BRD7 / Bromodomain-containing protein 7 · IHC design guide

Design Immunohistochemistry for BRD7

Plan BRD7 IHC in paraffin sections around the broad nuclear staining reported in tissues (HPA tissue IHC). This guide covers consistent fixation, the catalog antibody’s 0.5–1 μg/ml IHC range (datasheet A01289-1), and interpretation of adipocytes with no detectable staining (HPA tissue IHC).

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

Immunohistochemistry protocol sheet for BRD7 (IHC for BRD7): expected localisation Nuclear staining in tissue (HPA tissue IHC); chromosome association (UniProt), antibody A01289-1, validated IHC image, and IHC protocol steps — Printable **BRD7** IHC protocol sheet — expected localisation Nuclear staining in tissue (HPA tissue IHC); chromosome association (UniProt), antibody A01289-1, controls and protocol steps. Open the full BRD7 IHC guide →

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BRD7 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	Nuclear staining in tissue (HPA tissue IHC); chromosome association (UniProt)
	Staining pattern	Broad nuclear staining across tissue cell types (HPA tissue IHC)
	Antigen retrieval	Citrate pH 6 HIER, heat-mediated (datasheet A01289-1)
	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 fixation conditions consistent across sections. (standard IHC practice; not target-specific)
	Caveat	Adipocytes may show no detectable staining (HPA tissue IHC)
	Regulation	Low tissue specificity (HPA tissue RNA)
	Isoform / epitope	Two isoforms; check antibody epitope coverage (UniProt)

Section 1

Recommended BRD7 IHC & IF Protocols

Start with the catalog antibody’s IHC-P protocol (datasheet: A01289-1), then compare four published BRD7 IHC protocols (PMC4102794; PMC4941311; PMC5938698; PMC10292652).

Recommended immunohistochemistry (IHC-P) protocol parameters
Sample	Paraffin-embedded human colon cancer tissue; fixative not specified (datasheet A01289-1)
Fixation	Image fixative and duration unreported (datasheet A01289-1); 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 A01289-1)
Peroxidase block	3% H2O2, 10 min, room temperature (standard)
Blocking	10% goat serum (datasheet A01289-1)
Primary antibody	Rabbit anti-BRD7, 0.5-1μg/ml (datasheet A01289-1)
Primary incubation	Overnight at 4 °C (datasheet A01289-1)
Detection	Streptavidin-biotin complex (SABC), DAB chromogen (datasheet A01289-1)
Counterstain	Hematoxylin, blue, dehydrate and mount (standard)
Expected result	BRD7-positive staining in glandular cells of adrenal gland (HPA tissue IHC: High). HPA tissue profile: Ubiquitous nuclear expression. No signal in the no-primary control.

💡Decision noteTry heat-mediated citrate retrieval at pH 6 first (datasheet: A01289-1); the breast cancer protocol also used citrate at pH 6 (PMC5938698).

Section 2

What Is the Expected BRD7 Staining Pattern?

BRD7 should stain predominantly in nuclei across many cell types (HPA: ubiquitous nuclear expression; Enhanced tissue IHC reliability). Glandular cells in colon and breast, respiratory epithelial cells in bronchus, and hematopoietic cells in bone marrow are reported as High (HPA: tissue IHC). This distribution fits a nuclear, chromosome-associated protein with no transmembrane segment (UniProt Q9NPI1: localization and topology).

What am I looking at on my slide?
Distinct nuclear staining in colon glandular cells or bone marrow hematopoietic cells.	This matches the expected compartment and reported High staining in those cells (HPA: tissue IHC). Judge the signal against neighboring nuclei and the section background; staining strength alone does not establish specificity (general IHC practice).
Cytoplasmic or surface staining dominates while nuclei remain faint.	This is discordant with the nuclear IHC profile (HPA: tissue IHC; UniProt Q9NPI1: nucleus, chromosome). Check the control section and detection background before interpreting it as BRD7. HPA also reports uncertain additional cytosolic localization by ICC-IF, so a faint cytoplasmic component is less decisive than a dominant one (HPA: subcellular ICC-IF).
Strong staining appears in adipocytes while expected nuclear staining is absent elsewhere.	Adipocytes are reported as Not detected (HPA: adipose tissue IHC). Treat this pattern as suspicious for nonspecific binding or endogenous detection activity, particularly if it persists in a no-primary control (general IHC practice). Do not use adipocytes as a universal negative control for every preparation.
Color spreads across tissue or obscures nuclear boundaries.	Diffuse background makes localization and intensity unreliable (general IHC practice). Examine a no-primary control and the pattern outside cells; widespread color can arise from nonspecific detection or insufficient blocking or washing (general IHC practice). It does not demonstrate the nuclear pattern reported by HPA (HPA: tissue IHC).
No nuclear signal appears in an otherwise interpretable positive-control section.	Colon glandular cells and bone marrow hematopoietic cells are reported as High (HPA: tissue IHC). A blank result there calls for checking antibody application, detection reagents and run controls (general IHC practice). It does not, by itself, establish BRD7 absence or a BRD7-specific fixation effect.

💡Expected BRD7 appearanceCall a result positive when distinct nuclear chromogen is visible in the expected cells, with readily detectable staining in reported High cells (HPA: tissue IHC); dominant cytoplasmic color or strong adipocyte staining without the expected nuclear pattern is suspect (HPA: tissue IHC; general IHC practice).

How each factor affects the staining
Tissue choice and validation	HPA reports ubiquitous nuclear expression, low tissue RNA specificity, and Enhanced tissue IHC reliability (HPA: tissue IHC). Its IHC result for CAB046458 is Enhanced (HPA: antibody validation). These findings support using a reported High tissue as a run control, while leaving local assay performance to be checked (general IHC practice).
Cell-level contrast	Colon glandular cells are High, hippocampal glial cells are Low, and adipocytes are Not detected (HPA: tissue IHC). Compare the named cell populations rather than assigning one intensity to every cell in a mixed section (general IHC practice).
Epitope and isoforms	UniProt lists 2 BRD7 isoforms and a bromodomain at residues 131–235 (UniProt Q9NPI1). No antibody epitope is supplied here; isoform coverage and effects of antigen retrieval on this antibody cannot be inferred from those annotations.
Topology and processing	BRD7 has no transmembrane segment or signal peptide and is annotated as a single 1–651 chain (UniProt Q9NPI1). Nuclear staining is biologically consistent with this record; these annotations provide no basis to predict shedding or a BRD7-specific fixation response.
Chromogenic detection background	If the assay uses peroxidase, endogenous peroxidase activity can contribute color; blocking and a no-primary control help assess it (general IHC practice). If another detection chemistry is used, assess background appropriate to that chemistry. HPA staining levels do not identify its source in a local assay.
IF/ICC Q: what localization should an IF result show?	A: Mainly nucleoplasmic signal is supported; additional cytosolic localization is uncertain (HPA: subcellular ICC-IF). This IF observation can inform compartment interpretation, but it is separate from the paraffin-section chromogenic IHC result (HPA: tissue IHC and subcellular ICC-IF).

Why is my staining missing, weak or wrong?
Situation	Likely cause	Next action
Positive-control nuclei are blank.	The run may have failed at antibody, retrieval or detection steps (general IHC practice); the image alone cannot identify which step failed.	Check run controls, reagent application and the assay's established retrieval conditions, then repeat the control section (general IHC practice). Use a reported High cell population, such as colon glandular cells (HPA: tissue IHC).
Nuclei stain, but background makes them hard to score.	Nonspecific binding, incomplete washing or detection background may reduce contrast (general IHC practice).	Review the no-primary control; assess blocking, antibody concentration and wash conditions within the established assay (general IHC practice). Score only nuclei with boundaries that remain interpretable.
Cytoplasmic color is stronger than nuclear color.	The result conflicts with the predominant nuclear IHC pattern (HPA: tissue IHC). Nonspecific staining or detection background is possible (general IHC practice).	Compare with a reported High nuclear control and a no-primary control (HPA: tissue IHC; general IHC practice). Record any cytoplasmic component separately; HPA's ICC-IF cytosolic assignment is uncertain (HPA: subcellular ICC-IF).
Adipocytes show strong color.	This conflicts with their Not detected HPA result and may reflect nonspecific or endogenous signal (HPA: adipose tissue IHC; general IHC practice).	Inspect the no-primary control and whether color occupies nuclei or tissue background; review blocking and detection controls before calling it BRD7 (general IHC practice).
One cell population looks weaker than another.	HPA reports different levels by cell type, including Low in hippocampal glial cells and High in cerebellar granular-layer cells (HPA: tissue IHC).	Identify the stained cell type before comparing intensity. Use the HPA cell-level observations as context, then judge the local run with its controls (HPA: tissue IHC; general IHC practice).
A staining difference is attributed to fixation.	No BRD7-specific fixation sensitivity is documented in the supplied UniProt or HPA records; staining intensity alone cannot establish that mechanism.	Document fixation and processing conditions, check run controls, and compare sections processed under the established assay conditions (general IHC practice). Report the observed difference without assigning a BRD7-specific fixation cause.

Source: UniProt Q9NPI1 Human Protein Atlas tissue Human Protein Atlas subcellular

Experiment evidence

IHC & IF images showing BRD7 staining

IHC analysis of BRD7 using anti-BRD7 antibody (A01289-1). BRD7 was detected in paraffin-embedded section of human colon cancer 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-BRD7 Antibody (A01289-1) 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.

A01289-1 · IHCIHC analysis of BRD7 using anti-BRD7 antibody (A01289-1). BRD7 was detected in paraffin-embedded section of human colon cancer tissue. Heat

IHC analysis of BRD7 using anti-BRD7 antibody (A01289-1). BRD7 was detected in paraffin-embedded section of human lung cancer 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-BRD7 Antibody (A01289-1) 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.

A01289-1 · IHCIHC analysis of BRD7 using anti-BRD7 antibody (A01289-1). BRD7 was detected in paraffin-embedded section of human lung cancer tissue. Heat m

HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000166164-BRD7/tissue/cerebral+cortex#imid_12660405

HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000166164-BRD7/tissue/colon#imid_12618939

HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000166164-BRD7/tissue/liver#imid_12618940

HPA · KidneyHuman Protein Atlas IHC, Kidney. Source: https://www.proteinatlas.org/ENSG00000166164-BRD7/tissue/kidney#imid_12618957

HPA · ICC-IF CACO-2Human Protein Atlas immunofluorescence, CACO-2, antibody HPA042389; green = BRD7, blue = nucleus, red = microtubules.

← swipe / scroll for more images →

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

Sample controls for BRD7 IHC & IF

🧪Run breast first: its glandular cells should stain (HPA: High in breast glandular cells). Use adipose tissue as the negative tissue (HPA: Not detected in adipocytes); adipocytes on the breast slide, if present, should lack specific staining and provide an internal negative comparison (HPA: Not detected in adipocytes).

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 BRD7 in CACO-2, Hep-G2, U2OS, HEK293, KOLF2.1J, with annotated localisation: Nucleoplasm (supported) (HPA subcellular).

Technical controls: Include a no-primary (secondary-only) control, a concentration-matched rabbit isotype control matched to the primary antibody’s clonality when known, and BRD7 knockout tissue or cells as a biological negative (caption: rabbit primary; standard IHC practice). Block endogenous peroxidase and assess endogenous biotin because the caption uses biotin-based detection with DAB (selected-SKU caption: biotinylated secondary, SABC and DAB).

⚠️Feasibility: A target-specific fixation window and fixation effect are unreported, and the selected-SKU caption does not state the fixative (selected-SKU caption: fixative not stated). The demonstrated paraffin-section procedure uses heat retrieval in citrate buffer at pH 6 for 20 minutes; whether retrieval is required for other preparations is unreported (selected-SKU caption: citrate retrieval). Frozen sections or IF cannot be judged easier from the supplied evidence; breast adipocytes may provide an unstained comparison, while biotin-based detection requires attention to background (HPA: Not detected in adipocytes; selected-SKU caption: SABC).

HPA tissue IHC evidence for BRD7

Comprehensive Human Protein Atlas IHC scoring per tissue (reliability: Enhanced — High consistency between antibody staining and RNA expression data.). Rows are taken directly from the HPA tissue chart — click any row's HPA link to view the source.

Positive expression · recommended positive controls

Tissue	Cell type	Level	Evidence	Source
Adrenal gland	Glandular cells	High	Protein (IHC)	HPA →
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
Adipose tissue	Adipocytes	Not detected	Protein (IHC)	HPA →

Section 3

Advanced BRD7 IHC Tips

Troubleshoot BRD7 chromogenic IHC by checking retrieval, nuclear localisation, controls, and scoring; IF/ICC considerations are covered separately below.

How should I retrieve BRD7 in paraffin sections when nuclear staining is weak?

Start with heat-mediated retrieval in citrate buffer at pH 6 for 20 minutes (datasheet A01289-1). The selected antibody produced staining in a paraffin-embedded human colon cancer section after this treatment, followed by 10% goat serum blocking and overnight incubation at 4°C with 1 μg/ml primary antibody (datasheet A01289-1). Compare retrieval runs on adjacent sections using the same detection and development conditions, so differences in nuclear signal are interpretable (standard IHC practice). If staining remains weak, check section adhesion, heating consistency, and reagent activity before changing retrieval conditions (standard IHC practice).

Can fixation explain weak or variable BRD7 staining?

BRD7-specific sensitivity to fixation is unknown because the selected tissue-IHC caption does not state a fixative (datasheet A01289-1). Its paraffin-section result therefore supports the reported retrieval and staining workflow, but cannot establish which fixation duration preserves the epitope (datasheet A01289-1). Record the fixative and fixation time for each specimen, then compare matched sections processed together with citrate pH 6 retrieval for 20 minutes (datasheet A01289-1; standard IHC practice). If a batch loses nuclear staining, review fixation records alongside retrieval, primary incubation, and control-section performance before assigning the cause (standard IHC practice).

What staining compartment should I expect for BRD7?

Expect predominantly nuclear staining in tissue: BRD7 is annotated in the nucleus and on chromosomes, and tissue IHC reports ubiquitous nuclear expression (UniProt Q9NPI1 localisation; HPA tissue IHC profile). Supported ICC/IF localisation is mainly nucleoplasmic, while additional cytosolic localisation is uncertain, so isolated strong cytoplasmic DAB deserves scrutiny (HPA subcellular). Inspect nuclear staining in glandular cells of colon or appendix as contextual comparators, where HPA reports high expression (HPA: High in colon glandular cells; HPA: High in appendix glandular cells). Score nuclei separately from cytoplasm and assess morphology before calling a divergent pattern positive (standard IHC practice).

Could isoforms or epitope masking alter my BRD7 IHC result?

BRD7 has 2 annotated isoforms, but the supplied evidence does not map this antibody's epitope to either one (UniProt Q9NPI1 isoforms; datasheet A01289-1). Its bromodomain spans residues 131–235, and annotated modified residues include phosphoserines and an acetyllysine; these annotations alone do not establish antibody sensitivity to modification (UniProt Q9NPI1 domains and modified residues). Keep retrieval at citrate pH 6 for 20 minutes when comparing specimens, because that is the documented condition for this antibody (datasheet A01289-1). If staining differs, seek an epitope map or orthogonal expression evidence before attributing the difference to an isoform or modification (standard IHC practice).

How should I assess BRD7 in a multiplex IF experiment?

Plan to assess BRD7 primarily in nuclei, since its supported subcellular location is nucleoplasmic; the reported additional cytosolic location is uncertain (HPA subcellular). Pair the BRD7 channel with a validated marker for the expected cell type and a nuclear counterstain, then inspect whether signals occupy the same cells (HPA tissue IHC profile; standard IF practice). Choose spectrally separated fluorophores after checking tissue autofluorescence and include single-channel controls for bleed-through (standard IF practice). Because BRD7 lacks a transmembrane segment and is nuclear, use permeabilisation that gives antibodies access to intracellular nuclear epitopes, then verify morphology and background experimentally (UniProt Q9NPI1 topology and localisation; standard IF practice).

How can I reduce diffuse or misleading DAB background?

Separate diffuse deposit from nuclear signal by inspecting morphology and a section processed without primary antibody (standard IHC practice; HPA tissue IHC profile). The selected paraffin-section workflow used 10% goat serum, a biotinylated secondary antibody, streptavidin–biotin detection, and DAB; these details identify points where nonspecific staining or detection background should be checked (datasheet A01289-1; standard IHC practice). Include an appropriate peroxidase block before chromogen development and keep wash and development conditions consistent between controls and test sections (standard IHC practice). If background persists, review secondary-only staining and endogenous biotin interference in this biotin-based workflow (datasheet A01289-1; standard IHC practice).

What is a defensible way to quantify BRD7 IHC? ⚠ ANSWER MARKED FOR VERIFICATION

Define the cell population and score BRD7 nuclear staining, consistent with its reported tissue pattern and supported nucleoplasmic localisation (HPA tissue IHC profile; HPA subcellular). For a graded readout, use an H-score from the percentage of nuclei at each intensity; report the percentage of positive nuclei separately if intensity thresholds are uncertain (standard IHC practice). Normalise counts to the number of evaluable nuclei in the same annotated compartment, or report positive nuclei per mm² when cell density itself matters (standard IHC practice). Keep retrieval, imaging, chromogen development, and scoring thresholds consistent, and exclude necrotic or damaged regions by a prespecified rule (standard IHC practice).

When is an unexpected BRD7 signal likely to be artefactual?

A convincing BRD7 result should show interpretable nuclear staining in intact cells, matching the reported ubiquitous nuclear tissue pattern and supported nucleoplasmic localisation (HPA tissue IHC profile; HPA subcellular). High staining in colon glandular cells can provide context, while adipocytes are reported as not detected; neither observation alone proves specificity in a new specimen (HPA: High in colon glandular cells; HPA: Not detected in adipocytes; standard IHC practice). Treat staining confined to section edges, necrotic regions, or the wrong cell compartment cautiously (standard IHC practice; UniProt Q9NPI1 localisation). Compare primary-omission controls and peroxidase-blocked sections to distinguish antibody-dependent nuclear signal from detection artefact (standard IHC practice).

Boster reagents

Best BRD7 / Bromodomain-containing protein 7 IHC Antibodies

A01289-1 has IHC images from human colon and lung cancer paraffin sections (catalog image captions); its listed reactivity covers human, mouse and rat (catalog applications and reactivity).

Real IHC data

Anti-BRD7 Antibody ®

Cat # A01289-1

View product →

A01289-1 will render with an IHC image of a human colon cancer paraffin section (catalog image caption). A second catalog caption shows IHC in a human lung cancer paraffin section; IF/ICC data are unreported (catalog image captions and applications).

Which to pick: Choose A01289-1 for paraffin section IHC: its caption documents citrate retrieval at pH 6 for 20 minutes and a primary concentration of 1 μg/mL (A01289-1 IHC image caption); the fixative is unreported (A01289-1 IHC image caption). Its listed human, mouse and rat reactivity makes it the cross-species candidate, although the supplied IHC images show human tissue only (catalog reactivity and image captions). No IF/ICC antibody is supported by this payload because A01289-1 has no listed IF/ICC application or IF image (catalog applications and image data).

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

References

UniProt Consortium. UniProt entry Q9NPI1 (BRD7_HUMAN, Bromodomain-containing protein 7).
Human Protein Atlas. BRD7 tissue IHC expression (reliability: Enhanced).
Human Protein Atlas. BRD7 subcellular location (ICC-IF): Mainly localized to the nucleoplasm. In addition localized to the cytosol..
Human Protein Atlas. BRD7 antibody validation summary (3 antibodies).
Targeting the anaphase-promoting complex/cyclosome (APC/C)- bromodomain containing 7 (BRD7) pathway for human osteosarcoma. Oncotarget 2014 — PMC4102794.
Bromodomain-containing protein 7 (BRD7) as a potential tumor suppressor in hepatocellular carcinoma. Oncotarget 2016 — PMC4941311.
BRD7 inhibits the Warburg effect and tumor progression through inactivation of HIF1α/LDHA axis in breast cancer. Cell death & disease 2018 — PMC5938698.
Machine learning identifies ferroptosis-related genes as potential diagnostic biomarkers for osteoarthritis. Frontiers in endocrinology 2023 — PMC10292652.
PubMed PMID:11025449 — UniProt-cited evidence.
PubMed PMID:12489984 — UniProt-cited evidence.
PubMed PMID:15616553 — UniProt-cited evidence.