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BORA / Protein aurora borealis · IHC design guide
Design Immunohistochemistry for BORA
Plan BORA staining in paraffin sections using the cytoplasmic tissue pattern reported by HPA (HPA tissue IHC). Compare candidate positive tissues with adipocytes, where staining was not detected, and start the catalog antibody at 2.5 μg/mL (HPA tissue IHC; datasheet).
Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
BORA 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 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 |
Important caveats
Reasons your staining may differ from the expected pattern.
| Fixation | Keep formalin fixation consistent across sections. (standard IHC practice; not target-specific) | |
| Caveat | Adipocytes may be unstained despite the broad profile (HPA tissue IHC) | |
| Regulation | Expression regulation is unannotated (UniProt) | |
| Isoform / epitope | 2 isoforms; epitope coverage is unspecified (UniProt; datasheet) |
Section 1
Recommended BORA IHC & IF Protocols
The catalog antibody’s IHC-P protocol is accompanied by one published BORA IHC protocol using breast cancer tissue microarray slides (PMC5546444).
| Sample | Tissue sections; selected-image fixative not specified (standard IHC workflow) |
| Fixation | Image fixative and duration unreported (datasheet A02087); 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-BORA, 2.5 μg/mL (datasheet A02087) |
| 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 | BORA-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 retrieval rule); the published protocol used microwave retrieval for 15 min (PMC5546444).
Section 2
What Is the Expected BORA Staining Pattern?
In paraffin-section IHC, expect BORA staining mainly in the cytoplasm across many tissues, including glandular cells, hematopoietic cells and selected other cell populations (HPA tissue IHC: cytoplasmic expression in most tissues; listed High cells). BORA has no annotated transmembrane segment (UniProt Q6PGQ7 topology). Treat the pattern as a comparison point: HPA rates its tissue IHC Approved, with medium staining–RNA consistency and external verification pending (HPA tissue IHC reliability).
| Cytoplasmic staining in adrenal or appendix glandular cells (HPA tissue IHC: High in both). | This matches the reported IHC compartment and cell types (HPA tissue IHC). Compare cells within the section: a High designation identifies a reported positive population, but does not require every cell to stain equally (HPA tissue IHC: High). |
| Predominantly nuclear staining with little cytoplasmic signal in a paraffin section (HPA tissue IHC: cytoplasmic profile). | Investigate a compartment mismatch before scoring it as expected IHC. HPA separately reports nucleoplasm and nucleoli fibrillar-center signal by ICC-IF, with additional cytosol signal; that IF observation does not establish a nuclear IHC pattern (HPA subcellular ICC-IF; HPA tissue IHC). |
| Strong signal in adipocytes while the reported positive cells remain faint (HPA tissue IHC: adipocytes Not detected). | This conflicts with the HPA cell-type comparison and raises concern for nonspecific staining or endogenous detection activity (HPA tissue IHC; general IHC practice). Check the no-primary control and inspect other cells in the same section before assigning BORA positivity (general IHC practice). |
| Broad, fairly uniform color over cells and tissue spaces, without a discernible cytoplasmic pattern (HPA tissue IHC: cytoplasmic profile). | Treat this as background until controls clarify it; diffuse chromogen alone does not identify BORA-positive cells (general IHC practice). Review blocking, washing and detection controls, then reassess whether staining follows the reported cellular compartment (general IHC practice; HPA tissue IHC). |
| No detectable signal in adrenal or appendix glandular cells on the test run (HPA tissue IHC: High in both). | A negative result in a reported High population calls for a technical check before interpreting the study tissue as negative (HPA tissue IHC; general IHC practice). Review the antibody's IHC-P instructions, retrieval, detection and a positive control processed in the same run (general IHC practice). |
💡Expected BORA appearanceCall a result consistent when defined cell populations show cytoplasmic chromogen, including reported High glandular or hematopoietic cells; isolated adipocyte staining or widespread structureless color is suspect (HPA tissue IHC: cytoplasmic profile, High populations, adipocytes Not detected; general IHC practice).
| Tissue and cell selection (HPA tissue IHC). | HPA reports High staining in adrenal, appendix, duodenal and endometrial glandular cells; bone-marrow hematopoietic cells; cerebellar granular-layer cells; colonic endothelial cells; and esophageal squamous epithelial cells (HPA tissue IHC). Adipocytes are Not detected, while caudate glial and ovarian follicle cells are Low (HPA tissue IHC). Select controls by cell type within the tissue (general IHC practice). |
| Compartment by application (HPA tissue IHC; HPA subcellular ICC-IF). | IHC has a cytoplasmic tissue profile, whereas ICC-IF reports approved nucleoplasm and nucleoli fibrillar-center locations plus cytosol (HPA tissue IHC; HPA subcellular ICC-IF). IF/ICC Q&A: What should IF show? Those reported nuclear locations and additional cytosol; the separate IF/ICC guide should govern its workflow (HPA subcellular ICC-IF). |
| Evidence and antibody validation (HPA tissue IHC; HPA antibodies). | The supplied HPA antibody HPA040866 is Approved for IHC and ICC; the tissue profile has medium staining–RNA consistency and awaits external verification (HPA antibodies; HPA tissue IHC reliability). Use concordant controls to support a local interpretation without treating approval as proof that every stained cell is specific (general IHC practice). |
| Topology, processing and variants (UniProt Q6PGQ7). | BORA has no annotated transmembrane segment, signal peptide or propeptide, and UniProt lists two isoforms (UniProt Q6PGQ7 topology, processing, isoforms). Those entries give no basis for expecting a membrane rim or secreted deposit. Isoform recognition by the staining antibody is unspecified in the supplied evidence (UniProt Q6PGQ7; HPA antibodies). |
| Phosphorylation and target-specific retrieval (UniProt Q6PGQ7). | UniProt lists six modified residues, including phosphoserines, but the supplied evidence gives no antibody epitope or BORA-specific retrieval response (UniProt Q6PGQ7 modified residues; HPA antibodies). Do not infer that phosphorylation or fixation explains a weak or absent stain; optimize retrieval using the IHC antibody instructions and controls (general IHC practice). |
| Situation | Likely cause | Next action |
|---|---|---|
| Positive-control glandular cells are blank (HPA tissue IHC: adrenal and appendix High). | The run may have failed at retrieval, primary-antibody incubation or detection (general IHC practice). | Check the IHC-P antibody instructions and reagent sequence; repeat with a reported High tissue and a same-run detection control (general IHC practice; HPA tissue IHC). |
| Every compartment appears faintly brown, including spaces between cells (general IHC practice). | Diffuse background can obscure the reported cytoplasmic pattern (general IHC practice; HPA tissue IHC). | Inspect a no-primary control; review blocking, wash steps and chromogen development, then rescore identifiable cells (general IHC practice). |
| Adipocytes stain as strongly as reported High cells (HPA tissue IHC: adipocytes Not detected). | Nonspecific binding or endogenous detection activity is possible (general IHC practice). | Compare no-primary and detection controls, then reassess staining in the named cell populations (general IHC practice; HPA tissue IHC). |
| Only nuclei stain in IHC-P sections (HPA tissue IHC: cytoplasmic profile). | The result disagrees with the tissue IHC profile; nuclear ICC-IF localization alone cannot validate it as IHC (HPA tissue IHC; HPA subcellular ICC-IF). | Check the positive-control compartment, no-primary control and morphology before reporting nuclear IHC positivity (general IHC practice). |
| A Low population is scored negative despite a working positive control (HPA tissue IHC: caudate glia and ovarian follicle cells Low). | Weak reported staining may fall below the run's visual detection threshold (HPA tissue IHC; general IHC practice). | Record the detection limit and avoid using a Low population alone to judge run failure (general IHC practice; HPA tissue IHC). |
| Strong staining appears outside the reported cell population in an otherwise positive tissue (HPA tissue IHC: cell-specific entries). | The tissue name alone does not establish that each stained cell expresses BORA (HPA tissue IHC; general IHC practice). | Identify cells by morphology, compare the HPA cell-type entry, and check no-primary staining before scoring them (HPA tissue IHC; general IHC practice). |
Experiment evidence
IHC & IF images showing BORA staining
A02087 · IHCImmunohistochemistry of Bora in mouse brain tissue with Bora antibody at 2.5 μg/mL.
A02087 · IFImmunofluorescence of Bora in Mouse Brain cells with Bora antibody at 20 μg/mL.
HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000136122-BORA/tissue/cerebral+cortex#imid_10673137
HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000136122-BORA/tissue/colon#imid_10673063
HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000136122-BORA/tissue/liver#imid_10673056
HPA · KidneyHuman Protein Atlas IHC, Kidney. Source: https://www.proteinatlas.org/ENSG00000136122-BORA/tissue/kidney#imid_10673057
HPA · ICC-IF A-431Human Protein Atlas immunofluorescence, A-431, antibody HPA040866; green = BORA, blue = nucleus, red = microtubules.
HPA · ICC-IF A-431Human Protein Atlas immunofluorescence, A-431, antibody HPA040866; green = BORA, blue = nucleus, red = microtubules.
← swipe / scroll for more images →
Real IHC / IF data behind the staining-pattern expectations in this section.
Sample controls for BORA IHC & IF
🧪Run adrenal gland first and score its glandular cells, which HPA rates High for BORA (HPA: adrenal gland, glandular cells, High). Run adipose tissue as the negative comparator because adipocytes are Not detected (HPA: adipose tissue, adipocytes, Not detected); on the adrenal slide, use adjacent non-glandular cells as a background comparator without assuming they are BORA-negative.
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 BORA in A-431, U-251MG, U2OS, with annotated localisation: Nucleoplasm (approved), Nucleoli fibrillar center (approved) (HPA subcellular).
Technical controls: Include a no-primary (secondary-only) control, an isotype control matched to the primary antibody’s host species and immunoglobulin class, and BORA-knockout material as a biological negative. For chromogenic staining of adrenal gland, block endogenous peroxidase and check blood-rich areas for background before interpreting glandular staining (HPA: adrenal gland, glandular cells, High).
⚠️Feasibility: A target-specific fixation window or fixation effect is unreported, and the selected A02087 mouse-brain IHC caption does not state a fixative (A02087 tissue-IHC caption: fixative not stated). Retrieval dependency is unreported; optimize antigen retrieval for paraffin sections before scoring. Frozen-section ease is unreported, while ICC-IF images exist for A-431, U-251MG and U2OS (HPA: subcellular ICC-IF images); in adrenal sections, distinguish glandular signal from endogenous peroxidase background.
HPA tissue IHC evidence for BORA
Comprehensive Human Protein Atlas IHC scoring per tissue (reliability: Approved — Medium consistency between antibody staining and RNA expression data. 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 |
|---|---|---|---|---|
| Adrenal gland | Glandular cells | High | Protein (IHC) | HPA → |
| Appendix | Glandular cells | High | Protein (IHC) | HPA → |
| Bone marrow | Hematopoietic cells | High | Protein (IHC) | HPA → |
| Cerebellum | Cells in granular layer | High | Protein (IHC) | HPA → |
| Colon | Endothelial cells | High | Protein (IHC) | HPA → |
Undetected expression · recommended negative controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| Adipose tissue | Adipocytes | Not detected | Protein (IHC) | HPA → |
Section 3
Advanced BORA IHC Tips
Use compartment, cell type, and matched controls to troubleshoot BORA staining in chromogenic tissue IHC (HPA tissue IHC; HPA subcellular).
Which retrieval conditions should I try first for weak BORA staining?
Start with citrate buffer at pH 6.0, heated to 95–98 °C for 20 min (page retrieval setting). Cool sections consistently before antibody incubation, and compare retrieved with untreated sections from the same tissue block (standard IHC practice). If staining remains weak, vary heating time in small steps while watching for tissue damage; record each condition alongside its negative control (standard IHC practice). The catalog antibody stained mouse brain tissue at 2.5 µg/mL, but its image caption does not state fixation or retrieval conditions (A02087 caption). Do not treat that image as evidence that a different retrieval buffer was tested (A02087 caption).
How should I troubleshoot BORA staining after fixation?
BORA-specific sensitivity to fixative type or fixation duration is unknown from the supplied evidence (A02087 caption; UniProt Q6PGQ7). The catalog image shows staining in mouse brain at 2.5 µg/mL, but its caption gives no fixative, so it cannot establish a fixation window (A02087 caption). For paraffin IHC, document fixation time and compare sections processed with the same retrieval and antibody conditions before attributing signal differences to BORA biology (standard IHC practice). Include a control section from the same processing run and inspect nuclear detail and tissue integrity (standard IHC practice). If fixation histories differ, report that limitation when comparing staining intensity (standard IHC practice).
Should BORA stain the cytoplasm or nucleus in tissue sections?
Assess cytoplasmic staining first in tissue IHC: HPA reports cytoplasmic expression in most tissues (HPA tissue IHC). Nuclear signal also merits evaluation because HPA cell imaging places BORA mainly in the nucleoplasm and nucleoli fibrillar center, with additional cytosolic localisation (HPA subcellular). UniProt supplies no subcellular annotation for BORA, so compartment agreement cannot be established from that record (UniProt Q6PGQ7). Score nuclear and cytoplasmic staining separately in matched fields, using the same counterstain and exposure or viewing conditions (standard IHC practice). Treat diffuse staining across damaged tissue cautiously, especially when compartment boundaries are no longer discernible (standard IHC practice).
Could isoforms or phosphorylation explain inconsistent BORA staining?
BORA has 2 annotated isoforms, but the supplied evidence does not locate the catalog antibody epitope within either sequence (UniProt Q6PGQ7; A02087 caption). The record also lists phosphorylated serines, including positions 183, 191, 270, 325, and 331 (UniProt Q6PGQ7). Without an epitope map, do not assign a staining difference to splice form or phosphorylation state (UniProt Q6PGQ7; A02087 caption). Compare serial sections with an independent antibody recognizing a documented epitope, if available, and keep retrieval conditions matched (standard IHC practice). Describe any resulting discrepancy as antibody-dependent until further specificity evidence resolves it (standard IHC practice).
How can IF help check BORA localisation seen by chromogenic IHC?
For a separate IF experiment, multiplex BORA with a validated marker for the expected cell population, such as cerebellar granular-layer cells when examining cerebellum (HPA tissue IHC; standard IF practice). Choose spectrally separated fluorophores and inspect unstained tissue first, assigning the brighter channel where tissue autofluorescence is greatest (standard IF practice). BORA has no annotated transmembrane segment, while HPA reports nuclear and cytosolic signal; use controlled permeabilisation to expose these intracellular compartments (UniProt Q6PGQ7; HPA subcellular; standard IF practice). Compare single-stain controls before interpreting colocalisation (standard IF practice). Keep IF findings distinct from the catalog mouse-brain IHC image, whose caption supplies no IF conditions (A02087 caption).
What should I check when BORA IHC has widespread background?
Check a no-primary control and inspect whether color persists in tissue compartments or along section edges (standard IHC practice). In a peroxidase and DAB workflow, block endogenous peroxidase and compare the control with the fully stained section before calling weak diffuse color BORA-positive (standard IHC practice). Titrate the primary antibody around the reported 2.5 µg/mL mouse-brain image concentration while keeping retrieval, incubation, and detection constant; that caption does not establish an optimal working concentration for other sections (A02087 caption; standard IHC practice). Confirm blocking and washing are consistent across slides (standard IHC practice). Evaluate discrete cellular staining separately from pigment, precipitate, folds, and damaged edges (standard IHC practice).
How should I quantify BORA staining across tissue sections? ⚠ ANSWER MARKED FOR VERIFICATION
Define the cell population and region before scoring, then report the percentage of positive cells and staining intensity separately (standard IHC practice). For an H-score, multiply the percentage at each intensity by its 0–3 category and sum to a 0–300 score (standard IHC practice). Score nuclear and cytoplasmic compartments separately because tissue IHC and cell imaging report different predominant patterns (HPA tissue IHC; HPA subcellular). Normalize positive-cell counts to all evaluable cells in the region, or area-based counts to mm² of viable tissue (standard IHC practice). Apply one threshold across slides and record excluded folds, necrosis, and edge regions (standard IHC practice).
How can I distinguish true BORA signal from staining artefacts?
Look for reproducible cellular staining in viable tissue with a clear compartment and a clean no-primary control (standard IHC practice). HPA reports cytoplasmic tissue staining, including high staining in cerebellar granular-layer cells, while its cell imaging also supports nuclear and cytosolic localisation; interpret either compartment with the assay context stated (HPA tissue IHC; HPA subcellular). Be cautious when apparent positives occur only at cut edges, in necrotic areas, or where endogenous peroxidase also colors the control (standard IHC practice). HPA lists adipocytes as not detected, offering a contextual comparison rather than an absolute negative control for every preparation (HPA tissue IHC). Corroborate unexpected patterns with independently validated specificity controls (standard IHC practice).
Boster reagents
Best BORA / Protein aurora borealis IHC Antibodies
A02087 has IHC data in mouse brain tissue and IF data in mouse brain cells (catalog image captions); Human, Mouse and Rat reactivity is listed (catalog reactivity).
Real IHC data
Anti-Protein aurora borealis Bora Antibody
Cat # A02087
A02087 will render with an IHC image of mouse brain tissue at 2.5 μg/mL (IHC image caption). The same SKU has an IF image of mouse brain cells at 20 μg/mL (IF image caption).
Which to pick: Choose A02087 for tissue IHC: its mouse brain tissue image uses 2.5 μg/mL (IHC image caption), and IHC-P is listed (catalog applications); the fixative is unreported (IHC image caption). For IF/ICC, A02087 has an IF image in mouse brain cells at 20 μg/mL (IF image caption). For other species, A02087 lists Human, Mouse and Rat reactivity (catalog reactivity), while the supplied IHC and IF images show mouse samples (catalog image captions).
Each figure is that product's own IHC / IF validation image from its datasheet.
References
- UniProt Consortium. UniProt entry Q6PGQ7 (BORA_HUMAN, Protein aurora borealis).
- Human Protein Atlas. BORA tissue IHC expression (reliability: Approved).
- Human Protein Atlas. BORA subcellular location (ICC-IF): Mainly localized to the nucleoplasm and nucleoli fibrillar center. In addition localized to the cytosol..
- Human Protein Atlas. BORA antibody validation summary (1 antibodies).
- Cell cycle protein Bora serves as a novel poor prognostic factor in multiple adenocarcinomas. Oncotarget 2017 — PMC5546444.
- Aurora Borealis (Bora), Which Promotes Plk1 Activation by Aurora A, Has an Oncogenic Role in Ovarian Cancer. Cancers 2020 — PMC7226261.
- PubMed PMID:14702039 — UniProt-cited evidence.
- PubMed PMID:15057823 — UniProt-cited evidence.
- PubMed PMID:15489334 — UniProt-cited evidence.
Immunohistochemistry protocol sheet for BORA (Protein aurora borealis). 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: A02087 at 2.5 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 A02087); 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-BORA, 2.5 μg/mL (datasheet A02087); 9. Primary incubation: Overnight at 4 °C (standard). Watch for: Fixation: Keep formalin fixation consistent across sections. (standard IHC practice; not target-specific); Caveat: Adipocytes may be unstained despite the broad profile (HPA tissue IHC); Isoform / epitope: 2 isoforms; epitope coverage is unspecified (UniProt; datasheet); Regulation: Expression regulation is unannotated (UniProt). Evidence: UniProt Q6PGQ7, HPA tissue IHC · Approved, HPA ICC-IF · Mainly localized to the nucleoplasm and nucleoli fibrillar center.…, Datasheet A02087, PMC5546444. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/BORA/immunohistochemistry-ihc-guide.