This website uses cookies to ensure you get the best experience on our website.
- Table of Contents
BCL2L1 / Bcl-2-like protein 1 · IHC design guide
Design Immunohistochemistry for BCL2L1
Plan chromogenic BCL2L1 IHC around the cytoplasmic staining reported in select tissues (HPA tissue IHC). This guide covers fixation consistency, tissue controls and antibody epitope coverage across 3 isoforms (standard IHC practice; UniProt).
Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
BCL2L1 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 tissue staining (HPA tissue IHC) | |
| Staining pattern | Cytoplasmic signal in marrow, lymph node and tubule cells (HPA tissue IHC) | |
| Antigen retrieval | EDTA pH 8.0 HIER, heat-mediated (datasheet PA1232) | |
| 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 paraffin sections (standard IHC practice; not target-specific) | |
| Caveat | Antibody staining and RNA show low consistency (HPA tissue IHC) | |
| Regulation | Bcl-X(S) high in developing lymphocytes (UniProt) | |
| Isoform / epitope | 3 splice isoforms; confirm antibody epitope coverage (UniProt) |
Section 1
Recommended BCL2L1 IHC & IF Protocols
The catalog antibody’s IHC-P protocol (datasheet: PA1232) is accompanied by one published colorectal tumor IHC protocol (PMC4456718).
| Sample | Tissue sections; selected-image fixative not specified (standard IHC workflow) |
| Fixation | Image fixative and duration unreported (datasheet PA1232); 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 PA1232); 20 min, 95–100 °C (standard) |
| Peroxidase block | 3% H2O2, 10 min, room temperature (standard) |
| Blocking | 10% goat serum (datasheet PA1232) |
| Primary antibody | Rabbit anti-BCL2L1, 0.5-1μg/ml (datasheet PA1232) |
| Primary incubation | Overnight at 4 °C (datasheet PA1232) |
| Detection | HRP-conjugated secondary, DAB chromogen (datasheet PA1232) |
| Counterstain | Hematoxylin, blue, dehydrate and mount (standard) |
| Expected result | BCL2L1-positive staining in glandular cells of adrenal gland (HPA tissue IHC: Medium). HPA tissue profile: Cytoplasmic expression in a few tissues, including bone marrow, lymph node and cells in tubules. No signal in the no-primary control. |
💡Decision noteStart with heat-mediated EDTA retrieval at pH 8.0 for the catalog antibody (datasheet: PA1232); the published study does not specify retrieval (PMC4456718: Methods).
Section 2
What Is the Expected BCL2L1 Staining Pattern?
BCL2L1 is a membrane associated protein with a transmembrane segment at residues 210–226 and reported mitochondrial, cytosolic, vesicular, and other locations (UniProt Q07817 topology and subcellular location). In paraffin section IHC, expect mainly cytoplasmic staining in selected cells, including bone marrow hematopoietic cells, lymph node non-germinal center cells, and kidney tubular cells (HPA tissue IHC). HPA rates the tissue staining Approved but reports low consistency with RNA expression (HPA tissue IHC).
| Moderate cytoplasmic staining in kidney tubular cells, bone marrow hematopoietic cells, or lymph node non-germinal center cells (HPA tissue IHC). | This matches the reported positive cell populations and compartment (HPA tissue IHC). Score the named cells rather than calling the entire section positive. HPA reports low consistency between antibody staining and RNA expression, so this pattern supports interpretation without independently proving antibody specificity (HPA tissue IHC). |
| Predominantly nuclear chromogen, with little cytoplasmic staining in the expected cells. | Treat a nuclear dominant IHC pattern as questionable: HPA describes tissue staining as cytoplasmic, while UniProt lists a nuclear membrane location among several locations (HPA tissue IHC; UniProt Q07817 subcellular location). Check the counterstain and detection controls before assigning that pattern to BCL2L1 (general IHC practice). |
| Strong staining in adipocytes or heart cardiomyocytes, with weak staining in the expected positive cells. | HPA reports BCL2L1 as not detected in those two cell populations (HPA tissue IHC). Consider cross-reactivity or endogenous detection activity, especially if a no-primary control also stains (general IHC practice). A single unexpected positive field does not establish a new tissue pattern. |
| Diffuse chromogen across stroma, cell borders, and empty areas rather than within identifiable cells. | This distribution does not resemble HPA's cell-restricted cytoplasmic tissue pattern (HPA tissue IHC). Review the no-primary control for detection background and assess washing and blocking (general IHC practice). Interpret intensity only after background allows individual positive cells to be distinguished. |
| No staining in kidney tubular cells or bone marrow hematopoietic cells. | Both are reported as Medium positive by HPA, so absence of signal calls for a run-level check (HPA tissue IHC). Confirm that a known-positive control worked, then review section handling, antigen retrieval, antibody application, and chromogen detection (general IHC practice). HPA's low RNA concordance limits how much one negative specimen can prove (HPA tissue IHC). |
💡Expected BCL2L1 appearanceCall a result positive when identifiable kidney tubular cells, bone marrow hematopoietic cells, or lymph node non-germinal center cells show Medium cytoplasmic chromogen (HPA tissue IHC); diffuse background or dominant nuclear staining is a questionable pattern against that reference (HPA tissue IHC; general IHC practice).
| Cell population chosen for scoring (HPA tissue IHC). | HPA reports Medium staining in selected cells across several tissues, including respiratory epithelial cells in bronchus and glandular cells in adrenal gland (HPA tissue IHC). It also reports Not detected in specific cells within other tissues; compare like cell populations rather than whole organs (HPA tissue IHC). |
| Tissue IHC evidence and antibody validation (HPA tissue IHC; HPA antibodies). | The HPA tissue assessment is Approved but notes low agreement between staining and RNA data; HPA035734 has Approved IHC status (HPA tissue IHC; HPA antibodies). Use the reported staining as a reference pattern, with a positive and detection control for the run (general IHC practice). |
| Membrane association and cellular compartment (UniProt Q07817 topology and subcellular location). | A transmembrane segment at residues 210–226 accompanies UniProt mitochondrial membrane, synaptic vesicle membrane, cytosolic, and other location annotations (UniProt Q07817). These locations inform interpretation, but HPA's paraffin section reference is cytoplasmic staining (HPA tissue IHC). |
| Isoforms and tissue context (UniProt Q07817 isoforms and tissue specificity). | UniProt lists Bcl-X(L), Bcl-X(S), and Bcl-X(beta); it describes Bcl-X(S) in high-turnover cells such as developing lymphocytes and Bcl-X(L) in long-lived cells such as adult brain (UniProt Q07817). The supplied record gives no antibody epitope or isoform selectivity, so IHC intensity cannot identify an isoform. |
| IF/ICC Q: Should punctate mitochondrial fluorescence be expected? | A: HPA calls mitochondria the approved main ICC-IF location; nucleoli fibrillar center is uncertain, and acrosome is approved (HPA subcellular). That IF observation does not set a required punctate appearance for chromogenic paraffin section IHC, whose HPA tissue description is cytoplasmic (HPA tissue IHC). |
| Situation | Likely cause | Next action |
|---|---|---|
| Expected positive cells are blank while the no-primary control is clean. | The run may have failed at section preparation, retrieval, primary antibody application, or detection (general IHC practice); HPA reports Medium staining in kidney tubular cells and bone marrow hematopoietic cells (HPA tissue IHC). | Check a known-positive section processed in the same run, then review each IHC step and reagent record (general IHC practice). Avoid calling the specimen biologically negative until the control produces interpretable staining. |
| Chromogen appears in the no-primary control. | Detection-system or endogenous activity can produce antibody-independent signal (general IHC practice). Such signal cannot be counted as BCL2L1 even if it overlaps a cell population reported positive by HPA (HPA tissue IHC). | Review endogenous enzyme blocking, secondary detection, and chromogen controls; resolve the control signal before scoring the sample (general IHC practice). |
| Most cells and extracellular areas show similar color. | High nonspecific background can obscure HPA's selective cytoplasmic pattern (HPA tissue IHC; general IHC practice). | Compare with the no-primary control, then review blocking, washes, and antibody concentration according to the antibody's validated IHC instructions (general IHC practice). Score only after cell boundaries and the expected positive populations are distinguishable. |
| Staining is nuclear dominant in every tissue compartment. | The distribution conflicts with HPA's cytoplasmic tissue description; a listed nuclear membrane location does not establish diffuse nuclear IHC staining (HPA tissue IHC; UniProt Q07817 subcellular location). | Check counterstain interpretation and no-primary staining, then compare a validated positive section in the same run (general IHC practice). |
| Adipocytes or cardiomyocytes stain more strongly than the expected positive cells. | HPA reports those populations as Not detected, making cross-reactivity or detection background plausible explanations (HPA tissue IHC; general IHC practice). | Confirm cell identity on the section, inspect the no-primary control, and compare with an HPA-reported positive population before attributing the unexpected signal to BCL2L1 (HPA tissue IHC; general IHC practice). |
| An IF image looks mitochondrial, but the matched IHC section looks broadly cytoplasmic. | HPA assigns mitochondria as the approved main ICC-IF location and describes tissue IHC staining as cytoplasmic (HPA subcellular; HPA tissue IHC). The two readouts have different spatial resolution (general microscopy practice). | Judge each image against its own HPA reference and controls. Use the tissue IHC cell population and cytoplasmic distribution to score the paraffin section (HPA tissue IHC; general IHC practice). |
Experiment evidence
IHC & IF images showing BCL2L1 staining
PA1232 · IHCAnti-Bcl-XS antibody, PA1232, IHC(P) IHC(P): Human Colon Cancer Tissue
PA1232 · IHCAnti-Bcl-XS antibody, PA1232, IHC(P) IHC(P): Human Colon Cancer Tissue
A00181 · IHCIHC analysis of Bcl-XL using anti-Bcl-XL antibody (A00181). Bcl-XL was detected in a paraffin-embedded section of human pancrease cancer tis
A00181 · IFIF analysis of Bcl-XL using anti-Bcl-XL antibody (A00181). Bcl-XL was detected in an immunocytochemical section of TPC-1 cells. Enzyme antig
M00181-1 · IHCImmunohistochemical analysis of paraffin-embedded Human pituitary tumor, using the Antibody at 1:400 dilution.
M00181-1 · IHCImmunohistochemical analysis of paraffin-embedded Human liver cancer, using the Antibody at 1:400 dilution.
M00181-1 · IFImmunofluorescent analysis using the Antibody at 1:50 dilution.
M00181-4 · IHCHuman colon carcinoma was stained with anti-Bcl-XL rabbit antibody
M00181-4 · IHCHuman colon carcinoma was stained with anti-Bcl-XL rabbit antibody
HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000171552-BCL2L1/tissue/cerebral+cortex#imid_17702407
HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000171552-BCL2L1/tissue/colon#imid_17702333
HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000171552-BCL2L1/tissue/liver#imid_17702342
HPA · KidneyHuman Protein Atlas IHC, Kidney. Source: https://www.proteinatlas.org/ENSG00000171552-BCL2L1/tissue/kidney#imid_17702335
HPA · ICC-IF A-431Human Protein Atlas immunofluorescence, A-431, antibody HPA035734; green = BCL2L1, blue = nucleus, red = microtubules.
HPA · ICC-IF A-431Human Protein Atlas immunofluorescence, A-431, antibody HPA035734; green = BCL2L1, blue = nucleus, red = microtubules.
← swipe / scroll for more images →
Real IHC / IF data behind the staining-pattern expectations in this section.
Sample controls for BCL2L1 IHC & IF
🧪Run bronchus first: respiratory epithelial cells should show medium staining (HPA: Bronchus, respiratory epithelial cells, Medium). Use nasopharynx respiratory epithelium as the negative comparison (HPA: Nasopharynx, respiratory epithelial cells, Not detected); unstained cells elsewhere on the bronchus slide can indicate background, but the supplied HPA rows do not establish a specific internal-negative cell type.
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 BCL2L1 in A-431, A-549, U2OS, HeLa, MCF-7, Sperm, with annotated localisation: Mitochondria (approved) (HPA subcellular).
Technical controls: Include a no-primary, secondary-only control; an isotype control matched to the primary antibody’s host species and clonality; and, if available, a BCL2L1 knockout sample or immunizing-peptide block (standard IHC controls). For chromogenic IHC, quench endogenous peroxidase and check endogenous biotin if using avidin–biotin detection; for IF, assess autofluorescence with an unstained section (standard IHC/IF practice).
⚠️Feasibility: A target-specific fixation window and fixation effect are unreported in the supplied evidence; the PA1232 paraffin-section caption does not report a fixative (caption: IHC(P), fixative not stated). Antigen retrieval dependence is also unreported, so optimize retrieval for paraffin sections against the controls (standard IHC practice). The evidence does not establish that frozen sections or IF are easier, and it reports no bronchus-specific artefact; ICC-IF images support mitochondrial localization but do not establish frozen-section performance (HPA subcellular: Mitochondria approved).
HPA tissue IHC evidence for BCL2L1
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 |
|---|---|---|---|---|
| Adrenal gland | Glandular cells | Medium | Protein (IHC) | HPA → |
| Bone marrow | Hematopoietic cells | Medium | Protein (IHC) | HPA → |
| Bronchus | Respiratory epithelial cells | Medium | Protein (IHC) | HPA → |
| Cerebral cortex | Endothelial cells | Medium | Protein (IHC) | HPA → |
| Fallopian tube | Glandular cells | Medium | Protein (IHC) | HPA → |
Undetected expression · recommended negative controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| Adipose tissue | Adipocytes | Not detected | Protein (IHC) | HPA → |
| Cervix | Glandular cells | Not detected | Protein (IHC) | HPA → |
| Heart muscle | Cardiomyocytes | Not detected | Protein (IHC) | HPA → |
| Hippocampus | Glial cells | Not detected | Protein (IHC) | HPA → |
| Nasopharynx | Respiratory epithelial cells | Not detected | Protein (IHC) | HPA → |
Section 3
Advanced BCL2L1 IHC Tips
Troubleshoot BCL2L1 staining in paraffin sections by checking retrieval, cell identity, subcellular pattern and controls before comparing signal across samples.
What retrieval should I try first when BCL2L1 staining is weak in paraffin sections?
Start with heat-mediated retrieval in EDTA at pH 8.0 for PA1232 (datasheet PA1232). Run a short heating-time series, such as 10–20 minutes, while keeping section thickness, antibody incubation and detection constant (standard IHC practice). Compare staining in kidney tubular cells with a no-primary control, because these cells show medium tissue staining in the reference profile (HPA: kidney cells in tubules, Medium). If staining remains weak, test citrate at pH 6.0 as a fallback on adjacent sections (standard IHC practice). Select the condition that preserves morphology and gives cellular signal without increased background (standard IHC practice).
Could fixation explain weak BCL2L1 staining in my paraffin sections?
Target-specific fixation sensitivity is unknown from the supplied evidence; the PA1232 tissue caption reports paraffin IHC in human colon cancer tissue but does not state a fixative (PA1232 tissue-IHC caption). Document the fixative and fixation interval for every specimen before comparing staining intensity (standard IHC practice). If processing varies, stain adjacent sections together and compare them with a consistently processed reference section and a no-primary control (standard IHC practice). Avoid assigning a weak result to fixation alone until retrieval, tissue preservation and detection have been checked (standard IHC practice). Treat any fixation-time comparison as a local validation, not an established BCL2L1 requirement.
Where should convincing BCL2L1 staining appear within positive cells?
Expect predominantly intracellular staining with a mitochondrial pattern, which is the approved principal subcellular location (HPA subcellular: mitochondria, approved). Cytosolic, vesicular, centrosomal and nuclear-membrane localisations are also annotated, so diffuse cytoplasmic signal is not automatically implausible (UniProt Q07817 subcellular). The protein has a transmembrane segment at residues 210–226, supporting membrane association but not proving that a chromogenic punctum identifies a mitochondrion (UniProt Q07817 topology). Compare the pattern with cell morphology and, where needed, a separately validated mitochondrial marker on adjacent sections (standard IHC practice). Reassess staining confined to tissue edges or damaged regions before calling a new compartment (standard IHC practice).
Can PA1232 staining distinguish Bcl-X(S) from other BCL2L1 isoforms?
PA1232 is captioned as an anti-Bcl-XS antibody, but the supplied evidence does not map its epitope or establish isoform-exclusive staining in sections (PA1232 tissue-IHC caption). BCL2L1 has 3 annotated isoforms: Bcl-X(L), Bcl-X(S) and Bcl-X(beta) (UniProt Q07817 isoforms). Before making an isoform-specific claim, obtain the immunogen or epitope sequence and check whether it is unique to the intended splice form (standard IHC validation practice). Include an independently validated isoform-specific assay if the distinction determines the conclusion (standard IHC validation practice). Report tissue staining as antibody-reactive signal while that specificity remains unresolved.
How should I investigate BCL2L1 localisation by IF alongside this IHC guide?
Use the separate IF/ICC workflow for fluorescence localisation; HPA reports mitochondria as the approved main location in its subcellular images (HPA subcellular: mitochondria, approved). Multiplex with a validated marker for the expected cell population, such as kidney tubular cells, and check that signals occur in the same cells (HPA tissue: kidney cells in tubules, Medium; standard IF practice). Choose a red or far-red fluorophore after measuring tissue autofluorescence and include single-stain controls (standard IF practice). The 210–226 transmembrane segment does not identify which side contains the antibody epitope (UniProt Q07817 topology). Determine epitope accessibility before selecting permeabilisation strength, then compare mild and stronger conditions without assuming either exposes this antibody’s target (standard IF practice).
How can I reduce diffuse or granular background without losing real signal?
First compare the stained section with a no-primary section to identify detection-related colour, then inspect an adjacent stained section for cellular pattern (standard IHC practice). Block endogenous peroxidase before chromogenic detection, use an appropriate protein block, and titrate primary antibody and incubation time against background (standard IHC practice). Keep washes and DAB development consistent across the series so a darker slide is not mistaken for higher expression (standard IHC practice). Because mitochondria are an approved main location, fine intracellular granularity can be plausible, but staining spread across empty spaces or section edges needs scrutiny (HPA subcellular: mitochondria, approved; standard IHC practice). Record which adjustment improves cell-associated contrast.
How should I score BCL2L1 IHC across samples with different cell composition? ⚠ ANSWER MARKED FOR VERIFICATION
Define the cell population and region before scoring; the reference profile reports medium staining in kidney tubular cells and lymph-node non-germinal-center cells (HPA tissue: kidney and lymph node, Medium). For cytoplasmic chromogenic staining, record the percentage of positive target cells and intensity, then calculate an H-score from 0–300 if intensity categories are reproducible (standard IHC scoring practice). Alternatively, report positive-cell density per mm² when cell abundance is the outcome (standard IHC scoring practice). Normalise to the number of eligible cells or analysed tissue area, exclude damaged and necrotic regions, and keep thresholds fixed across batches (standard IHC practice). Report cell-type proportions separately from staining intensity.
When should I doubt an apparent BCL2L1-positive result?
Give greatest weight to cell-associated cytoplasmic or mitochondrial-pattern staining in an anatomically identified population, consistent with the reference localisation (HPA subcellular: mitochondria, approved). Check whether the purported positive cells match the population being scored; the tissue profile includes positive kidney tubular cells but also several cell types reported as not detected (HPA tissue profile). Edge-restricted colour, necrotic areas and signal in the no-primary control suggest section or detection artefact (standard IHC practice). Repeat the peroxidase block or review DAB development if endogenous enzyme activity is suspected (standard IHC practice). Interpret a discordant result cautiously because the tissue-IHC profile has low consistency with RNA expression (HPA tissue: reliability description).
Boster reagents
Best BCL2L1 / Bcl-2-like protein 1 IHC Antibodies
IHC images show human tissues (catalog IHC captions: PA1232, A00181, M00181-1, M00181-4); IF images show TPC-1 and HeLa cells (catalog IF captions: A00181, M00181-1). All four list human, mouse and rat reactivity (catalog reactivity).
Real IHC data
Anti-Bcl-XS/BCL2L1 Antibody ®
Cat # PA1232
Real IHC data
Anti-Bcl-XL/BCL2L1 Antibody ®
Cat # A00181
Real IHC data
Anti-Bcl-XL BCL2L1 Rabbit Monoclonal Antibody
Cat # M00181-1
Real IHC data
Anti-Bcl-XL Rabbit Monoclonal Antibody
Cat # M00181-4
PA1232 shows paraffin-section IHC in human colon cancer, while A00181 shows paraffin-section IHC in human pancreatic cancer and IF in TPC-1 cells (PA1232 and A00181 image captions). M00181-1 shows paraffin-section IHC in human pituitary tumor and liver cancer plus IF in HeLa cells; M00181-4 shows IHC in human colon carcinoma (M00181-1 and M00181-4 image captions).
Which to pick: For tissue IHC targeting Bcl-XL, choose A00181: its paraffin-section caption documents EDTA retrieval at pH 8.0 and primary antibody at 2 μg/ml; the fixative is unreported (A00181 IHC caption). For IF/ICC, A00181 has a TPC-1 cell image at 5 μg/ml (A00181 IF caption). For mouse or rat IHC planning, M00181-4 is a monoclonal option with both species listed as reactive, although its IHC image shows human colon carcinoma (M00181-4 catalog description, reactivity and IHC caption).
Each figure is that product's own IHC / IF validation image from its datasheet.
References
- UniProt Consortium. UniProt entry Q07817 (B2CL1_HUMAN, Bcl-2-like protein 1).
- Human Protein Atlas. BCL2L1 tissue IHC expression (reliability: Approved).
- Human Protein Atlas. BCL2L1 subcellular location (ICC-IF): Mainly localized to the mitochondria. In addition localized to the nucleoli fibrillar center and acrosome..
- Human Protein Atlas. BCL2L1 antibody validation summary (2 antibodies).
- BuFeiXiaoJiYin ameliorates the NLRP3 inflammation response and gut microbiota in mice with lung cancer companied with Qi-yin deficiency. Cancer cell international 2022 — PMC8922837.
- Gene expression profiling of laterally spreading tumors. BMC gastroenterology 2015 — PMC4456718.
- Expression and gene regulation network of TYMS and BCL2L1 in colorectal cancer based on data mining. PeerJ 2021 — PMC8179227.
- Molecular mechanism of the miR-7/BCL2L1/P53 signaling axis regulating the progression of hepatocellular carcinoma. Annals of translational medicine 2023 — PMC9906214.
- PubMed PMID:8358789 — UniProt-cited evidence.
- PubMed PMID:9675101 — UniProt-cited evidence.
- PubMed PMID:17974005 — UniProt-cited evidence.
Immunohistochemistry protocol sheet for BCL2L1 (Bcl-2-like protein 1). Expected localisation Cytoplasmic tissue staining (HPA tissue IHC). Antigen retrieval: EDTA pH 8.0 HIER, heat-mediated (datasheet PA1232). Validated IHC antibody: PA1232 at 0.5-1μg/ml (the product's own IHC validation image is shown). Controls (HPA): positive Adrenal gland (Glandular cells); negative Adipose tissue. IHC workflow; specimen preparation unspecified. Protocol steps: 1. Sample: Tissue sections; selected-image fixative not specified (standard IHC workflow); 2. Fixation: Image fixative and duration unreported (datasheet PA1232); verify before use.; 3. Sectioning: 4–5 µm sections on charged slides (standard); 4. Deparaffinisation: Xylene, graded ethanol series to water (standard); 5. Antigen retrieval: Heat retrieval: EDTA pH 8.0 (datasheet PA1232); 20 min, 95–100 °C (standard); 6. Peroxidase block: 3% H2O2, 10 min , room temperature (standard); 7. Blocking: 10% goat serum (datasheet PA1232); 8. Primary antibody: Rabbit anti-BCL2L1, 0.5-1μg/ml (datasheet PA1232); 9. Primary incubation: Overnight at 4 °C (datasheet PA1232). Watch for: Fixation: Keep fixation consistent across paraffin sections (standard IHC practice; not target-specific); Caveat: Antibody staining and RNA show low consistency (HPA tissue IHC); Isoform / epitope: 3 splice isoforms; confirm antibody epitope coverage (UniProt); Regulation: Bcl-X(S) high in developing lymphocytes (UniProt). Evidence: UniProt Q07817, HPA tissue IHC · Approved, HPA ICC-IF · Mainly localized to the mitochondria. In addition localized to the…, Datasheet PA1232, PMC4456718. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/BCL2L1/immunohistochemistry-ihc-guide.