CYP1B1 / Cytochrome P450 1B1 · IHC design guide

Design Immunohistochemistry for CYP1B1

Plan chromogenic CYP1B1 IHC-P around cytoplasmic staining in tissue sections (HPA tissue IHC). Use kidney tubules as a high-staining reference and colon glandular cells as a low-staining comparison (HPA tissue IHC), then optimize the catalog antibody within 0.5–1 μg/ml (datasheet PB9546).

Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use

Immunohistochemistry protocol sheet for CYP1B1 (IHC for CYP1B1): expected localisation Mainly cytoplasmic in tissue sections (HPA tissue IHC), antibody PB9546, validated IHC image, and IHC protocol steps — Printable **CYP1B1** IHC protocol sheet — expected localisation Mainly cytoplasmic in tissue sections (HPA tissue IHC), antibody PB9546, controls and protocol steps. Open the full CYP1B1 IHC guide →

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CYP1B1 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	Mainly cytoplasmic in tissue sections (HPA tissue IHC)
	Staining pattern	Cytoplasmic staining in kidney tubules and neurons (HPA tissue IHC)
	Antigen retrieval	Citrate pH 6 HIER, heat-mediated (datasheet PB9546)
	Positive control ⓘ	Cerebral cortex+4 more · see all
	Negative control ⓘ	Colon+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); its effect on staining is uncertain.
	Caveat	Antibody staining has low consistency with RNA expression (HPA tissue IHC)
	Regulation	No staining-intensity regulator documented (UniProt)
	Isoform / epitope	No annotated isoforms or processing; epitope map unresolved (UniProt)

Section 1

Recommended CYP1B1 IHC & IF Protocols

The catalog antibody’s IHC-P protocol uses citrate pH 6 retrieval (datasheet: PB9546). Three published chromogenic IHC protocols provide tissue-specific examples (PMC10167314; PMC12567446; PMC10879803).

Recommended immunohistochemistry (IHC-P) protocol parameters
Sample	Paraffin-embedded Mouse Kidney tissue; fixative not specified (datasheet PB9546)
Fixation	Image fixative and duration unreported (datasheet PB9546); verify before use.
Sectioning	4–5 µm sections on charged slides (standard)
Deparaffinisation	Xylene, graded ethanol series to water (standard)
Antigen retrieval	Heat retrieval: Citrate pH 6, 20 min (datasheet PB9546)
Peroxidase block	3% H2O2, 10 min, room temperature (standard)
Blocking	10% goat serum (datasheet PB9546)
Primary antibody	Rabbit anti-CYP1B1, 0.5-1μg/ml (datasheet PB9546)
Primary incubation	Overnight at 4 °C (datasheet PB9546)
Detection	Streptavidin-biotin complex (SABC), DAB chromogen (datasheet PB9546)
Counterstain	Hematoxylin, blue, dehydrate and mount (standard)
Expected result	CYP1B1-positive staining in neuronal cells of cerebral cortex (HPA tissue IHC: High). HPA tissue profile: Mainly cytoplasmic expression in most tissues. No signal in the no-primary control.

💡Decision noteStart with heat-mediated citrate pH 6 retrieval (datasheet: PB9546); adjust heating conditions for the specimen using the published protocols as references (PMC10167314; PMC12567446; PMC10879803).

Section 2

What Is the Expected CYP1B1 Staining Pattern?

CYP1B1 should appear mainly cytoplasmic in tissue IHC, with strong staining reported in cerebral cortex neurons, kidney tubular cells, and skeletal muscle myocytes (HPA tissue IHC). It is primarily associated with the endoplasmic reticulum and has no annotated transmembrane segment (UniProt Q16678 localisation and topology). Interpret intensity cautiously: HPA rates the tissue profile Approved but reports low consistency between antibody staining and RNA expression data (HPA tissue IHC).

What am I looking at on my slide?
Cytoplasmic staining in kidney tubular cells, cerebral cortex neurons, or skeletal muscle myocytes.	This fits HPA's mainly cytoplasmic tissue pattern and its High staining calls for these cell types (HPA tissue IHC). Assess the specified cells, rather than treating every stained structure in the section as an expected positive.
Predominantly nuclear or sharply cell-surface staining, without a convincing cytoplasmic pattern.	That differs from the reported tissue IHC pattern (HPA tissue IHC) and from CYP1B1's primary endoplasmic reticulum location (UniProt Q16678 localisation). Treat it as a possible artefact and check controls before assigning it to CYP1B1.
Strong staining in colon glandular cells or splenic red-pulp cells.	Those specific cells are Not detected in the HPA tissue IHC profile (HPA tissue IHC). Unexpected signal may reflect cross-reactivity or endogenous detection activity; inspect the negative control and staining distribution before interpreting it as expression.
Diffuse colour across cells, stroma, and tissue edges.	A widespread pattern is difficult to reconcile with the mainly cytoplasmic cell-associated profile (HPA tissue IHC). In a chromogenic IHC workflow, inspect blocking, washing, and the no-primary control for nonspecific signal or endogenous detection activity (general IHC practice).
No signal in kidney tubular cells, cerebral cortex neurons, or skeletal muscle myocytes.	These are High-staining cell types in the HPA tissue IHC profile (HPA tissue IHC). First check whether the appropriate cells are present and the assay controls worked; an absent signal alone cannot establish that the specimen lacks CYP1B1.

💡Expected CYP1B1 appearanceA credible positive is mainly cytoplasmic staining in the specified High-staining neurons, tubular cells, or myocytes (HPA tissue IHC); isolated nuclear, sharply surface-bound, or uniform cell-free colour warrants control review (general IHC interpretation).

How each factor affects the staining
Tissue and cell selection	HPA reports High staining in cerebral cortex neurons, kidney tubular cells, and skeletal muscle myocytes, but Not detected in colon glandular cells (HPA tissue IHC). Score the named cell population, since a tissue-level label can conceal different cellular patterns.
Evidence strength	HPA calls the tissue profile Approved while reporting low consistency between antibody staining and RNA expression data (HPA tissue IHC). Its IHC antibody entries HPA026863 and CAB011705 are Approved, without an Enhanced designation in the supplied record (HPA antibodies).
Compartment and assay context	UniProt places CYP1B1 primarily at the endoplasmic reticulum and also notes mitochondrial targeting after TCDD treatment (UniProt Q16678 localisation). HPA tissue IHC is mainly cytoplasmic, whereas its ICC-IF mitochondrial assignment is uncertain (HPA tissue IHC; HPA subcellular).
Topology and molecular annotations	UniProt lists no transmembrane segment, signal peptide, propeptide, annotated isoforms, glycosylation sites, or modified residues (UniProt Q16678 topology and annotations). Those entries do not establish an optimal retrieval condition or a target-specific fixation effect.
Retrieval and detection settings	Target-specific fixation effects are not established by the supplied assay evidence. Verify with a matched IHC source before attributing a result to fixation.

Why is my staining missing, weak or wrong?
Situation	Likely cause	Next action
Known High-staining cells show no chromogenic signal.	A missed cell population or failed staining run is possible; the expected High calls apply to specified cells (HPA tissue IHC).	Confirm those cells are present, then review the run control, antibody dilution, retrieval, and detection steps against the catalog IHC-P instructions (general IHC practice).
Colon glandular cells stain strongly.	HPA reports these cells as Not detected (HPA tissue IHC); cross-reactivity or endogenous detection activity is possible.	Compare a no-primary control and the cell-level pattern, then repeat with an independently validated IHC antibody if the finding matters (general IHC practice).
Signal is mainly nuclear or sharply outlines the cell surface.	The pattern conflicts with mainly cytoplasmic tissue staining (HPA tissue IHC) and the primary endoplasmic reticulum location (UniProt Q16678 localisation).	Check counterstain and chromogen localisation, inspect controls, and verify the pattern with an independent IHC antibody before scoring it (general IHC practice).
Colour covers stroma or tissue edges as well as cells.	Diffuse deposition obscures the cell-associated pattern described by HPA (HPA tissue IHC); nonspecific detection is possible (general IHC practice).	Review the no-primary control, blocking, washes, and chromogen development; score only interpretable cellular staining (general IHC practice).
ICC-IF appears mitochondrial while paraffin IHC appears cytoplasmic.	HPA's ICC-IF mitochondrial location is uncertain (HPA subcellular); its tissue IHC profile is mainly cytoplasmic (HPA tissue IHC). UniProt also describes conditional mitochondrial targeting after TCDD treatment (UniProt Q16678 localisation).	Keep the assay interpretations separate. Use the dedicated IF/ICC guide for IF controls and localisation; judge this IHC result against tissue IHC controls and cell-level pattern.
A weakly stained section is hard to classify.	HPA reports low consistency between antibody staining and RNA expression data despite an Approved tissue profile (HPA tissue IHC). Weak colour alone may therefore be inconclusive.	Compare matched positive and no-primary controls, confirm the intended cells, and record intensity with its distribution before calling the specimen positive or negative (general IHC practice).

Source: UniProt Q16678 Human Protein Atlas tissue Human Protein Atlas subcellular

Experiment evidence

IHC & IF images showing CYP1B1 staining

IHC analysis of CYP1B1 using anti-CYP1B1 antibody (PB9546). CYP1B1 was detected in paraffin-embedded section of Mouse Kidney Tissue. Heat mediated antigen retrieval was performed in citrate buffer (pH6, epitope retrieval solution) for 20 mins. The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1μg/ml rabbit anti-CYP1B1 Antibody (PB9546) overnight at 4°C. Biotinylated goat anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC)(Catalog # SA1022) with DAB as the chromogen.

PB9546 · IHCIHC analysis of CYP1B1 using anti-CYP1B1 antibody (PB9546). CYP1B1 was detected in paraffin-embedded section of Mouse Kidney Tissue. Heat me

IHC analysis of CYP1B1 using anti-CYP1B1 antibody (PB9546). CYP1B1 was detected in paraffin-embedded section of Rat Kidney Tissue. Heat mediated antigen retrieval was performed in citrate buffer (pH6, epitope retrieval solution) for 20 mins. The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1μg/ml rabbit anti-CYP1B1 Antibody (PB9546) overnight at 4°C. Biotinylated goat anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC)(Catalog # SA1022) with DAB as the chromogen.

PB9546 · IHCIHC analysis of CYP1B1 using anti-CYP1B1 antibody (PB9546). CYP1B1 was detected in paraffin-embedded section of Rat Kidney Tissue. Heat medi

HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000138061-CYP1B1/tissue/cerebral+cortex#imid_7375440

HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000138061-CYP1B1/tissue/colon#imid_7374078

HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000138061-CYP1B1/tissue/liver#imid_7374079

HPA · KidneyHuman Protein Atlas IHC, Kidney. Source: https://www.proteinatlas.org/ENSG00000138061-CYP1B1/tissue/kidney#imid_7374080

HPA · ICC-IF A-549Human Protein Atlas immunofluorescence, A-549, antibody HPA026863; green = CYP1B1, blue = nucleus, red = microtubules.

← swipe / scroll for more images →

Real IHC / IF data behind the staining-pattern expectations in this section.

Sample controls for CYP1B1 IHC & IF

🧪Run kidney first and assess staining in tubular cells (HPA: High in kidney tubule cells). Use colon glandular cells as the negative tissue (HPA: Not detected in colon glandular cells); unstained cells within the kidney section, where present, should show counterstain without specific DAB deposit (standard IHC practice).

Positive control tissue: Cerebral cortex (Neuronal cells, HPA High)

Negative control tissue: Colon (HPA Not detected)

ICC-IF cell lines (HPA subcellular resource): HPA ICC-IF images show CYP1B1 in A-549, SiHa, U2OS, with annotated localisation: Mitochondria (uncertain) (HPA subcellular).

Technical controls: Include a no-primary (secondary-only) slide, a rabbit isotype control matched to the primary antibody’s immunoglobulin class and clonality where applicable, and CYP1B1-knockout tissue if available (standard IHC practice; caption: rabbit anti-CYP1B1). In kidney, block endogenous peroxidase and check for endogenous biotin because the reported detection uses a biotinylated secondary, streptavidin–biotin complex and DAB (standard IHC practice; PB9546 tissue-IHC caption).

⚠️Feasibility: The PB9546 paraffin-section caption reports heat retrieval in citrate buffer at pH 6 for 20 min in mouse kidney, but does not establish whether retrieval is required (PB9546 tissue-IHC caption). A target-specific fixation window or fixation effect is unreported, and the selected-SKU caption does not report a fixative (PB9546 tissue-IHC caption). The supplied evidence does not establish that frozen sections or IF are easier; renal endogenous peroxidase or biotin may complicate interpretation of the reported DAB method (standard IHC practice; PB9546 tissue-IHC caption).

HPA tissue IHC evidence for CYP1B1

Comprehensive Human Protein Atlas IHC scoring per tissue (reliability: Approved — Low 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
Cerebral cortex	Neuronal cells	High	Protein (IHC)	HPA →
Kidney	Cells in tubules	High	Protein (IHC)	HPA →
Skeletal muscle	Myocytes	High	Protein (IHC)	HPA →
Adipose tissue	Adipocytes	Medium	Protein (IHC)	HPA →
Adrenal gland	Glandular cells	Medium	Protein (IHC)	HPA →

Undetected expression · recommended negative controls

Tissue	Cell type	Level	Evidence	Source
Colon	Glandular cells	Not detected	Protein (IHC)	HPA →
Duodenum	Glandular cells	Not detected	Protein (IHC)	HPA →
Lymph node	Germinal center cells	Not detected	Protein (IHC)	HPA →
Oral mucosa	Squamous epithelial cells	Not detected	Protein (IHC)	HPA →
Spleen	Cells in red pulp	Not detected	Protein (IHC)	HPA →

Section 3

Advanced CYP1B1 IHC Tips

Troubleshoot CYP1B1 staining in paraffin sections by checking retrieval, controls, cellular localisation and scoring before interpreting chromogenic signal.

What retrieval should I start with if CYP1B1 staining is weak?

Start with heat-mediated retrieval in citrate buffer at pH 6 for 20 minutes (datasheet PB9546). The catalog antibody detected CYP1B1 in a paraffin-embedded mouse kidney section after this treatment, followed by 1 μg/ml primary antibody overnight at 4°C (caption PB9546). If signal remains weak, compare retrieval times on adjacent sections while holding antibody concentration and detection conditions constant (standard IHC practice). Check whether improved signal appears in kidney tubular cells, where HPA reports high staining, rather than judging retrieval by overall section darkness (HPA: High in kidney tubular cells).

Could fixation explain weak or uneven CYP1B1 staining?

Target-specific fixation sensitivity is unknown: the selected paraffin-section caption does not state a fixative, so it cannot establish how CYP1B1 responds to fixation (caption PB9546). Record the fixative and processing history for each section before comparing staining across samples (standard IHC practice). Compare sections processed under matched conditions and begin with citrate retrieval at pH 6 for 20 minutes, the documented starting condition for this antibody (datasheet PB9546). If uneven staining follows tissue edges or processing boundaries, examine section quality and include a matched control before attributing the pattern to CYP1B1 biology (standard IHC practice).

Which staining pattern fits CYP1B1 in paraffin sections?

Assess CYP1B1 primarily as cytoplasmic staining in tissue IHC, while allowing a granular or perinuclear pattern consistent with its predominant endoplasmic reticulum localisation (HPA: mainly cytoplasmic expression; UniProt Q16678 localisation). UniProt also lists microsomal and mitochondrial locations, with mitochondrial targeting reported after TCDD treatment (UniProt Q16678 localisation). A solely nuclear or extracellular DAB pattern therefore warrants a check of morphology, detection background and controls before it is scored as positive (UniProt Q16678 localisation; standard IHC practice). Compare the pattern within identified cells, such as kidney tubular cells, instead of treating diffuse brown colour across the section as localisation evidence (HPA: High in kidney tubular cells; standard IHC practice).

How should I troubleshoot uncertain epitope access or apparent isoform differences?

The supplied record annotates one 543-amino-acid chain and no isoforms, glycosylation sites or modified residues; it does not identify this antibody’s epitope (UniProt Q16678 record; caption PB9546). Do not assign distinct tissue staining patterns to specific CYP1B1 isoforms or post-translational modifications on this evidence (UniProt Q16678 record). The record reports membrane-associated locations but no transmembrane segment, so the epitope’s accessibility in a paraffin section remains an empirical question (UniProt Q16678 topology and localisation). Compare retrieval and primary-antibody omission controls on adjacent sections, then judge any gain in signal against cellular morphology and the expected cytoplasmic distribution (datasheet PB9546; HPA: mainly cytoplasmic expression; standard IHC practice).

How can IF help resolve ambiguous chromogenic CYP1B1 staining?

On a separate IF/ICC preparation, pair CYP1B1 with a marker that identifies the cell population under study; kidney tubular cells are one IHC-supported population for comparison (HPA: High in kidney tubular cells; standard IF practice). Choose fluorophores after inspecting unstained tissue autofluorescence and include single-channel controls when testing overlap (standard IF practice). Because the supplied record gives no antibody epitope or membrane-facing orientation, optimise permeabilisation empirically for access to the epitope rather than assuming its side of a membrane (UniProt Q16678 topology; standard IF practice). Treat mitochondrial overlap cautiously: HPA calls that IF localisation uncertain, while UniProt describes CYP1B1 as primarily endoplasmic reticulum-associated (HPA subcellular: mitochondria, uncertain; UniProt Q16678 localisation).

What should I check when DAB background obscures CYP1B1?

First inspect a section processed without primary antibody to identify signal from the detection system, and check whether brown staining follows tissue edges or damaged areas (standard IHC practice). Include a peroxidase-blocking step before chromogenic detection and verify that its omission control is clean; this is a general DAB workflow check, not CYP1B1-specific evidence (standard IHC practice). The selected tissue caption used 10% goat-serum block, 1 μg/ml primary antibody and DAB detection, providing a documented starting condition for PB9546 (caption PB9546). If background persists, adjust blocking, washing or antibody concentration one variable at a time and retain a morphology-matched positive control (standard IHC practice).

How should I quantify CYP1B1 staining across paraffin sections? ⚠ ANSWER MARKED FOR VERIFICATION

Define the cell compartment and region of interest before scoring, then record either an H-score or the percentage of positive cells within that defined population (standard IHC practice). For H-scores, grade staining intensity from 0–3 and sum each grade multiplied by its percentage of cells, yielding 0–300 (standard IHC practice). If counting positive cells per mm², normalise to the analysed tissue area and report the cell type and exclusion criteria alongside the density (standard IHC practice). Keep retrieval, imaging and threshold settings matched across slides, and interpret kidney tubular-cell scores separately from other renal compartments (datasheet PB9546; HPA: High in kidney tubular cells; standard IHC practice).

How do I distinguish convincing CYP1B1 staining from artefact?

A convincing positive has cellular boundaries and a mainly cytoplasmic pattern in an identified cell population; HPA reports high staining in kidney tubular cells (HPA: mainly cytoplasmic expression; HPA: High in kidney tubular cells). Purely nuclear or extracellular colour conflicts with the supplied localisation evidence and should prompt review of morphology and controls (UniProt Q16678 localisation; standard IHC practice). Exclude edge staining, necrotic areas and signal that persists without primary antibody, and check peroxidase-related DAB background before scoring (standard IHC practice). Interpret differences between tissues cautiously because HPA rates its tissue IHC evidence Approved but reports low consistency between antibody staining and RNA expression (HPA: reliability description).

Boster reagents

Best CYP1B1 / Cytochrome P450 1B1 IHC Antibodies

CYP1B1 antibodies PB9546 and M00515 list IHC and human, mouse, and rat reactivity (catalog applications and reactivity); PB9546 has paraffin section IHC figures, while neither lists IF/ICC (catalog applications and image captions).

Real IHC data

Anti-CYP1B1 Antibody ®

Cat # PB9546

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PB9546 has IHC figures from paraffin sections of mouse kidney, rat kidney, and human liver cancer tissue (PB9546 image captions). M00515 lists IHC and human, mouse, and rat reactivity, but provides no IHC figure in this payload (M00515 catalog applications, reactivity, and image captions).

Which to pick: Choose PB9546 for tissue IHC when a matching paraffin section example matters; its captions show those three tissues, and its fixative is unreported (PB9546 image captions). For cross species IHC, both SKUs list human, mouse, and rat reactivity, but only PB9546 has figures across those species; M00515 is monoclonal and PB9546 is listed without a clone (catalog reactivity, clone fields, and PB9546 image captions). Neither SKU lists IF/ICC validation, so there is no supported IF/ICC pick here (catalog applications).

Each figure is that product's own IHC / IF validation image from its datasheet.

References

UniProt Consortium. UniProt entry Q16678 (CP1B1_HUMAN, Cytochrome P450 1B1).
Human Protein Atlas. CYP1B1 tissue IHC expression (reliability: Approved).
Human Protein Atlas. CYP1B1 subcellular location (ICC-IF): Localized to the mitochondria..
Human Protein Atlas. CYP1B1 antibody validation summary (2 antibodies).
Hepatic stellate cell stearoyl co-A desaturase activates leukotriene B4 receptor 2 - β-catenin cascade to promote liver tumorigenesis. Nature communications 2023 — PMC10167314.
Expression and Clinical Significance of Cytochrome 1B1 in Bone Sarcomas. Pharmaceuticals (Basel, Switzerland) 2025 — PMC12567446.
Evaluation of CYP1B1, oxidative stress and phase II detoxification enzyme status in oral cancer progression model. Journal of oral biology and craniofacial research 2024 — PMC10879803.
Role of mammary epithelial and stromal P450 enzymes in the clearance and metabolic activation of 7,12-dimethylbenz(a)anthracene in mice. Toxicology letters 2012 — PMC3668431.
PubMed PMID:8175734 — UniProt-cited evidence.
PubMed PMID:8910454 — UniProt-cited evidence.
PubMed PMID:15489334 — UniProt-cited evidence.