BCL10 / B-cell lymphoma/leukemia 10 · IHC design guide

Design Immunohistochemistry for BCL10

Plan BCL10 chromogenic IHC around the cytoplasmic staining seen in most cell types (HPA tissue IHC). This guide covers fixation, retrieval, detection and interpretation, including the possibility of nuclear staining in some tumor cells (UniProt).

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

Immunohistochemistry protocol sheet for BCL10 (IHC for BCL10): expected localisation Cytoplasmic staining in most cell types (HPA tissue IHC), antibody PB9525, validated IHC image, and IHC protocol steps — Printable **BCL10** IHC protocol sheet — expected localisation Cytoplasmic staining in most cell types (HPA tissue IHC), antibody PB9525, controls and protocol steps. Open the full BCL10 IHC guide →

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BCL10 Immunohistochemistry Experimental Design Guide

Expected localisation, validated protocols, controls and antibodies — the at-a-glance facts below, then the full design guide.

Must know before staining

	Expected localisation	Cytoplasmic staining in most cell types (HPA tissue IHC)
	Staining pattern	Cytoplasmic signal in most cell types (HPA tissue IHC)
	Antigen retrieval	EDTA pH 8.0 HIER, heat-mediated (datasheet PB9525)
	Positive control ⓘ	Adrenal gland+4 more · see all
	Negative control ⓘ	Adipose tissue+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)
	Caveat	Some tumor cells may show nuclear BCL10 (UniProt)
	Regulation	Ubiquitous tissue expression (UniProt)
	Isoform / epitope	No annotated isoforms; one 1–233 chain (UniProt)

Section 1

Recommended BCL10 IHC & IF Protocols

Start with the catalog antibody’s EDTA pH 8.0 protocol (datasheet: PB9525); the four published IHC protocols below provide tissue specific comparisons (PMC5662601; PMC8375138; PMC12421401; PMC9312497).

Recommended immunohistochemistry (IHC-P) protocol parameters
Sample	Paraffin-embedded human lymph node carcinoma tissue; fixative not specified (datasheet PB9525)
Fixation	Image fixative and duration unreported (datasheet PB9525); 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 PB9525); 20 min, 95–100 °C (standard)
Peroxidase block	3% H2O2, 10 min, room temperature (standard)
Blocking	10% goat serum (datasheet PB9525)
Primary antibody	Rabbit anti-BCL10, 2-5 μg/ml (datasheet PB9525)
Primary incubation	Overnight at 4 °C (datasheet PB9525)
Detection	HRP-conjugated secondary, DAB chromogen (datasheet PB9525)
Counterstain	Hematoxylin, blue, dehydrate and mount (standard)
Expected result	BCL10-positive staining in glandular cells of adrenal gland (HPA tissue IHC: Medium). HPA tissue profile: Cytoplasmic expression in most cell types. No signal in the no-primary control.

💡Decision noteTry EDTA pH 8.0 first for the catalog antibody (datasheet: PB9525). If optimizing retrieval, published alternatives include citrate pH 6.0 and Tris EDTA pH 9.0 (PMC12421401; PMC9312497).

Section 2

What Is the Expected BCL10 Staining Pattern?

BCL10 should stain mainly in the cytoplasm, sometimes with a compact perinuclear pattern; it has no transmembrane segment (UniProt O95999, subcellular location and topology). HPA reports cytoplasmic expression in most cell types and medium staining in selected glandular and epithelial cells (HPA tissue IHC). Its tissue IHC reliability is Approved, with medium consistency between staining and RNA expression (HPA tissue IHC).

What am I looking at on my slide?
Cytoplasmic staining in appendix glandular cells or bronchial respiratory epithelial cells, with identifiable cell borders.	This matches the reported medium staining in those cells (HPA tissue IHC) and the principal cytoplasmic location (UniProt O95999). Score staining in the named cells; surrounding unstained cells do not invalidate a positive result (general IHC practice).
Predominantly nuclear staining in ordinary tissue, with little or no cytoplasmic staining.	Review as a possible localisation artefact because the expected tissue pattern is cytoplasmic (HPA tissue IHC). Nuclear signal alone is not conclusive: UniProt reports BCL10 in nuclei of several tumor cell types, and HPA reports an additional nucleoplasmic ICC-IF location (UniProt O95999; HPA subcellular).
Strong staining in adipocytes or cardiomyocytes while the expected positive cells are weak.	HPA lists adipocytes in adipose tissue and cardiomyocytes in heart muscle as not detected (HPA tissue IHC). Treat discordant staining as possible cross-reactivity or endogenous detection activity (general IHC practice); first check whether the signal persists when primary antibody is omitted.
Uniform chromogen haze across cells, extracellular areas and the slide background.	This distribution does not resemble the reported cell-associated cytoplasmic pattern (HPA tissue IHC). Consider nonspecific reagent binding, inadequate washing or endogenous detection activity (general IHC practice); a stained region alone does not establish BCL10 localisation.
No staining in appendix glandular cells on a run intended to include a positive control.	Those cells have reported medium staining (HPA tissue IHC). An absent control signal makes a negative study section difficult to interpret (general IHC practice). Check section integrity and the assay steps before calling the study specimen BCL10-negative.

💡Expected BCL10 appearanceCall a positive result when identifiable glandular or epithelial cells show predominantly cytoplasmic, approximately medium staining in a reported positive tissue (HPA tissue IHC); strong signal confined to HPA-listed undetected cells or diffuse slide haze warrants an artefact check (HPA tissue IHC; general IHC practice).

How each factor affects the staining
Cell and compartment choice	Use a named HPA-positive cell population when judging localisation: appendix glandular cells and bronchial respiratory epithelial cells are medium, whereas adipocytes in adipose tissue are not detected (HPA tissue IHC). The tissue label alone is less precise than the cell label.
Perinuclear pattern and assembly	A compact or filamentous perinuclear appearance is described for BCL10 (UniProt O95999, subcellular location). UniProt also describes CARD-dependent BCL10 oligomerization (UniProt O95999, subunit); morphology alone cannot establish pathway activation (general IHC interpretation).
Antibody evidence	HPA lists IHC as Approved for HPA017925 and CAB001944; it lists ICC as Supported for HPA018953 (HPA antibodies). These are application-specific statuses, and the supplied record does not establish that an ICC-tested antibody performs in paraffin IHC.
Strength of tissue evidence	HPA describes the tissue IHC profile as cytoplasmic in most cell types and rates its agreement with RNA data as medium (HPA tissue IHC). Use the reported levels as comparison points, not absolute thresholds for every specimen (general IHC interpretation).
Detection system	Chromogenic signal can arise from endogenous tissue activity or nonspecific reagent binding (general IHC practice). Primary-omission and detection-only controls help separate those signals from antibody-dependent staining; neither control establishes molecular specificity by itself (general IHC practice).

Why is my staining missing, weak or wrong?
Situation	Likely cause	Next action
Positive-control tissue is blank.	A missed assay step, weak detection or unsuitable control section can produce no signal (general IHC practice).	Repeat with a documented positive cell population, such as appendix glandular cells with reported medium staining (HPA tissue IHC). Verify retrieval and detection steps against the IHC-validated antibody's instructions (general IHC practice).
Positive cells show diffuse nuclear signal without the expected cytoplasm.	The distribution differs from the principal tissue pattern (HPA tissue IHC), though nuclear localisation has been reported in some tumor cells (UniProt O95999).	Compare cytoplasmic and nuclear signal in a known-positive control, inspect morphology and run a primary-omission control (general IHC practice). Record a genuine nuclear component separately instead of forcing a cytoplasmic score.
Adipocytes stain strongly.	Adipocytes in adipose tissue are listed as not detected (HPA tissue IHC); cross-reactivity or endogenous detection activity is possible (general IHC practice).	Check a primary-omission control and the detection system, then compare staining in a named HPA-positive population (general IHC practice; HPA tissue IHC). Do not use adipocyte colour alone as evidence of BCL10 expression.
Background obscures cell boundaries.	Nonspecific binding, residual detection reagents or inadequate washing can obscure a cell-associated pattern (general IHC practice).	Inspect an unstained section and a primary-omission control; review blocking, washing and chromogen development for the chosen detection system (general IHC practice). Score only where cell boundaries and cytoplasmic signal remain interpretable.
A low-staining cell population appears negative.	HPA reports low staining in cerebral cortex neuronal cells and pancreatic exocrine glandular cells (HPA tissue IHC). Weak signal may be difficult to distinguish from assay background (general IHC practice).	Interpret alongside a reported medium positive cell population and a negative control (HPA tissue IHC; general IHC practice). Avoid declaring assay failure from a low-staining population alone.
IF/ICC Q: Should nucleoplasmic fluorescence replace the IHC expectation?	HPA reports cytosol as the main ICC-IF location and nucleoplasm as an additional location (HPA subcellular); its tissue IHC profile is cytoplasmic (HPA tissue IHC).	A: Interpret IF/ICC on its own guide page and record both compartments when visible (HPA subcellular). For paraffin IHC, judge the tissue pattern against the cytoplasmic IHC evidence (HPA tissue IHC).

Source: UniProt O95999 Human Protein Atlas tissue Human Protein Atlas subcellular

Experiment evidence

IHC & IF images showing BCL10 staining

IHC analysis of Bcl10 using anti-Bcl10 antibody (PB9525). Bcl10 was detected in a paraffin-embedded section of human lymph node carcinoma tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH 8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 2 μg/ml rabbit anti-Bcl10 Antibody (PB9525) overnight at 4°C. Peroxidase Conjugated Goat Anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using HRP Conjugated Rabbit IgG Super Vision Assay Kit (Catalog # SV0002) with DAB as the chromogen.

PB9525 · IHCIHC analysis of Bcl10 using anti-Bcl10 antibody (PB9525). Bcl10 was detected in a paraffin-embedded section of human lymph node carcinoma ti

Immunohistochemical analysis of paraffin-embedded H. kidney section using BCL10 Antibody. M01616 was diluted at 1:25 dilution. A peroxidase-conjugated goat anti-mouse IgG at 1:400 dilution was used as the secondary antibody, followed by DAB staining.

M01616 · IHCImmunohistochemical analysis of paraffin-embedded H. kidney section using BCL10 Antibody. M01616 was diluted at 1:25 dilution. A peroxidase-

Immunohistochemical analysis of paraffin-embedded M. kidney section using BCL10 Antibody. M01616 was diluted at 1:25 dilution. A peroxidase-conjugated goat anti-mouse IgG at 1:400 dilution was used as the secondary antibody, followed by DAB staining.

M01616 · IHCImmunohistochemical analysis of paraffin-embedded M. kidney section using BCL10 Antibody. M01616 was diluted at 1:25 dilution. A peroxidase-

M01616-1 · IHCImmunohistochemical analysis of paraffin-embedded human breast cancer, using Bcl10 Antibody.

A01616 · IFImmunocytochemistry of Bcl10 in Raji cells with Bcl10 antibody at 1 μg/mL.

HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000142867-BCL10/tissue/cerebral+cortex#imid_635376

HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000142867-BCL10/tissue/colon#imid_635318

HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000142867-BCL10/tissue/liver#imid_635303

HPA · KidneyHuman Protein Atlas IHC, Kidney. Source: https://www.proteinatlas.org/ENSG00000142867-BCL10/tissue/kidney#imid_635312

HPA · ICC-IF A-431Human Protein Atlas immunofluorescence, A-431, antibody HPA018953; green = BCL10, blue = nucleus, red = microtubules.

← swipe / scroll for more images →

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

Sample controls for BCL10 IHC & IF

🧪Run adrenal gland first and score its glandular cells for BCL10 staining (HPA: Medium in adrenal gland glandular cells). Use adipose tissue adipocytes as the negative comparison (HPA: Not detected in adipose tissue adipocytes); on the adrenal slide, assess staining in adjacent non-glandular cells as background without assuming they are BCL10-negative (UniProt O95999: ubiquitous tissue specificity).

Positive control tissue: Adrenal gland (Glandular cells, HPA Medium)

Negative control tissue: Adipose tissue (HPA Not detected)

ICC-IF cell lines (HPA subcellular resource): HPA ICC-IF images show BCL10 in A-431, GAMG, U2OS, with annotated localisation: Cytosol (supported) (HPA subcellular).

Technical controls: Include a no-primary, secondary-only control; nonimmune rabbit IgG matched to the primary antibody’s isotype and clonality where known; and BCL10-knockout material processed in parallel (selected-SKU IHC caption: rabbit primary antibody; standard IHC practice). Quench endogenous peroxidase before HRP–DAB detection and check adrenal pigment against the no-primary slide (selected-SKU IHC caption: HRP–DAB detection; standard IHC practice).

⚠️Feasibility: The selected PB9525 paraffin-section caption reports EDTA retrieval at pH 8.0, but its fixative is unreported; no target-specific fixation window or fixation effect is supplied (selected-SKU IHC caption). Retrieval is documented for that example, but its necessity for every specimen is unestablished (selected-SKU IHC caption). ICC–IF images exist, while matched frozen-section evidence is absent, so neither frozen sections nor IF can be judged easier from these payloads (HPA subcellular: A-431, GAMG and U2OS ICC–IF images; selected-SKU IHC caption: paraffin section).

HPA tissue IHC evidence for BCL10

Comprehensive Human Protein Atlas IHC scoring per tissue (reliability: Approved — Medium 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	Medium	Protein (IHC)	HPA →
Appendix	Glandular cells	Medium	Protein (IHC)	HPA →
Bronchus	Respiratory epithelial cells	Medium	Protein (IHC)	HPA →
Cervix	Glandular cells	Medium	Protein (IHC)	HPA →
Colon	Endothelial cells	Medium	Protein (IHC)	HPA →

Undetected expression · recommended negative controls

Tissue	Cell type	Level	Evidence	Source
Adipose tissue	Adipocytes	Not detected	Protein (IHC)	HPA →
Bone marrow	Hematopoietic cells	Not detected	Protein (IHC)	HPA →
Breast	Adipocytes	Not detected	Protein (IHC)	HPA →
Caudate	Glial cells	Not detected	Protein (IHC)	HPA →
Cerebellum	Cells in granular layer	Not detected	Protein (IHC)	HPA →

Section 3

Advanced BCL10 IHC Tips

Troubleshoot BCL10 staining in paraffin sections by checking retrieval, compartment, controls and cell-specific scoring before interpreting chromogenic signal.

Which retrieval conditions should I try first for weak BCL10 staining?

Start with heat-mediated antigen retrieval in EDTA at pH 8.0 for paraffin sections (datasheet PB9525). The documented BCL10 image used that condition, followed by 10% goat serum blocking and 2 μg/ml primary antibody overnight at 4°C (caption PB9525). If staining remains weak, compare a shorter and longer heating interval on adjacent sections while holding antibody concentration and detection constant (standard IHC practice). Assess whether cytoplasmic staining improves in glandular cells before accepting stronger diffuse signal as improvement (HPA: cytoplasmic expression; HPA: medium staining in several glandular cell types). Keep retrieval conditions identical across samples intended for quantitative comparison (standard IHC practice).

How should I troubleshoot fixation-related loss of BCL10 signal?

Target-specific sensitivity of BCL10 to fixation is unknown from the supplied evidence; the selected paraffin-section caption does not state a fixative (caption PB9525). Record fixative, fixation duration, tissue thickness and processing history for each specimen before attributing weak staining to fixation (standard IHC practice). Compare similarly processed sections using the documented EDTA retrieval at pH 8.0 and 2 μg/ml primary antibody conditions (caption PB9525). Include a reference section that previously gave interpretable cytoplasmic staining in each run, and inspect tissue preservation alongside stain intensity (standard IHC practice; HPA: cytoplasmic expression). Avoid inferring fixation sensitivity from BCL10 localisation or phosphorylation alone (UniProt O95999: localisation and modified residues).

Should BCL10 staining be cytoplasmic, nuclear or membrane-associated?

Prioritise cytoplasmic staining when assessing paraffin sections because that is the predominant tissue pattern reported for BCL10 (HPA: cytoplasmic expression in most cell types). Perinuclear compact or filamentous staining is plausible, and nuclear staining has been observed in several tumor cell types (UniProt O95999: subcellular localisation). BCL10 can occur in membrane rafts but has no transmembrane segment, so a continuous cell-surface rim needs careful validation (UniProt O95999: localisation and topology). Compare suspected nuclear signal with a counterstain and adjacent cytoplasm at the same magnification (standard IHC practice). Record compartments separately rather than combining all brown signal into one positive score (standard IHC practice).

Could the epitope or protein processing explain inconsistent BCL10 IHC staining?

Check the antibody's stated immunogen or mapped epitope before assigning inconsistent staining to epitope loss; no epitope is supplied here (catalog evidence supplied). The BCL10 record lists 0 isoforms and a single chain spanning residues 1–233, so an isoform-specific staining explanation lacks support (UniProt O95999: isoforms and processing). Its CARD domain occupies residues 13–101, and serine 138 is a reported phosphorylation site (UniProt O95999: domains and modified residues). Neither observation establishes that fixation or retrieval changes this antibody's binding (UniProt O95999: domain and modified-residue annotations; caption PB9525: no mapped epitope). Compare retrieval-matched sections and appropriate controls before interpreting pattern differences as molecular variants (standard IHC practice).

How can I check BCL10 localisation by multiplex IF?

Treat IF as a separately optimised application because the supplied PB9525 tissue image documents chromogenic paraffin-section IHC (caption PB9525). Pair BCL10 with a validated marker identifying the expected cell population, such as glandular cells in a tissue where their staining is reported (HPA: medium glandular-cell staining in appendix and duodenum). Select fluorophores after checking unstained tissue autofluorescence, and favour a spectrally separated far-red channel when shorter wavelengths show substantial background (standard IF practice). Include permeabilisation suitable for a cytosolic, sometimes nucleoplasmic target, then compare with an omitted-primary control (HPA: cytosol and nucleoplasm; standard IF practice). Do not transfer the IHC dilution of 2 μg/ml directly into IF without titration (caption PB9525; standard IF practice).

What causes diffuse brown background in BCL10 sections?

First compare a no-primary control with the stained section to separate detection-system signal from primary-antibody-associated staining (standard IHC practice). Block endogenous peroxidase before HRP and DAB development, and review the effectiveness of the 10% goat serum block documented for PB9525 (standard IHC practice; caption PB9525). If background persists, titrate around the documented 2 μg/ml primary concentration while keeping retrieval at EDTA pH 8.0 (caption PB9525; standard IHC practice). Inspect folds, section edges and poorly preserved regions separately because local staining there can mislead interpretation (standard IHC practice). Judge any adjustment against expected cytoplasmic signal rather than overall section darkness (HPA: cytoplasmic expression; standard IHC practice).

How should I score heterogeneous BCL10 staining in paraffin sections? ⚠ ANSWER MARKED FOR VERIFICATION

Define the cell population and compartment before scoring; BCL10 tissue staining is mainly cytoplasmic, while nuclear localisation can occur in some tumors (HPA: cytoplasmic expression; UniProt O95999: tumor-cell nuclear localisation). For cytoplasmic chromogen, record the percentage of cells at each intensity and calculate an H-score from 0–300, or report percentage positive using a prespecified threshold (standard IHC practice). If cells are sparse, report positive-cell density per mm² of viable tissue and state the counted area (standard IHC practice). Normalise counts to eligible cells or viable tissue area, not total image area containing empty space (standard IHC practice). Apply one retrieval, detection and imaging protocol across compared sections (standard IHC practice).

How do I distinguish true BCL10 signal from artefact?

Look for reproducible cytoplasmic staining in intact cells, with perinuclear or filamentous patterns considered in context (HPA: cytoplasmic expression; UniProt O95999: perinuclear pattern). A strong cell-surface rim is suspect without corroboration because BCL10 has no transmembrane segment (UniProt O95999: topology). Compare staining with an omitted-primary section and review edges, folds, necrotic areas and residual endogenous peroxidase signal before calling a specimen positive (standard IHC practice). Interpret nuclear staining cautiously: it has been reported in several tumor cell types, but distribution alone does not establish specificity (UniProt O95999: tumor-cell nuclear localisation; standard IHC practice). Use cell identity and matched controls because HPA tissue staining has only medium consistency with RNA expression (HPA: Approved, medium consistency).

Boster reagents

Best BCL10 / B-cell lymphoma/leukemia 10 IHC Antibodies

Anti-BCL10 antibodies have IHC images from human paraffin tissue sections (PB9525, M01616, M01616-1: image captions) and IF/ICC images from Raji cells (A01616: image captions).

Real IHC data

Anti-Bcl10 Antibody ®

Cat # PB9525

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Real IHC data Immunohistochemical analysis of paraffin-embedded H. kidney section using BCL10 Antibody. M01616 was diluted at 1:25 dilution. A peroxidase-conjugated goat anti-mouse IgG at 1:400 dilution was used as the secondary antibody, followed by DAB staining.

Anti-BCL10 Antibody

Cat # M01616

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Real IHC data Immunohistochemical analysis of paraffin-embedded human breast cancer, using Bcl10 Antibody.

Anti-Bcl10 Rabbit Monoclonal Antibody

Cat # M01616-1

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Real IF data Immunocytochemistry of Bcl10 in Raji cells with Bcl10 antibody at 1 μg/mL.

Anti-Bcl-10 Antibody

Cat # A01616

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PB9525 shows human lymph node carcinoma IHC, while M01616 shows human kidney IHC (PB9525, M01616: image captions). M01616-1 shows human breast cancer IHC, and A01616 shows Raji cell ICC and IF (M01616-1, A01616: image captions).

Which to pick: For paraffin section IHC, choose PB9525 for its documented EDTA pH 8.0 retrieval and 2 μg/ml primary incubation (PB9525: image caption), or M01616 for its 1:25 human kidney example (M01616: image caption); M01616-1 also has a paraffin section image from human breast cancer (M01616-1: image caption). For IF/ICC, choose A01616 because it has Raji cell ICC and IF images; it also has the broadest listed species reactivity—human, mouse and rat (A01616: image captions and catalog applications/reactivity). The paraffin captions do not report a fixative (PB9525, M01616, M01616-1: image captions).

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

References

UniProt Consortium. UniProt entry O95999 (BCL10_HUMAN, B-cell lymphoma/leukemia 10).
Human Protein Atlas. BCL10 tissue IHC expression (reliability: Approved).
Human Protein Atlas. BCL10 subcellular location (ICC-IF): Mainly localized to the cytosol. In addition localized to the nucleoplasm..
Human Protein Atlas. BCL10 antibody validation summary (3 antibodies).
First-line antibiotic therapy in Helicobacter pylori-negative low-grade gastric mucosa-associated lymphoid tissue lymphoma. Scientific reports 2017 — PMC5662601.
Contribution of nuclear BCL10 expression to tumor progression and poor prognosis of advanced and/or metastatic pancreatic ductal adenocarcinoma by activating NF-κB-related signaling. Cancer cell international 2021 — PMC8375138.
B cell CLL/lymphoma 10 promotes colorectal cancer cell proliferation and regulates cuproptosis sensitivity through the NF-κB signaling pathway. World journal of gastroenterology 2025 — PMC12421401.
Bcl10 Regulates Lipopolysaccharide-Induced Pro-Fibrotic Signaling in Bronchial Fibroblasts from Severe Asthma Patients. Biomedicines 2022 — PMC9312497.
PubMed PMID:9989495 — UniProt-cited evidence.
PubMed PMID:10187770 — UniProt-cited evidence.
PubMed PMID:10187771 — UniProt-cited evidence.