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CYP7B1 / Cytochrome P450 7B1 · IHC design guide
Design Immunohistochemistry for CYP7B1
Plan paraffin-section CYP7B1 IHC using the IHC-validated antibody at 2–5 μg/ml (datasheet A03545-3). Compare staining with the reported cytoplasmic and nuclear tissue pattern, while accounting for its uncertain reliability (HPA tissue IHC).
Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
CYP7B1 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/nuclear staining (HPA tissue IHC); ER membrane (UniProt) | |
| Staining pattern | Cytoplasmic and nuclear staining in most tissues (HPA tissue IHC) | |
| Antigen retrieval | EDTA pH 8.0 HIER, heat-mediated (datasheet A03545-3) | |
| Positive control | Adrenal gland+4 more · see all | |
| Negative control | None in HPA (detected in all 45 tissues); use no-primary + isotype controls |
Important caveats
Reasons your staining may differ from the expected pattern.
| Fixation | Keep paraffin-section fixation consistent (standard IHC practice; not target-specific) | |
| Caveat | Presumed off-target binding; reliability uncertain (HPA tissue IHC) | |
| Regulation | Regulation not specified (UniProt) | |
| Isoform / epitope | No annotated isoforms; one 1–506 chain (UniProt) |
Section 1
Recommended CYP7B1 IHC & IF Protocols
Compare the catalog antibody’s IHC-P protocol (datasheet A03545-3) with the published rat-testis protocol (PMC6993528).
| Sample | Paraffin-embedded human breast cancer tissue; fixative not specified (datasheet A03545-3) |
| Fixation | Image fixative and duration unreported (datasheet A03545-3); verify before use. |
| Sectioning | 4–5 µm sections on charged slides (standard) |
| Deparaffinisation | Xylene, graded ethanol series to water (standard) |
| Antigen retrieval | Heat retrieval: EDTA pH 8.0 (datasheet A03545-3); 20 min, 95–100 °C (standard) |
| Peroxidase block | 3% H2O2, 10 min, room temperature (standard) |
| Blocking | 10% goat serum (datasheet A03545-3) |
| Primary antibody | Rabbit anti-CYP7B1, 2-5 μg/ml (datasheet A03545-3) |
| Primary incubation | Overnight at 4 °C (datasheet A03545-3) |
| Detection | HRP-conjugated secondary, DAB chromogen (datasheet A03545-3) |
| Counterstain | Hematoxylin, blue, dehydrate and mount (standard) |
| Expected result | CYP7B1-positive staining in glandular cells of adrenal gland (HPA tissue IHC: High). HPA tissue profile: Cytoplasmic and nuclear expression in most tissues. No signal in the no-primary control. |
💡Decision noteStart with heat-mediated EDTA pH 8.0 retrieval for the catalog antibody (datasheet A03545-3); the published protocol used citrate pH 6.0 (PMC6993528).
Section 2
What Is the Expected CYP7B1 Staining Pattern?
CYP7B1 is an endoplasmic reticulum and microsome membrane protein with two annotated transmembrane segments (UniProt O75881 topology). In paraffin section IHC, expect predominantly cytoplasmic staining in cells that express it, including glandular, respiratory epithelial, hematopoietic and neuronal cells listed as high by HPA (HPA tissue IHC). Interpret that expectation cautiously: HPA also reports nuclear staining, but rates its tissue IHC evidence Uncertain because presumed off target binding was observed (HPA tissue IHC: Uncertain).
| Cytoplasmic staining in adrenal glandular cells or bronchial respiratory epithelial cells. | This fits the membrane associated location and HPA's high staining entries (UniProt O75881 subcellular location; HPA tissue IHC: High). Assess staining within the relevant cell population; HPA intensity is an observed category, not a guaranteed result for every section (HPA tissue IHC: Uncertain). |
| Strong nuclear staining with little cytoplasmic staining. | Treat a nuclear dominant result as suspect against the annotated endoplasmic reticulum location (UniProt O75881 subcellular location). HPA describes cytoplasmic and nuclear staining in most tissues, yet flags presumed off target binding; nuclear signal alone cannot establish CYP7B1 localisation (HPA tissue IHC: Uncertain). |
| Strong staining of soft tissue fibroblasts while the expected cells are weak. | HPA lists soft tissue fibroblasts as low, so this distribution warrants a specificity check (HPA tissue IHC: Low in soft tissue fibroblasts). Cross reactivity or endogenous chromogenic detection activity may produce apparent staining in an unexpected cell population (general IHC practice); morphology alone cannot identify which cause applies. |
| Broad colour over cells and tissue spaces, with no clear intracellular pattern. | This does not resolve the membrane associated, predominantly cytoplasmic pattern expected from UniProt (UniProt O75881 subcellular location). Diffuse colour can reflect nonspecific antibody binding, residual detection activity or inadequate washing (general IHC practice). Compare with a matched negative detection control before scoring cells as positive. |
| No staining in a section from a tissue HPA lists as high. | Adrenal gland, appendix, bone marrow, breast, bronchus, caudate, cerebellum and cerebral cortex contain listed high cell populations (HPA tissue IHC: High). First check that the relevant cells are present and the IHC run worked (general IHC practice). HPA's Uncertain reliability prevents treating a blank section as proof of absent CYP7B1 (HPA tissue IHC: Uncertain). |
💡Expected CYP7B1 appearanceCall a result provisionally positive when relevant HPA-listed cells show clear, predominantly cytoplasmic staining with limited background; HPA reports high staining in those listed populations but rates the IHC evidence Uncertain, and isolated nuclear or diffuse colour is suspect (UniProt O75881 subcellular location; HPA tissue IHC: High, Uncertain).
| Membrane topology | CYP7B1 has transmembrane segments at residues 17–37 and 289–309 and is annotated at endoplasmic reticulum and microsome membranes (UniProt O75881 topology). Use this location to assess compartment plausibility; the record does not define how finely a chromogenic section should resolve those membranes. |
| Tissue and cell selection | HPA reports high staining in specified glandular, respiratory epithelial, hematopoietic and neuronal populations, while soft tissue fibroblasts are low (HPA tissue IHC). Select and score the named cells within a section; a tissue label alone does not make every constituent cell an expected positive. |
| Confidence in the tissue pattern | HPA calls the tissue IHC evidence Uncertain, notes presumed off target binding was disregarded and says external verification is pending (HPA tissue IHC: Uncertain). Its high staining entries are comparison points, not independently validated positive controls. The listed antibody HPA017761 is also rated Uncertain for IHC (HPA antibody validation). |
| IF/ICC: what pattern is supported? | HPA gives a membrane summary but no main location, no ICC/IF images and no ICC validation status for HPA017761 (HPA subcellular; HPA antibody validation). UniProt supports an endoplasmic reticulum membrane expectation (UniProt O75881 subcellular location). These records cannot establish a validated IF/ICC pattern or settings. |
| Situation | Likely cause | Next action |
|---|---|---|
| The whole section is faint or blank, including an HPA-listed high cell population. | The relevant cells may be absent from the sampled area, or the staining run may have failed (general IHC practice). HPA's high category is uncertain evidence, not a guaranteed positive control (HPA tissue IHC: Uncertain). | Confirm cell identity and preservation on the section, then check the run's established control and detection steps (general IHC practice). Record a blank HPA-listed tissue as inconclusive until the assay works. |
| Signal is mainly nuclear. | Nuclear dominant colour conflicts with the annotated endoplasmic reticulum membrane location (UniProt O75881 subcellular location). HPA does report nuclear expression, but warns of presumed off target binding (HPA tissue IHC: Uncertain). | Compare nuclear and cytoplasmic signal in the same named cell population and inspect a matched negative detection control (general IHC practice). Avoid scoring nuclear staining alone as confirmed CYP7B1. |
| Fibroblasts stain as strongly as, or more strongly than, the intended cells. | Soft tissue fibroblasts are listed as low by HPA (HPA tissue IHC: Low in soft tissue fibroblasts). Nonspecific binding or endogenous detection activity can imitate cellular staining (general IHC practice). | Verify the stained cells by morphology, compare the matched negative detection control and review blocking and detection controls (general IHC practice). Treat the unexpected distribution as unresolved specificity. |
| A uniform chromogenic haze obscures cell boundaries. | Nonspecific binding, incomplete washing or endogenous detection activity can obscure a cellular pattern (general IHC practice). HPA's uncertain tissue reliability makes weak colour especially hard to interpret (HPA tissue IHC: Uncertain). | Inspect the negative detection control and review washing, blocking and chromogen development for the run (general IHC practice). Score only identifiable cells with signal separable from background. |
| Different tissues show different apparent staining strengths. | HPA's listed high populations span several tissues, and UniProt describes CYP7B1 as widely expressed (HPA tissue IHC: High; UniProt O75881 tissue specificity). Neither source supplies a quantitative intensity scale across preparations. | Compare the named cell populations, compartment and background within each section before comparing intensity across sections (general IHC practice). Do not infer expression differences from colour strength alone. |
| A suspected positive appears only with one antibody or one detection run. | HPA rates both its tissue IHC evidence and antibody HPA017761 as Uncertain; presumed off target binding was observed in the tissue assessment (HPA tissue IHC: Uncertain; HPA antibody validation). Run dependent signal is also possible (general IHC practice). | Repeat with appropriate run controls and seek independent specificity evidence before assigning CYP7B1 positivity (general IHC practice). Report the observed compartment and cell type without presenting the disputed signal as validated. |
Experiment evidence
IHC & IF images showing CYP7B1 staining
A03545-3 · IHCIHC analysis of CYP7B1 using anti-CYP7B1 antibody (A03545-3). CYP7B1 was detected in a paraffin-embedded section of human breast cancer tiss
HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000172817-CYP7B1/tissue/cerebral+cortex#imid_4966748
HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000172817-CYP7B1/tissue/colon#imid_4966666
HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000172817-CYP7B1/tissue/liver#imid_4966683
HPA · KidneyHuman Protein Atlas IHC, Kidney. Source: https://www.proteinatlas.org/ENSG00000172817-CYP7B1/tissue/kidney#imid_4966676
← swipe / scroll for more images →
Real IHC / IF data behind the staining-pattern expectations in this section.
Sample controls for CYP7B1 IHC & IF
🧪Run breast tissue first and assess staining in glandular cells, which HPA scores High (HPA: breast, glandular cells, High); the selected IHC caption also shows a paraffin section of human breast cancer (selected-SKU IHC caption: A03545-3). HPA detects CYP7B1 in all 45 scored tissues, so there is no HPA-supported negative tissue; use no-primary and isotype controls, and assess non-glandular cells on the positive slide for background staining without treating them as proven CYP7B1-negative cells (HPA: no negative rows; standard IHC practice).
Positive control tissue: Adrenal gland (Glandular cells, HPA High)
Negative control tissue: None in HPA: CYP7B1 is detected in all 45 scored tissues. Use a no-primary (secondary-only) and an isotype control instead.
ICC-IF cell lines (HPA subcellular resource): HPA carries no ICC-IF cell line for CYP7B1; derive a cell-line control from the positive tissue's cell type (Glandular cells) and confirm it by RNA or western blot first.
Technical controls: Run a no-primary, secondary-only slide and a rabbit isotype control matched to the primary antibody’s clonality where known; use CYP7B1-knockout tissue or a validated peptide-block condition as a biological specificity control (selected-SKU IHC caption: rabbit primary; standard IHC practice). Quench endogenous peroxidase and check background in the breast section before interpreting HRP–DAB staining (selected-SKU IHC caption: breast tissue, HRP–DAB detection; standard IHC practice).
⚠️Feasibility: No target-specific fixation window or fixation effect is reported, and the selected-SKU paraffin-section caption does not state its fixative (selected-SKU IHC caption: fixative unreported). That caption reports heat-mediated retrieval in EDTA at pH 8.0, but it does not establish whether retrieval is required under other conditions (selected-SKU IHC caption: EDTA pH 8.0). The supplied evidence demonstrates paraffin-section IHC but does not establish that frozen sections or IF are easier; breast-section interpretation should account for endogenous peroxidase background with HRP–DAB detection (selected-SKU IHC caption: paraffin breast section, HRP–DAB; standard IHC practice).
HPA tissue IHC evidence for CYP7B1
Comprehensive Human Protein Atlas IHC scoring per tissue (reliability: Uncertain — Presumed off target binding observed and disregarded. Pending external verification.). Rows are taken directly from the HPA tissue chart — click any row's HPA link to view the source.
Positive expression · recommended positive controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| Adrenal gland | Glandular cells | High | Protein (IHC) | HPA → |
| Appendix | Glandular cells | High | Protein (IHC) | HPA → |
| Bone marrow | Hematopoietic cells | High | Protein (IHC) | HPA → |
| Breast | Glandular cells | High | Protein (IHC) | HPA → |
| Bronchus | Respiratory epithelial cells | High | Protein (IHC) | HPA → |
Undetected expression · recommended negative controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| None in HPA: CYP7B1 is detected in all 45 scored tissues. Use a no-primary (secondary-only) and an isotype control instead. | ||||
Section 3
Advanced CYP7B1 IHC Tips
Troubleshoot CYP7B1 chromogenic IHC in paraffin sections by checking retrieval, staining compartment, controls, and scoring before interpreting biological differences.
Which retrieval conditions should I try first for weak CYP7B1 staining?
Start with heat mediated retrieval in EDTA at pH 8.0 for paraffin sections (datasheet A03545-3). The selected tissue IHC image used that buffer before an overnight incubation at 4°C with 2 μg/ml primary antibody (datasheet A03545-3). If staining remains weak, compare retrieval time and heating consistency across matched sections while holding antibody concentration and detection conditions fixed (standard IHC practice). Include a section stained under the documented conditions in the same run, and assess tissue damage as well as signal; a stronger DAB reaction alone does not establish specific CYP7B1 staining (standard IHC practice).
How should I troubleshoot variable CYP7B1 staining after tissue fixation?
Target specific fixation sensitivity is unknown: the selected CYP7B1 image describes a paraffin section but does not state its fixative (datasheet A03545-3). Record each specimen’s fixative, fixation duration, tissue thickness, and time before fixation, then compare sections processed and stained in the same run (standard IHC practice). If intensity varies, keep retrieval at EDTA pH 8.0 and primary antibody at 2 μg/ml while checking processing records (datasheet A03545-3). Do not assign a fixation effect from the reported tissue staining pattern or membrane topology; neither measures this antibody’s response to fixation (HPA: tissue IHC; UniProt O75881 topology).
Where should convincing CYP7B1 staining appear in a paraffin section?
Expect a predominantly intracellular, cytoplasmic pattern compatible with endoplasmic reticulum and microsome membranes (UniProt O75881 subcellular location). CYP7B1 has predicted transmembrane segments at residues 17–37 and 289–309, but the antibody epitope is not supplied, so topology cannot identify which side it detects (UniProt O75881 topology; datasheet A03545-3). HPA describes cytoplasmic and nuclear staining in many tissues, yet rates its tissue IHC reliability uncertain because presumed off target binding was observed (HPA: tissue IHC). Treat isolated nuclear or surface outlining signals cautiously, and compare cell morphology, negative controls, and reproducibility before calling them CYP7B1 (standard IHC practice; UniProt O75881 subcellular location).
Can an unreported epitope explain inconsistent CYP7B1 IHC staining?
Yes: epitope accessibility can change during paraffin processing and heat retrieval, while this antibody’s recognized sequence is not supplied here (standard IHC practice; datasheet A03545-3). UniProt lists a 506 residue chain, 2 transmembrane segments, no annotated isoforms, and no annotated glycosylation or modified residues (UniProt O75881). Those annotations do not establish where the antibody binds or prove that every staining difference reflects an epitope change (UniProt O75881; standard IHC practice). Keep EDTA at pH 8.0 as the reference condition, compare matched sections, and seek independent antibody or biological controls before attributing lost signal to epitope masking (datasheet A03545-3; standard IHC practice).
How can IF help assess an ambiguous CYP7B1 IHC pattern?
Use IF as a separate confirmation experiment alongside the chromogenic paraffin IHC result, with a marker for the expected cell population in the tissue being examined (standard IF practice; UniProt O75881 tissue specificity). Choose fluorophores after checking that tissue’s autofluorescence and include single stain controls when multiplexing (standard IF practice). CYP7B1 resides at endoplasmic reticulum and microsome membranes, with transmembrane segments at 17–37 and 289–309; select permeabilisation only after establishing which side contains the antibody epitope (UniProt O75881 topology; standard IF practice). The supplied records give no CYP7B1 ICC/IF images or epitope position, so evaluate IF localisation independently of the IHC caption (HPA: subcellular; datasheet A03545-3).
What should I check when CYP7B1 DAB staining is diffuse?
First inspect a no primary control for endogenous peroxidase activity and detection reagent background, and apply a peroxidase block as part of the chromogenic workflow (standard IHC practice). The selected image used 10% goat serum blocking, 2 μg/ml primary antibody overnight at 4°C, and peroxidase based DAB detection (datasheet A03545-3). Compare matched sections after adjusting primary concentration or wash stringency one variable at a time, while retaining the documented EDTA pH 8.0 retrieval reference (datasheet A03545-3; standard IHC practice). Because HPA reports presumed off target binding and uncertain tissue IHC reliability, diffuse signal needs independent specificity controls before interpretation (HPA: tissue IHC).
How should I score CYP7B1 staining across paraffin sections? ⚠ ANSWER MARKED FOR VERIFICATION
Define the cell population and acceptable intracellular staining pattern before scoring, then use the same regions and thresholds across specimens (standard IHC practice; UniProt O75881 subcellular location). For cell based analysis, report the percentage of positive cells and an H score from 0–300 using intensity categories applied consistently (standard IHC practice). If counting discrete positive cells, report density per mm² of evaluable tissue and normalise to the sampled area or relevant cell population (standard IHC practice). Exclude necrosis and section edges, document background subtraction, and interpret numerical differences cautiously because HPA rates CYP7B1 tissue IHC reliability uncertain (standard IHC practice; HPA: tissue IHC).
When is a CYP7B1 positive IHC result biologically credible?
A credible result shows reproducible intracellular staining in morphologically identifiable cells, consistent with CYP7B1’s endoplasmic reticulum and microsome membrane location (UniProt O75881 subcellular location; standard IHC practice). Compare the stained population with tissue context: UniProt reports broad expression, while HPA lists high staining in several cell populations but labels its tissue IHC reliability uncertain (UniProt O75881 tissue specificity; HPA: tissue IHC). Be cautious with isolated nuclear staining, unexpected cell types, section edge enhancement, or necrotic areas (HPA: tissue IHC; standard IHC practice). Check no primary and endogenous peroxidase controls, and seek independent specificity evidence before interpreting a DAB signal as CYP7B1 dependent biology (standard IHC practice; HPA: tissue IHC).
Boster reagents
Best CYP7B1 / Cytochrome P450 7B1 IHC Antibodies
A03545-3 has an IHC image from a human breast cancer paraffin section (IHC caption); the catalog lists human, mouse and rat reactivity (catalog: reactivity). No IF image is supplied (catalog: image data).
Real IHC data
Anti-CYP7B1 Antibody ®
Cat # A03545-3
A03545-3 will render with its human breast cancer paraffin-section IHC image (IHC caption). Its catalog lists IHC among the applications and human, mouse and rat reactivity, but supplies no IF image (catalog: applications, reactivity and image data).
Which to pick: Choose A03545-3 for paraffin-section tissue IHC: its caption documents heat retrieval in EDTA at pH 8.0 and 2 μg/ml primary antibody overnight at 4°C; the fixative is unreported (IHC caption). For IF/ICC, the catalog provides neither an IF application nor an IF image for A03545-3 (catalog: applications and image data). Although A03545-3 lists mouse and rat reactivity, its pictured IHC result and stated IHC dilution cover human tissue only (catalog: reactivity and dilution; IHC caption).
Each figure is that product's own IHC / IF validation image from its datasheet.
References
- UniProt Consortium. UniProt entry O75881 (CP7B1_HUMAN, Cytochrome P450 7B1).
- Human Protein Atlas. CYP7B1 tissue IHC expression (reliability: Uncertain).
- Human Protein Atlas. CYP7B1 subcellular location (ICC-IF): Membrane.
- Human Protein Atlas. CYP7B1 antibody validation summary (1 antibodies).
- Dehydroepiandrosterone and Its CYP7B1 Metabolite 7α-Hydroxydehydroepiandrosterone Regulates 11β-Hydroxysteroid Dehydrogenase 1 Directions in Rat Leydig Cells. Frontiers in endocrinology 2019 — PMC6993528.
- A Blunted GPR183/Oxysterol Axis During Dysglycemia Results in Delayed Recruitment of Macrophages to the Lung During Mycobacterium tuberculosis Infection. The Journal of infectious diseases 2022 — PMC9200159.
- CYP7B1 as a Biomarker for Prostate Cancer Risk and Progression: Metabolic and Oncogenic Signatures (Diagnostic Immunohistochemistry Analysis by Tissue Microarray in Prostate Cancer Patients-Diamond Study). International journal of molecular sciences 2024 — PMC11083792.
- 27-Hydroxycholesterol represses G9a expression via oestrogen receptor alpha in breast cancer. Journal of cellular and molecular medicine 2023 — PMC10494299.
- PubMed PMID:9802883 — UniProt-cited evidence.
- PubMed PMID:10588945 — UniProt-cited evidence.
- PubMed PMID:15489334 — UniProt-cited evidence.
Immunohistochemistry protocol sheet for CYP7B1 (Cytochrome P450 7B1). Expected localisation Cytoplasmic/nuclear staining (HPA tissue IHC); ER membrane (UniProt). Antigen retrieval: EDTA pH 8.0 HIER, heat-mediated (datasheet A03545-3). Validated IHC antibody: A03545-3 at 2-5 μg/ml (the product's own IHC validation image is shown). Controls (HPA): positive Adrenal gland (Glandular cells); negative None in HPA (detected in all 45 tissues); use no-primary + isotype controls. IHC-P workflow on paraffin sections; fixative unspecified. Protocol steps: 1. Sample: Paraffin-embedded human breast cancer tissue; fixative not specified (datasheet A03545-3); 2. Fixation: Image fixative and duration unreported (datasheet A03545-3); 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: EDTA pH 8.0 (datasheet A03545-3); 20 min, 95–100 °C (standard); 6. Peroxidase block: 3% H2O2, 10 min , room temperature (standard); 7. Blocking: 10% goat serum (datasheet A03545-3); 8. Primary antibody: Rabbit anti-CYP7B1, 2-5 μg/ml (datasheet A03545-3); 9. Primary incubation: Overnight at 4 °C (datasheet A03545-3). Watch for: Fixation: Keep paraffin-section fixation consistent (standard IHC practice; not target-specific); Caveat: Presumed off-target binding; reliability uncertain (HPA tissue IHC); Isoform / epitope: No annotated isoforms; one 1–506 chain (UniProt); Regulation: Regulation not specified (UniProt). Evidence: UniProt O75881, HPA tissue IHC · Uncertain, HPA ICC-IF · Membrane, Datasheet A03545-3, PMC6993528. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/CYP7B1/immunohistochemistry-ihc-guide.