromogenic background (standard IHC practice).
HPA tissue IHC evidence for BIRC6
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 | High | Protein (IHC) | HPA → |
| Breast | Glandular cells | High | Protein (IHC) | HPA → |
| Bronchus | Respiratory epithelial cells | High | Protein (IHC) | HPA → |
| Colon | Glandular cells | High | Protein (IHC) | HPA → |
| Fallopian tube | Glandular cells | High | Protein (IHC) | HPA → |
Undetected expression · recommended negative controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| Adipose tissue | Adipocytes | Not detected | Protein (IHC) | HPA → |
| Caudate | Glial cells | Not detected | Protein (IHC) | HPA → |
| Esophagus | Squamous epithelial cells | Not detected | Protein (IHC) | HPA → |
| Lymph node | Non-germinal center cells | Not detected | Protein (IHC) | HPA → |
| Oral mucosa | Squamous epithelial cells | Not detected | Protein (IHC) | HPA → |
Section 3
Advanced BIRC6 IHC Tips
Troubleshoot BIRC6 staining in paraffin sections by checking retrieval, tissue controls, compartment patterns, and scoring before interpreting chromogenic signal.
How should I retrieve BIRC6 in paraffin sections when cytoplasmic staining is weak?
Start with citrate buffer at pH 6.0 for heat-induced epitope retrieval at 95–98 °C for 20 minutes (page retrieval rule). Let sections cool consistently, then compare BIRC6 staining across matched sections while keeping antibody concentration and detection conditions fixed (standard IHC practice). If staining remains weak, test a different retrieval buffer or pH on adjacent sections as a fallback, with an identically processed control (standard IHC practice). Score signal in the expected cytoplasmic region rather than treating any darker section as improved, because tissue staining is chiefly cytoplasmic (HPA tissue IHC).
Could fixation explain weak or uneven BIRC6 staining?
Target-specific sensitivity of BIRC6 to fixation is unknown from the supplied evidence; the selected small-intestine IHC caption does not state its fixative (A01773 caption). For paraffin IHC, compare sections with documented fixation histories before changing retrieval or antibody conditions, since fixation can alter epitope accessibility (standard IHC practice). Examine whether weak signal follows a processing boundary or affects the whole section, and include a concurrently stained reference section (standard IHC practice). Keep the initial citrate pH 6.0, 95–98 °C, 20-minute retrieval constant during that comparison (page retrieval rule). Do not assign a BIRC6-specific fixation effect from its localization or modification record (UniProt Q9NR09).
Which BIRC6 staining pattern should I expect in tissue sections?
Expect predominantly cytoplasmic staining across tissues, while assessing the cell type within each structure (HPA tissue IHC). BIRC6 can occupy the trans-Golgi network and endosomes in interphase, then move toward spindle poles and the midbody during cell division (UniProt Q9NR09 localization). A compact perinuclear pattern or a focal signal in a dividing cell can therefore warrant closer inspection, but chromogenic tissue sections may not resolve those structures reliably (UniProt Q9NR09 localization; standard IHC practice). Compare the pattern with nearby glandular cells, which show high staining in colon, and with adipocytes, where staining is not detected (HPA tissue IHC).
How can I assess an unexpected BIRC6 pattern when the antibody epitope is unspecified?
The supplied record lists 0 annotated isoforms, and the antibody epitope is not specified, so neither explains an unexpected staining pattern by itself (UniProt Q9NR09 isoforms; supplied antibody evidence). BIRC6 is a 4,857-amino-acid protein with a ubiquitin-like region at residues 3819–4068 and a UBC core at 4573–4740 (UniProt Q9NR09). Its listed modifications include phosphoserines at 473 and 480, but their effect on this antibody cannot be inferred without epitope information (UniProt Q9NR09 modified residues; supplied antibody evidence). Check the antibody's documented immunogen or epitope, then compare staining with an independent reagent or orthogonal expression evidence (standard IHC practice).
How should I follow up an IHC BIRC6 pattern with multiplex immunofluorescence?
Use IF as a separate validation experiment, since this IHC guide does not establish an IF protocol for the catalog antibody (supplied application scope). Multiplex BIRC6 with a marker identifying the expected cell type, such as a validated glandular-cell marker when examining colon glands, where glandular BIRC6 staining is high (HPA tissue IHC; standard IF practice). Choose fluorophores whose emission is distinct from the specimen's autofluorescence and include single-stain and no-primary controls (standard IF practice). Permeabilize sufficiently to reach the cytoplasmic epitope, while optimizing detergent exposure because the antibody's epitope and its accessibility are unspecified (HPA tissue IHC; supplied antibody evidence; standard IF practice).
What should I change when BIRC6 staining obscures tissue detail?
First compare a no-primary control and a positive reference section processed with the same detection reagents (standard IHC practice). In chromogenic IHC, block endogenous peroxidase before DAB development and check whether pigment persists without primary antibody (standard IHC practice). Titrate the primary antibody and shorten detection or chromogen development if diffuse color masks cellular boundaries, recording each change against the reference (standard IHC practice). The selected small-intestine image used 5 μg/mL, but its caption does not establish that concentration for every tissue or processing condition (A01773 caption). Judge improvement by preservation of the expected cytoplasmic pattern (HPA tissue IHC).
How should I score BIRC6 staining across tissue samples? ⚠ ANSWER MARKED FOR VERIFICATION
Define the cell population and cytoplasmic compartment before scoring, because BIRC6 tissue staining is chiefly cytoplasmic (HPA tissue IHC). For comparable sections, record the percentage of positive cells and staining intensity, then calculate an H-score using percentages at each intensity level (standard IHC practice). If counting focal stained cells, report density per mm² of evaluable tissue and specify whether dividing cells were included (standard IHC practice; UniProt Q9NR09 localization). Normalize counts to the same eligible cell population or tissue area across samples, excluding folds, necrosis, and section edges by a preset rule (standard IHC practice).
When is an apparent BIRC6-positive cell likely to be an artefact?
Treat a reproducible cytoplasmic signal in the expected cell population as more credible than isolated color in an unrelated compartment (HPA tissue IHC; standard IHC practice). High glandular-cell staining in colon and absent staining in adipocytes offer tissue-level comparison points, though the HPA tissue annotation has medium staining-to-RNA consistency (HPA tissue IHC). Inspect nuclear-only color, section-edge accentuation, necrotic debris, and staining that survives omission of primary antibody before calling a cell positive (standard IHC practice). Perinuclear or division-associated foci need careful morphology because BIRC6 also localizes to Golgi-related structures, spindle poles, and the midbody (UniProt Q9NR09 localization).
Boster reagents
Best BIRC6 / Dual E2 ubiquitin-conjugating enzyme/E3 ubiquitin-protein ligase BIRC6 IHC Antibodies
The IHC-validated BIRC6 antibody has a human small-intestine IHC image (A01773 image caption). IF/ICC data are not listed (catalog: applications).
Real IHC data
Anti-BRUCE BIRC6 Antibody
Cat # A01773
A01773 is listed for human IHC-P (catalog: applications and reactivity). Its own image shows human small-intestine staining at 5 μg/mL (A01773 image caption).
Which to pick: Choose A01773, a rabbit polyclonal antibody, for human paraffin-section IHC (catalog: host, clonality, reactivity, IHC-P; A01773 image caption: human small intestine). Its image caption does not report the fixative (A01773 image caption). No IF/ICC or cross-species option is listed (catalog: A01773 applications and human-only reactivity).
Each figure is that product's own IHC / IF validation image from its datasheet.
References
- UniProt Consortium. UniProt entry Q9NR09 (BIRC6_HUMAN, Dual E2 ubiquitin-conjugating enzyme/E3 ubiquitin-protein ligase BIRC6).
- Human Protein Atlas. BIRC6 tissue IHC expression (reliability: Approved).
- Human Protein Atlas. BIRC6 subcellular location (ICC-IF): Mainly localized to the Golgi apparatus and vesicles. In addition localized to the mitotic spindle..
- Human Protein Atlas. BIRC6 antibody validation summary (2 antibodies).
- Evaluation of BIRC6 Expression in Oral Squamous Cell Carcinoma, Epithelial Dysplasia, Lichen Planus with and without Dysplasia, and Hyperkeratosis. Diagnostics (Basel, Switzerland) 2023 — PMC10706016.
- The BIRC6 gene as a novel target for therapy of prostate cancer: dual targeting of inhibitors of apoptosis. Oncotarget 2014 — PMC4196171.
- Overexpression of BIRC6 Is a Predictor of Prognosis for Colorectal Cancer. PloS one 2015 — PMC4416929.
- Expression of inhibitors of apoptosis proteins in salivary gland adenoid cystic carcinoma: XIAP is an independent marker of impaired cause-specific survival. Clinical otolaryngology : official journal of ENT-UK ; official journal of Netherlands Society for Oto-Rhino-Laryngology & Cervico-Facial Surgery 2020 — PMC7317768.
- PubMed PMID:15815621 — UniProt-cited evidence.
- PubMed PMID:10544019 — UniProt-cited evidence.
- PubMed PMID:10574462 — UniProt-cited evidence.
Immunohistochemistry protocol sheet for BIRC6 (Dual E2 ubiquitin-conjugating enzyme/E3 ubiquitin-protein ligase BIRC6). Expected localisation Cytoplasmic staining in most tissues (HPA tissue IHC). Antigen retrieval: Citrate pH 6.0 HIER, 95–98 °C, 20 min (rule: cytoplasmic / membrane antigen). Validated IHC antibody: A01773 at 5 μg/mL (the product's own IHC validation image is shown). Controls (HPA): positive Adrenal gland (Glandular cells); negative Adipose tissue. IHC workflow; specimen preparation unspecified. Protocol steps: 1. Sample: Tissue sections; selected-image fixative not specified (standard IHC workflow); 2. Fixation: Image fixative and duration unreported (datasheet A01773); 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-BIRC6, 5 μg/mL (datasheet A01773); 9. Primary incubation: Overnight at 4 °C (standard). Watch for: Fixation: Keep fixation consistent across sections. (standard IHC practice; not target-specific); Caveat: Antibody staining and RNA show medium consistency (HPA tissue IHC); Isoform / epitope: No annotated isoforms; one chain with no transmembrane segment (UniProt); Regulation: Staining varies by cell type (HPA tissue IHC). Evidence: UniProt Q9NR09, HPA tissue IHC · Approved, HPA ICC-IF · Mainly localized to the Golgi apparatus and vesicles. In addition…, Datasheet A01773, PMC10706016, PMC4196171. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/BIRC6/immunohistochemistry-ihc-guide.