BCL2L11 / Bcl-2-like protein 11 · IHC design guide

Design Immunohistochemistry for BCL2L11

Plan chromogenic BCL2L11 IHC in paraffin sections using the observed cytoplasmic pattern and documented positive cell populations (HPA tissue IHC). Compare the antibody epitope with the 20 annotated isoforms before interpreting staining intensity (UniProt).

Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
Immunohistochemistry protocol sheet for BCL2L11 (IHC for BCL2L11): expected localisation General cytoplasmic tissue staining (HPA tissue IHC), antibody A01552-4, validated IHC image, and IHC protocol steps
Printable BCL2L11 IHC protocol sheet — expected localisation General cytoplasmic tissue staining (HPA tissue IHC), antibody A01552-4, controls and protocol steps. Open the full BCL2L11 IHC guide →

BCL2L11 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 General cytoplasmic tissue staining (HPA tissue IHC)
Staining pattern Cytoplasmic staining; high in kidney tubules and lung macrophages (HPA tissue IHC)
Antigen retrieval EDTA pH 8.0 HIER, heat-mediated (datasheet A01552-4)
Positive control ⓘ Appendix+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 sections (standard IHC practice; not target-specific)
Caveat Isoform diversity may affect antibody epitope coverage (UniProt)
Regulation Phosphorylation-linked degradation may lower signal (UniProt)
Isoform / epitope 20 isoforms; verify epitope coverage (UniProt)
Section 1

Recommended BCL2L11 IHC & IF Protocols

The catalog antibody's IHC-P protocol (datasheet A01552-4) is presented alongside a published thyroid IHC protocol (PMC12881209).

Recommended immunohistochemistry (IHC-P) protocol parameters
SampleParaffin-embedded human lung cancer tissue; fixative not specified (datasheet A01552-4)
FixationImage fixative and duration unreported (datasheet A01552-4); verify before use.
Sectioning4–5 µm sections on charged slides (standard)
DeparaffinisationXylene, graded ethanol series to water (standard)
Antigen retrievalHeat retrieval: EDTA pH 8.0 (datasheet A01552-4); 20 min, 95–100 °C (standard)
Peroxidase block3% H2O2, 10 min, room temperature (standard)
Blocking10% goat serum (datasheet A01552-4)
Primary antibodyRabbit anti-BCL2L11, 2-5 μg/ml (datasheet A01552-4)
Primary incubationOvernight at 4 °C (datasheet A01552-4)
DetectionStreptavidin-biotin complex (SABC), DAB chromogen (datasheet A01552-4)
CounterstainHematoxylin, blue, dehydrate and mount (standard)
Expected resultBCL2L11-positive staining in lymphoid tissue of appendix (HPA tissue IHC: High). HPA tissue profile: General cytoplasmic expression. No signal in the no-primary control.
💡Decision noteStart with heat-mediated EDTA pH 8.0 retrieval for the catalog antibody (datasheet A01552-4). The thyroid study used CC1 pretreatment (PMC12881209).
Section 2

What Is the Expected BCL2L11 Staining Pattern?

BCL2L11 should appear chiefly in the cytoplasm on IHC, consistent with its association with intracytoplasmic membranes and mitochondria despite having no annotated transmembrane segment (HPA: general cytoplasmic IHC expression; UniProt O43521: subcellular location and topology). Strong staining is reported in kidney tubule cells, lung macrophages and endometrial glandular cells, among others (HPA: High in each). Treat intensity as a guide: HPA rates the tissue IHC profile Approved, with medium consistency between staining and RNA data (HPA: reliability).

What am I looking at on my slide?
Cytoplasmic staining in kidney tubule cells, lung macrophages or endometrial glandular cells.This matches reported High staining in those cell types (HPA: tissue IHC). Assess intensity against neighboring cells within the same section; a positive result need not outline individual mitochondria in chromogenic IHC (standard IHC interpretation).
Signal confined to nuclei, with little or no cytoplasmic staining.An isolated nuclear pattern conflicts with the reported cytoplasmic IHC profile and mitochondrial or intracytoplasmic membrane location (HPA: tissue IHC; UniProt O43521: subcellular location). Reassess staining specificity and distinguish chromogen from the nuclear counterstain (standard IHC practice).
Strong staining mainly in a cell population reported as Low, such as smooth muscle cells or fibroblasts.That distribution differs from the HPA tissue examples (HPA: Low in smooth muscle cells and soft-tissue fibroblasts). Consider cross-reactivity or endogenous detection activity before calling it BCL2L11; a Low designation does not mean every cell must be blank (standard IHC interpretation).
Diffuse color across cells, stroma or slide regions, without recognizable cellular boundaries.A widespread haze is difficult to score as the reported cytoplasmic pattern (HPA: tissue IHC). Uneven detection or nonspecific background is possible; compare a matched negative control and inspect whether the signal follows tissue structures (standard IHC practice).
No detectable staining in a kidney section containing tubules, despite an interpretable counterstain.Tubule cells are a reported High-staining reference, so an entirely blank result warrants a technical check (HPA: High in kidney tubule cells). It does not alone establish biological absence: evaluate tissue preservation, antibody and detection controls, and the scoring threshold (standard IHC practice).
💡Expected BCL2L11 appearanceCall positive when interpretable cytoplasmic staining is strong in an HPA High cell population, such as kidney tubule cells; isolated nuclear color or structureless haze is suspect (HPA: tissue IHC; standard IHC interpretation).
How each factor affects the staining
Compartment and resolutionUniProt places BCL2L11 at mitochondria and intracytoplasmic membranes, while HPA describes general cytoplasmic tissue IHC staining (UniProt O43521: subcellular location; HPA: tissue IHC). Score the compartment visible by chromogenic microscopy rather than requiring resolved mitochondrial puncta (standard IHC practice).
Tissue and isoform variationBimEL, BimL and BimS are widely expressed with tissue-specific variation; Bim-gamma is most abundant in small intestine and colon (UniProt O43521: tissue specificity). The HPA IHC levels describe observed cell populations, not isoform-specific staining (HPA: tissue IHC).
Strength of IHC evidenceHPA calls the tissue IHC profile Approved and reports medium agreement between antibody staining and RNA expression (HPA: reliability). CAB026332 is IHC Approved; no IHC Enhanced designation is supplied for it (HPA: antibody validation). Interpret unexpected staining with controls rather than treating the profile as universal.
Can IF/ICC help judge the compartment?Yes. HPA reports supported mitochondrial localization in U2OS ICC-IF images and lists CAB079899 as ICC Supported (HPA: subcellular location; HPA: antibody validation). That supports a mitochondrial IF comparison; it does not set a required punctate appearance for paraffin-section chromogenic IHC (HPA: general cytoplasmic IHC expression).
Why is my staining missing, weak or wrong?
SituationLikely causeNext action
Kidney tubules show no color.The result conflicts with their reported High staining; a failed staining or detection step remains possible (HPA: High in kidney tubule cells; standard IHC practice).Check the IHC control, reagent sequence and detection system, then review section quality before interpreting the sample as negative (standard IHC practice).
Only nuclei are colored.Nuclear-only signal does not fit the reported cytoplasmic and mitochondrial locations (HPA: tissue IHC; UniProt O43521: subcellular location).Compare with the counterstain and a negative control; reassess antibody specificity if the nuclear signal persists (standard IHC practice).
Smooth muscle or fibroblasts dominate the positive area.These populations are listed as Low in the supplied HPA examples; cross-reactivity or endogenous activity may contribute (HPA: tissue IHC; standard IHC interpretation).Compare the staining with a matched negative control and with an HPA High cell population in the same staining run (HPA: tissue IHC; standard IHC practice).
Brown haze obscures cell boundaries.Diffuse background prevents a reliable call of the cytoplasmic pattern (HPA: tissue IHC; standard IHC interpretation).Inspect the negative control and review blocking, washing and detection conditions as general chromogenic IHC checks (standard IHC practice).
Macrophages stain strongly while nearby cells are weak.This can reflect the reported cell-specific distribution: lung macrophages are High, while the HPA profile does not assign every nearby cell the same level (HPA: lung tissue IHC).Identify the stained cell population on morphology and score its cytoplasmic signal separately from adjacent cells (standard IHC practice).
IHC looks broadly cytoplasmic, but an IF image looks mitochondrial.Both observations can fit the supplied evidence: tissue IHC is described as generally cytoplasmic, and ICC-IF localization is supported at mitochondria (HPA: tissue IHC; HPA: subcellular location).Interpret each image at its own resolution; use the tissue IHC cell distribution for chromogenic scoring (HPA: tissue IHC; standard IHC interpretation).

Sample controls for BCL2L11 IHC & IF

🧪Run lung first and score macrophages for staining (HPA: High in lung macrophages). HPA detects BCL2L11 in all 45 scored tissues, so use no-primary and isotype controls instead of a negative tissue (HPA: no negative tissue rows); on the positive slide, treat any cells without specific staining as internal background comparators, without assuming a particular cell type is negative.
Positive control tissue: Appendix (Lymphoid tissue, HPA High)
Negative control tissue: None in HPA: BCL2L11 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 ICC-IF images show BCL2L11 in U2OS, with annotated localisation: Mitochondria (supported) (HPA subcellular).
Technical controls: Include a no-primary, secondary-only section, a rabbit immunoglobulin isotype control matched to the primary antibody’s clonality, and BCL2L11-knockout material as a biological specificity control (caption: rabbit primary; standard IHC control practice). Check lung macrophages for endogenous peroxidase activity and pigment that could mimic chromogenic signal (HPA: lung macrophages; standard IHC practice).
⚠️Feasibility: No target-specific fixation window or fixation effect is reported in the supplied evidence; the selected A01552-4 paraffin-section caption does not state a fixative (selected-SKU tissue-IHC caption). That caption reports heat-mediated retrieval in EDTA at pH 8.0, but does not establish whether retrieval is required for all specimens (selected-SKU tissue-IHC caption). IF images support mitochondrial localization in U2OS, but the supplied evidence does not establish whether frozen sections or IF are easier than paraffin IHC; assess lung macrophage pigment when interpreting chromogenic staining (HPA: subcellular ICC-IF; HPA: lung macrophages; standard IHC practice).

HPA tissue IHC evidence for BCL2L11

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

TissueCell typeLevelEvidenceSource
Appendix Lymphoid tissue High Protein (IHC) HPA →
Breast Adipocytes High Protein (IHC) HPA →
Endometrium Glandular cells High Protein (IHC) HPA →
Kidney Cells in tubules High Protein (IHC) HPA →
Lung Macrophages High Protein (IHC) HPA →

Undetected expression · recommended negative controls

TissueCell typeLevelEvidenceSource
None in HPA: BCL2L11 is detected in all 45 scored tissues. Use a no-primary (secondary-only) and an isotype control instead.
Section 3

Advanced BCL2L11 IHC Tips

Troubleshoot BCL2L11 staining in paraffin sections by checking retrieval, cellular context and controls before interpreting chromogenic signal (datasheet A01552-4; HPA tissue IHC).

Which retrieval conditions should I try when BCL2L11 staining is weak?
Start with heat-mediated retrieval in EDTA at pH 8.0 (datasheet A01552-4). The selected paraffin-section image used this buffer, followed by 10% goat serum blocking and 2 μg/ml primary antibody overnight at 4°C (datasheet A01552-4). If staining remains weak, compare retrieval duration on matched sections before considering another buffer or pH, and retain a no-primary control to identify retrieval-related background (standard IHC practice). Score cytoplasmic signal in the same cell type across conditions; HPA reports general cytoplasmic expression, while BCL2L11 is associated with intracytoplasmic membranes (HPA tissue IHC; UniProt O43521 subcellular location).
How should I troubleshoot fixation-related loss of BCL2L11 staining?
The selected tissue image identifies a paraffin section but does not report its fixative, so target-specific fixation sensitivity is unknown (datasheet A01552-4). Record the fixative, fixation duration and section age for each specimen, then compare matched sections using the same EDTA pH 8.0 retrieval and detection settings (datasheet A01552-4; standard IHC practice). Uneven penetration or prolonged processing can affect antigen accessibility in paraffin sections, so inspect morphology and staining across the section before changing antibody concentration (standard IHC practice). Include a concurrently processed positive-control section and a no-primary control; neither a staining pattern nor BCL2L11 phosphorylation establishes fixation tolerance (standard IHC practice; UniProt O43521 modified residues).
What BCL2L11 staining pattern is credible in paraffin sections?
Expect predominantly cytoplasmic staining in tissue sections, with possible membrane-associated intracellular detail (HPA tissue IHC; UniProt O43521 subcellular location). Supported mitochondrial localisation in cell imaging helps assess plausibility, but chromogenic tissue IHC alone does not resolve individual mitochondria reliably (HPA subcellular; standard IHC practice). BCL2L11 has no annotated transmembrane segment and is associated with intracytoplasmic membranes, so a sharp cell-surface rim or isolated nuclear signal needs independent validation (UniProt O43521 topology and subcellular location). Compare the suspect pattern with adjacent morphology, a no-primary control and a known positive compartment, keeping exposure and DAB development comparable between sections (standard IHC practice).
Can this antibody distinguish BimEL, BimL and BimS in IHC?
Do not assign a stained cell to one BCL2L11 isoform without an epitope map or isoform-specific validation for this antibody (standard IHC practice). BimEL, BimL and BimS are predominant, widely expressed isoforms, and the record lists 20 isoforms overall (UniProt O43521 isoforms and tissue specificity). Phosphorylation is annotated at several residues, including serine 69 and 77; the supplied tissue caption does not establish whether those changes affect antibody binding (UniProt O43521 modified residues; datasheet A01552-4). If isoform identity matters, pair cell-level IHC scoring with an independently validated isoform-specific assay and report the IHC result as BCL2L11 immunoreactivity (standard IHC practice).
How can IF clarify ambiguous BCL2L11 signal seen by chromogenic IHC?
Use IF as a follow-up localisation check rather than interpreting fine mitochondrial detail from DAB alone (HPA subcellular; standard IHC practice). In lung sections, multiplex BCL2L11 with a validated macrophage marker such as CD68, since HPA reports high BCL2L11 staining in lung macrophages (HPA tissue IHC; standard IF practice). Choose spectrally separated fluorophores and assess unstained tissue autofluorescence, favoring a longer-wavelength channel for the weaker signal when background permits (standard IF practice). Because BCL2L11 is associated with intracytoplasmic membranes but its antibody epitope and membrane-facing side are unspecified, optimise gentle permeabilisation empirically and compare it with an unpermeabilised control (UniProt O43521 subcellular location; standard IF practice).
How do I reduce diffuse or punctate background in BCL2L11 IHC?
First compare a no-primary section with the stained section to separate detection-system deposits from antibody-dependent signal (standard IHC practice). The selected image used 10% goat serum, a biotinylated secondary antibody, a streptavidin–biotin complex and DAB, so assess endogenous peroxidase and endogenous biotin background in that workflow (datasheet A01552-4; standard IHC practice). Keep blocking, washes and DAB development consistent while adjusting one variable at a time; inspect folds, edges and necrotic areas separately from intact cells (standard IHC practice). Judge residual signal against cytoplasmic staining in expected cells, including lung macrophages, and avoid treating every brown granule as BCL2L11 (HPA tissue IHC; standard IHC practice).
How should I quantify BCL2L11 across heterogeneous tissue sections? ⚠ ANSWER MARKED FOR VERIFICATION
Define the cell population and region before scoring, then record the percentage of positive cells and staining intensity within that population (standard IHC practice). An H-score combines intensity categories with their percentages on a 0–300 scale; alternatively report positive-cell density per mm² when cell abundance is the primary question (standard IHC practice). Normalise counts to the number of eligible cells or sampled tissue area, and keep section thickness, retrieval, imaging and DAB development comparable (standard IHC practice). Separate macrophages from other lung cells where relevant, since HPA reports high staining in lung macrophages and medium consistency between antibody staining and RNA expression (HPA tissue IHC).
When is a BCL2L11-positive cell convincing rather than an artefact?
A convincing result is cellular cytoplasmic staining in intact tissue, interpreted alongside morphology and a no-primary control (HPA tissue IHC; standard IHC practice). Lung macrophages provide a documented positive cell context, whereas a signal restricted to necrosis, tissue edges or folds is less persuasive (HPA: high in lung macrophages; standard IHC practice). Diffuse nuclear staining or a crisp plasma-membrane outline warrants review because BCL2L11 is linked to mitochondria and intracytoplasmic membranes (HPA subcellular; UniProt O43521 subcellular location). Check endogenous peroxidase and biotin contributions when using the selected biotin–streptavidin DAB workflow, and confirm disputed cell identities with morphology or an independent marker (datasheet A01552-4; standard IHC practice).
Boster reagents

Best BCL2L11 / Bcl-2-like protein 11 IHC Antibodies

The catalog shows human paraffin-section IHC examples (A01552-4, M01552, A01552-3: image captions) and K562 IF/ICC examples (A01552, A01552-2: image captions).

Real IHC data IHC analysis of Bim/BCL2L11 using anti-Bim/BCL2L11 antibody (A01552-4). Bim/BCL2L11 was detected in a paraffin-embedded section of human lung cancer 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-Bim/BCL2L11 Antibody (A01552-4) 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.
Anti-Bim/BCL2L11 Antibody ®
Cat # A01552-4
Real IHC data Immunohistochemical analysis of paraffin-embedded Human prostate cancer, using the Antibody at 1:50 dilution.
Anti-Bim BCL2L11 Rabbit Monoclonal Antibody
Cat # M01552
Real IHC data Formalin-fixed and paraffin-embedded human colon carcinoma reacted with BCL2L11 Antibody (Center), which was peroxidase-conjugated to the secondary antibody, followed by DAB staining. This data demonstrates the use of this antibody for immunohistochemistry; clinical relevance has not been evaluated.
Anti-BCL2L11 Antibody (Center)
Cat # A01552-3
Real IHC data Induced Expression Validation of BIM in Mouse Hippocampus (Tsuchiya et al., 2011) The induction of Bim protein was detected by immunohistochemical analysis of mice after i.h. injection of epoxomicin with anti-BIM antibodies. Sections from epoxomicin-treated animals exhibited cells staining positive for Bim expression within the NeuN-positive population of neurons in the CA1 of the ipsilateral side. In contrast, Bim-positive cells were absent within the NeuNpositive CA1 neurons on the contralateral side.
Anti-Bim BCL2L11 Antibody
Cat # A01552
Real IF data Immunocytochemistry of BIM EL inK562 cells with BIM EL antibody at 2.5 μg/ml.
Anti-Bim EL BCL2L11 Antibody
Cat # A01552-2

A01552-4 shows human lung cancer IHC (catalog image caption); M01552 shows human prostate cancer IHC (catalog image caption); and A01552-3 shows formalin-fixed, paraffin-embedded human colon carcinoma IHC (catalog image caption). A01552 shows IF in K562 cells and IHC in mouse hippocampus (catalog image captions); A01552-2 shows ICC/IF in K562 cells (catalog image captions).

Which to pick: For human paraffin-section IHC, start with A01552-4 at 2–5 μg/ml (catalog: IHC dilution); its lung cancer caption specifies EDTA retrieval at pH 8.0 but does not report the fixative (A01552-4: IHC caption). For IF/ICC, choose A01552 for human samples (catalog: application validation) or A01552-2 when targeting the Bim EL isoform (catalog: title and IF/ICC captions). For a monoclonal option with listed human, mouse and rat reactivity across IHC and IF/ICC, choose M01552 (catalog: clone CBE-2, applications and reactivity); its paraffin-section IHC caption does not report the fixative (M01552: IHC caption).

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

References

  1. UniProt Consortium. UniProt entry O43521 (B2L11_HUMAN, Bcl-2-like protein 11).
  2. Human Protein Atlas. BCL2L11 tissue IHC expression (reliability: Approved).
  3. Human Protein Atlas. BCL2L11 subcellular location (ICC-IF): Localized to the mitochondria..
  4. Human Protein Atlas. BCL2L11 antibody validation summary (2 antibodies).
  5. Plasma microRNA panel is a novel biomarker for focal segmental glomerulosclerosis and associated with podocyte apoptosis. Cell death & disease 2018 — PMC5945632.
  6. Identification of BCL2L11 as a Candidate Gene for Hereditary Predisposition to Non-Medullary Thyroid Cancer Using Familial Whole-Exome-Sequencing. Clinical genetics 2026 — PMC12881209.
  7. Machine learning deciphers the significance of mitochondrial regulators on the diagnosis and subtype classification in non-alcoholic fatty liver disease. Heliyon 2024 — PMC11066337.
  8. A urokinase receptor-Bim signaling axis emerges during EGFR inhibitor resistance in mutant EGFR glioblastoma. Cancer research 2015 — PMC4297573.
  9. PubMed PMID:9430630 — UniProt-cited evidence.
  10. PubMed PMID:11734221 — UniProt-cited evidence.
  11. PubMed PMID:12019181 — UniProt-cited evidence.