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CYP7B1 / Cytochrome P450 7B1 · IHC design guide
Design Immunohistochemistry for CYP7B1
Plan CYP7B1 chromogenic IHC on paraffin sections using the catalog antibody’s 2–5 μg/ml range (datasheet A03545-3). Assess cytoplasmic and nuclear staining with caution because the tissue IHC evidence flags possible off-target binding (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 | Glandular, neuronal and hematopoietic cells: cytoplasmic/nuclear (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 fixation consistent across paraffin sections (standard IHC practice; not target-specific) | |
| Caveat | Possible off-target binding; staining reliability is uncertain (HPA tissue IHC) | |
| Regulation | No expression regulator specified (UniProt) | |
| Isoform / epitope | No annotated isoforms or processing; epitope side unspecified (UniProt) |
Section 1
Recommended CYP7B1 IHC & IF Protocols
The catalog antibody’s IHC-P protocol (datasheet A03545-3) is accompanied by three published CYP7B1 staining protocols (PMC6993528; PMC9200159; PMC10494299).
| 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 EDTA pH 8.0 heat retrieval for the catalog antibody (datasheet A03545-3); the published protocols use different retrieval methods (PMC6993528; PMC9200159; PMC10494299).
Section 2
What Is the Expected CYP7B1 Staining Pattern?
CYP7B1 is an endoplasmic reticulum and microsome membrane protein with transmembrane segments at residues 17–37 and 289–309 (UniProt O75881 topology). In paraffin-section IHC, assess cytoplasmic staining in the cell populations HPA reports as positive, including adrenal glandular cells, bone marrow hematopoietic cells and cerebral cortex neurons (HPA tissue IHC). HPA also reports nuclear staining in many tissues, but rates its tissue IHC evidence Uncertain because presumed off-target binding was observed (HPA tissue IHC).
| Cytoplasmic staining appears in adrenal glandular cells, bone marrow hematopoietic cells or cerebral cortex neurons. | These are reported high-staining cell populations (HPA: High in each). A cell-associated cytoplasmic pattern is compatible with CYP7B1 at the endoplasmic reticulum and microsome membranes (UniProt O75881 subcellular location). HPA rates the tissue IHC evidence Uncertain, so a matching pattern supports interpretation but does not establish antibody specificity (HPA tissue IHC). |
| Signal is predominantly nuclear, with little or no cytoplasmic staining. | Interpret cautiously: UniProt places CYP7B1 at endoplasmic reticulum and microsome membranes (UniProt O75881 subcellular location), while HPA describes both cytoplasmic and nuclear staining in most tissues (HPA tissue IHC). Because HPA disregarded presumed off-target binding and assigns Uncertain reliability, nuclear staining alone is insufficient to identify specific CYP7B1 signal (HPA tissue IHC). |
| A different cell population stains more strongly than the reported positive cells. | Compare cell identities before assigning the signal to CYP7B1: HPA reports high staining in defined glandular, hematopoietic and neuronal populations, among others (HPA tissue IHC). Unexpected staining may reflect cross-reactivity or endogenous detection activity (general IHC practice). The HPA list is illustrative, so staining outside those examples is not, by itself, proof of an artefact (HPA tissue IHC). |
| Color is widespread across the section without clear cell boundaries or a compartment pattern. | Diffuse deposition is difficult to reconcile with the endoplasmic reticulum and microsome membrane location (UniProt O75881 subcellular location). Nonspecific antibody binding or detection background can produce diffuse chromogenic signal (general IHC practice). Judge the pattern against a matched negative detection control and the reported positive cells before scoring it as CYP7B1 (general IHC practice; HPA tissue IHC). |
| No signal appears in an expected high-staining population, such as adrenal glandular cells. | HPA reports High staining in adrenal glandular cells (HPA tissue IHC), so an absent signal warrants a check of section quality, retrieval, antibody incubation and chromogenic detection (general IHC practice). It does not alone establish that the tissue lacks CYP7B1; HPA's tissue IHC reliability is Uncertain (HPA tissue IHC). |
💡Expected CYP7B1 appearanceCall a result provisionally positive when cell-associated cytoplasmic staining is visible in an HPA high-staining population, such as adrenal glandular cells; interpret nuclear-only, unexpected-cell or diffuse signal cautiously because the protein is membrane-associated and HPA tissue IHC reliability is Uncertain (UniProt O75881 subcellular location; HPA tissue IHC).
| Membrane location and epitope access | CYP7B1 has two annotated transmembrane segments and localizes to endoplasmic reticulum and microsome membranes (UniProt O75881 topology and subcellular location). Epitope accessibility depends on the antibody's epitope and the staining conditions (general IHC practice); neither an epitope position nor a CYP7B1-specific antigen retrieval requirement is supplied here. |
| Strength of tissue-pattern evidence | HPA reports high staining across several cell populations but rates its tissue IHC evidence Uncertain after disregarding presumed off-target binding (HPA tissue IHC). Its listed antibody, HPA017761, has IHC status Uncertain (HPA antibodies). Use the cell pattern as a comparison, with controls, rather than as independent proof of specificity (general IHC practice). |
| IF/ICC Q&A | Q: Can the IHC pattern be carried over to IF/ICC? A: UniProt supports a membrane-associated expectation (UniProt O75881 subcellular location), and HPA's subcellular summary says “Membrane” (HPA subcellular). HPA supplies no ICC-IF image cell lines, and its listed antibody has no ICC status (HPA subcellular; HPA antibodies), so an IF/ICC staining pattern cannot be validated from these records. |
| Situation | Likely cause | Next action |
|---|---|---|
| Adrenal glandular cells show no chromogenic signal. | The section or staining workflow may have failed (general IHC practice); HPA reports these cells as High staining (HPA tissue IHC). | Confirm tissue identity and section integrity, then check retrieval, antibody incubation and detection against appropriate controls (general IHC practice). Treat a persistently negative result cautiously because HPA tissue IHC reliability is Uncertain (HPA tissue IHC). |
| Nuclei stain strongly while cytoplasm is faint. | HPA reports nuclear as well as cytoplasmic staining, but also flags presumed off-target binding (HPA tissue IHC); UniProt places CYP7B1 at intracellular membranes (UniProt O75881 subcellular location). | Record the nuclear and cytoplasmic signals separately. Compare with a negative detection control and a reported positive cell population before assigning nuclear staining to CYP7B1 (general IHC practice; HPA tissue IHC). |
| Color appears in unexpected cells or across the whole section. | Cross-reactivity, nonspecific binding or endogenous detection activity can create misleading signal (general IHC practice). HPA reports specific high-staining cell populations but assigns Uncertain reliability (HPA tissue IHC). | Check cell identity and review matched negative controls; if using enzyme-based chromogenic detection, assess endogenous activity with an appropriate control (general IHC practice). Reassess any signal that does not track recognizable cells (general IHC practice). |
| Background obscures the difference between cells and surrounding tissue. | Nonspecific binding, excessive antibody signal or detection background can reduce contrast (general IHC practice); the supplied records do not identify a CYP7B1-specific cause. | Review blocking, washes, antibody concentration and detection exposure using the established IHC workflow and negative controls (general IHC practice). Score only signal that remains clearly cell-associated and interpretable (general IHC practice). |
| Bone marrow or cerebral cortex stains, but the expected cells are unclear. | HPA attributes High staining to bone marrow hematopoietic cells and cerebral cortex neuronal cells (HPA tissue IHC); mixed cell populations can complicate assignment on a section (general IHC practice). | Identify the staining cells from section morphology and counterstain before comparing intensity with HPA's cell-specific observations (general IHC practice; HPA tissue IHC). Avoid scoring the entire tissue as uniformly positive when the relevant cells cannot be resolved (general IHC practice). |
| An IF/ICC image seems to show a different compartment. | HPA gives a “Membrane” subcellular summary but provides no ICC-IF image cell lines or ICC status for its listed antibody (HPA subcellular; HPA antibodies). | Keep the IF/ICC observation separate from this paraffin IHC assessment. Compare it with UniProt's endoplasmic reticulum and microsome membrane location, and require assay-specific controls before assigning the fluorescent signal to CYP7B1 (UniProt O75881 subcellular location; general IF practice). |
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 first; glandular cells should stain strongly (HPA: breast glandular cells, High). HPA detects CYP7B1 in all 45 scored tissues, so there is no supported negative tissue; use no-primary and isotype controls for the negative comparison (HPA: no negative rows). On the positive slide, assess nearby nonglandular stromal cells as an internal background comparator: they should lack convincing ER-like cytoplasmic staining if they are negative, but their CYP7B1 status is not established by these rows (UniProt O75881: ER membrane; HPA: breast glandular cells, High).
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 control and a host- and immunoglobulin-class-matched rabbit IgG isotype control alongside the rabbit catalog antibody (selected-SKU caption: rabbit anti-CYP7B1). Use CYP7B1 knockout material, if available, as a biological specificity control; quench endogenous peroxidase before HRP/DAB detection and check blood or inflammatory cells for residual background (selected-SKU caption: HRP/DAB detection; standard IHC practice).
⚠️Feasibility: Paraffin-section IHC is supported by the selected-SKU breast cancer caption, which uses heat retrieval in EDTA at pH 8.0; the fixative is unreported, and no target-specific fixation window or fixation effect is supplied (selected-SKU caption). Retrieval is part of that demonstrated workflow, but its necessity has not been established by a comparison (selected-SKU caption: heat retrieval). The supplied evidence does not establish whether frozen sections or IF are easier; for breast sections, assess endogenous peroxidase background under the stated HRP/DAB detection conditions (selected-SKU caption: HRP/DAB detection; 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 staining in paraffin sections with attention to retrieval, membrane localisation, controls, and the limits of available tissue evidence.
Which retrieval conditions should I start with for CYP7B1 paraffin sections?
Start with heat-mediated retrieval in EDTA at pH 8.0 for this catalog antibody (datasheet A03545-3). The selected paraffin-section image used those conditions, followed by 10% goat serum blocking and 2 μg/ml primary antibody overnight at 4°C (datasheet A03545-3). If staining is weak, vary heating duration in a controlled series while keeping detection and section handling constant; excessive heating can damage morphology (standard IHC practice). Compare signal in viable tissue with a no-primary control and record both intensity and tissue preservation, because the image caption does not report retrieval time (datasheet A03545-3).
How should I handle fixation when CYP7B1 staining is weak or uneven?
Target-specific sensitivity to fixation is unknown: the selected CYP7B1 image specifies a paraffin-embedded section but does not state its fixative (datasheet A03545-3). Record the fixative, fixation interval, processing history, and section age for every specimen, then compare sections processed together before changing antibody concentration (standard IHC practice). In the same run, examine morphology, edge-to-center staining, and a no-primary control to distinguish preparation effects from detection background (standard IHC practice). If fixation histories differ, interpret intensity differences cautiously and optimize retrieval on matched material; neither the tissue staining profile nor the protein topology establishes a CYP7B1-specific fixation effect (HPA: tissue reliability Uncertain; UniProt O75881 topology).
Where should convincing CYP7B1 staining appear in tissue cells?
Expect CYP7B1-associated signal principally in a cytoplasmic membrane pattern consistent with endoplasmic reticulum and microsomes (UniProt O75881 subcellular location). Its annotated transmembrane segments span residues 17–37 and 289–309, supporting membrane association without specifying the antibody epitope or its accessible side (UniProt O75881 topology). Assess staining within clearly identified cells and compare the pattern across adjacent, well-preserved fields; a diffuse nuclear-only pattern needs independent confirmation (standard IHC practice; UniProt O75881 subcellular location). Although tissue images report cytoplasmic and nuclear expression, their reliability is marked Uncertain because presumed off-target binding was observed (HPA: tissue IHC reliability Uncertain).
Could isoforms or epitope accessibility explain discordant CYP7B1 staining?
The supplied record annotates a single 1–506 CYP7B1 chain and no isoforms, so an isoform-specific staining explanation has no support here (UniProt O75881 processing and isoforms). It lists 2 transmembrane segments, at residues 17–37 and 289–309, but provides no antibody epitope or membrane sidedness (UniProt O75881 topology). Ask for epitope mapping before assigning a staining loss to masking on one membrane face; compare retrieval conditions on matched sections instead (standard IHC practice). The record annotates no glycosylation sites or modified residues, so do not attribute variable staining to a documented CYP7B1 modification (UniProt O75881 PTMs).
How can IF help check the cell type and compartment stained by CYP7B1 IHC?
Use IF as a separate validation experiment, since the selected antibody evidence describes chromogenic paraffin-section IHC rather than an IF protocol (datasheet A03545-3). In breast sections, multiplex CYP7B1 with an epithelial marker to identify glandular cells, while treating the reported high glandular staining as provisional (HPA: breast glandular cells High; HPA: tissue reliability Uncertain). Choose a far-red fluorophore when green tissue autofluorescence obscures signal, and include single-color and no-primary controls (standard IF practice). If the epitope is mapped to a cytosolic or luminal membrane face, optimize permeabilisation accordingly; the supplied topology alone does not establish that face (UniProt O75881 topology).
What controls help separate CYP7B1 staining from chromogenic background?
The selected image used 10% goat serum blocking, a peroxidase-conjugated secondary antibody, and DAB development (datasheet A03545-3). Include a peroxidase block, no-primary section, and matched secondary-only assessment where appropriate to reveal endogenous enzyme activity or secondary binding (standard IHC practice). Keep DAB development time and counterstaining consistent across compared sections, and inspect folds, damaged edges, and necrotic regions before calling weak signal positive (standard IHC practice). Because the tissue profile was rated Uncertain after presumed off-target binding, a clean no-primary control alone cannot establish antibody specificity (HPA: tissue IHC reliability Uncertain).
How should I quantify CYP7B1 staining across paraffin sections? ⚠ ANSWER MARKED FOR VERIFICATION
Define the cell population and viable tissue region before scoring, then use the same segmentation and chromogen threshold for every slide (standard IHC practice). An H-score combines the percentage of cells at intensity grades 0–3 into a 0–300 summary; also report percentage positive cells or positive-cell density per mm² (standard IHC practice). Normalize cell counts to the number of evaluable target cells, and area-based counts to viable tissue area, while excluding folds and necrosis (standard IHC practice). Record compartment-specific scores because CYP7B1 is assigned to endoplasmic reticulum and microsome membranes, whereas reported nuclear tissue staining remains uncertain (UniProt O75881 subcellular location; HPA: tissue IHC reliability Uncertain).
When should I distrust an apparent CYP7B1-positive result?
Favor staining in intact cells with a cytoplasmic membrane distribution compatible with CYP7B1’s endoplasmic reticulum and microsome assignment (UniProt O75881 subcellular location). Question isolated nuclear signal, staining confined to section edges or necrosis, and DAB deposition reproduced by a no-primary control; assess endogenous peroxidase as a possible source (standard IHC practice; HPA: tissue IHC reliability Uncertain). Check cell identity against morphology and an appropriate marker rather than assuming every stained cell matches the reported tissue profile (standard IHC practice). The selected breast-cancer image demonstrates staining under its stated conditions, but the broader tissue atlas flags presumed off-target binding and pending external verification (datasheet A03545-3; HPA: tissue IHC reliability Uncertain).
Boster reagents
Best CYP7B1 / Cytochrome P450 7B1 IHC Antibodies
A03545-3 has real IHC data from a human breast cancer paraffin section (image caption). Human, mouse and rat reactivity is listed (catalog); IF/ICC validation is unreported (catalog).
Real IHC data
Anti-CYP7B1 Antibody ®
Cat # A03545-3
A03545-3 was demonstrated by IHC on a paraffin-embedded human breast cancer section (image caption). Its listed applications include IHC, and its listed reactivity includes human, mouse and rat (catalog).
Which to pick: Choose A03545-3 for paraffin-section IHC: its own figure shows EDTA retrieval, 2 μg/ml primary antibody and DAB detection; the fixative is unreported (image caption). IF/ICC validation is unreported, so there is no validated IF/ICC pick here (catalog). For mouse or rat studies, A03545-3 lists reactivity with both species, but its pictured IHC example is human; it is rabbit-hosted, and clonality is unreported (catalog; image 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 fixation consistent across paraffin sections (standard IHC practice; not target-specific); Caveat: Possible off-target binding; staining reliability is uncertain (HPA tissue IHC); Isoform / epitope: No annotated isoforms or processing; epitope side unspecified (UniProt); Regulation: No expression regulator specified (UniProt). Evidence: UniProt O75881, HPA tissue IHC · Uncertain, HPA ICC-IF · Membrane, Datasheet A03545-3, PMC6993528, PMC9200159. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/CYP7B1/immunohistochemistry-ihc-guide.