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BCL2L2 / Bcl-2-like protein 2 · Western blot design guide
Design a Western Blot for BCL2L2
Real validated BCL2L2 Western blot protocols, expected-band and isoform facts, troubleshooting for weak or shifted signal, and recommended anti-BCL2L2 WB antibodies. Everything you need to plan the experiment before you commit precious samples.
Evidence assembled September 2026 · For research use; verify linked source records and product datasheet before use
BCL2L2 Western Blot Experimental Design Guide
Expected bands, validated protocols, controls and antibodies — the at-a-glance facts below, then the full design guide.
Must know before running
| Expected band | ~20.7 kDa | |
| Observed band | ~21 kDa | |
| Gel | 5–20% (catalog PA1426-1) | |
| Positive control | Cerebral cortex (IHC candidate; verify WB) +4 more | |
| Negative control | Oral mucosa (IHC candidate; verify WB) |
Important caveats
Reasons your observed band may differ from the expected size.
| PTM | Phosphorylated + Acetylated | |
| Caveat | Phosphorylation-state controls | |
| Gene-set association | MSigDB Hallmark membership | |
| Isoform | 2 isoform(s) |
Section 1
Real Curated BCL2L2 Western Blot Protocols
The PA1426-1 protocol combines labelled catalog values with standard starting conditions. Published comparisons retain their own sample, reagent and detection scope.
| Sample / lysate | human A549, human K562, human U-937 (catalog PA1426-1) |
| Gel % | 5–20% (catalog PA1426-1) |
| Load | 50ug; reducing conditions (catalog PA1426-1) |
| Transfer | a Nitrocellulose membrane at 150mA for 50-90 minutes (catalog PA1426-1) |
| Membrane | Nitrocellulose membrane (catalog PA1426-1) |
| Blocking | 5% Non-fat Milk/ TBS for 1.5 hour at RT (catalog PA1426-1) |
| Primary antibody | PA1426-1 · 0.5 μg/mL (catalog PA1426-1) |
| Primary incubation | overnight at 4°C (catalog PA1426-1) |
| Secondary antibody | goat anti-rabbit IgG-HRP, 1:10000 (catalog PA1426-1) |
| Secondary incubation | 1.5 hour at RT (catalog PA1426-1) |
| Wash | TBS-0.1%Tween 3 times with 5 minutes each (catalog PA1426-1) |
| Detection | ECL (catalog PA1426-1) |
Section 2
What Is the Expected BCL2L2 Western Blot Band Size?
BCL2L2 is predicted at 20.7 kDa and observed at ~21 kDa; the supplied features do not establish a cause for the small difference.
| Band at ~21 kDa | Observed BCL2L2 signal in reducing whole-cell lysates |
| Single sharp band near 21 kDa | Isoforms 1 and 3 need not produce separate visible bands |
| Additional bands at different positions | Isoforms 1 and 3 are possible contributors; their migration is not established |
| Faint band in a soluble fraction | BCL2L2 associates with the mitochondrial membrane |
💡Expected BCL2L2 appearanceBCL2L2 has a predicted mass of 20.7 kDa and an observed band at ~21 kDa in reducing whole-cell lysates; confirm band identity with appropriate controls.
| Predicted BCL2L2 mass | 20.7 kDa is the sequence-based reference; the observed band is ~21 kDa |
| Isoform 1 | Its individual apparent size is not supplied |
| Isoform 3 | Its individual apparent size is not supplied |
| Alternative splicing of isoforms 1 and 3 | Could affect band positions, but distinct migration is not established |
| Situation | Likely cause | Next action |
|---|---|---|
| No band in lysate | Membrane-associated BCL2L2 may be poorly extracted | Check extraction and a mitochondrial membrane fraction |
| Band higher than expected | The supplied features do not establish a size-increasing mechanism | Compare with the ~21 kDa reference and verify identity by BCL2L2 depletion |
| Band lower than expected | The supplied features do not establish a cleavage product | Verify identity by BCL2L2 depletion and check sample integrity |
| Multiple bands | Isoforms 1 and 3 are annotated, but separate migration is unproven | Use BCL2L2 depletion and isoform-specific evidence to identify bands |
| Weak or no signal | Recovery of mitochondrial membrane-associated BCL2L2 may be low | Check membrane protein recovery and sample loading |
Experiment evidence
Western blot images showing BCL2L2 band
PA1426-1 · BCL2L2
← swipe / scroll for more images →
Real WB data behind the band-size expectations in this section.
Sample controls for BCL2L2 Western blot
🧪HPA-IHC candidate guidance (verify in WB): For positive controls for BCL2L2 in Western blot, you can use cerebral cortex lysate, which HPA rates High.
Positive control: Cerebral cortex (IHC candidate; verify WB)
Negative control: Oral mucosa (IHC candidate; verify WB)
Loading controls: Run GAPDH, β-actin, and a total-protein stain such as Ponceau alongside the samples.
⚠️Feasibility: BCL2L2 is loosely associated with mitochondrial membranes in healthy cells, so consistent membrane recovery matters.
HPA tissue expression evidence for BCL2L2
Comprehensive Human Protein Atlas IHC scoring per tissue. Rows are taken directly from the HPA tissue chart — click any row's HPA link to view the source.
Higher expression tissues · candidate positive controls from IHC
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| Cerebral cortex | endothelial cells | High | Protein (IHC) | HPA → |
| Endometrium | glandular cells | High | Protein (IHC) | HPA → |
| Gallbladder | glandular cells | High | Protein (IHC) | HPA → |
| Lung | macrophages | High | Protein (IHC) | HPA → |
| Skeletal muscle | myocytes | High | Protein (IHC) | HPA → |
Lower expression tissues · IHC evidence, not confirmed WB-negative controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| Oral mucosa | squamous epithelial cells | Not detected | Protein (IHC) | HPA → |
| Ovary | ovarian stroma cells | Not detected | Protein (IHC) | HPA → |
| Adipose tissue | adipocytes | Medium | Protein (IHC) | HPA → |
| Adrenal gland | glandular cells | Medium | Protein (IHC) | HPA → |
| Appendix | glandular cells | Medium | Protein (IHC) | HPA → |
Section 3
Advanced BCL2L2 Western Blot Tips
Deeper troubleshooting and optimisation questions for BCL2L2, answered from its protein features.
How should BCL2L2 band migration be interpreted?
Band shift · Use the separately labelled calculated mass and catalog-observed evidence above. A sequence annotation does not establish an observed migration shift. Verify target identity with orthogonal controls.
Could BCL2L2 isoforms produce different bands?
Isoforms · UniProt lists isoforms 1 and 3. In isoform 3, the canonical sequence at residues 145–193 is replaced by a longer alternative sequence. The isoforms could therefore migrate differently, but the feature alone does not predict their apparent band positions. Check which isoform the antibody recognizes.
Which BCL2L2 modifications matter when interpreting bands?
PTM · The supplied modified-residue entry gives UniProt position 2 and combines N-acetylalanine, phosphoserine, and omega-N-methylarginine annotations. It does not resolve separate coordinates for the phosphorylation or methylation annotations. Keep UniProt numbering explicit when comparing antibody or paper sites; modification annotations alone do not establish a visible shift.
Does this guide establish induction of BCL2L2?
Induction · No general induction response is established by this guide. A pathway or gene-set association is not evidence of induction in a particular specimen. Verify the relevant treatment and control in a target-specific experiment.
What transfer method to use for BCL2L2 Western blot?
Transfer · BCL2L2 is a peripheral mitochondrial membrane protein with a predicted mass of 20.7 kDa. When optimizing transfer, check recovery around 21 kDa in both the gel and membrane. The supplied features do not establish a specific transfer method or settings.
How should blocking be optimized?
Blocking · Standard workflow guidance: follow the PA1426-1 datasheet where specified. Otherwise compare 5% milk or 5% BSA in TBST; for a phospho-specific assay start with BSA. Optimize background and specific signal with matched controls.
How can BCL2L2 bands be quantified across conditions?
Quantitation · Compare the same sample fraction and extraction procedure across conditions, especially when apoptosis differs, because BCL2L2 membrane association changes during apoptosis. Quantify the band assigned to the intended isoform consistently; UniProt lists isoforms 1 and 3 with different C-terminal sequences.
Why might BCL2L2 migrate near 21 kDa?
Interpretation · The canonical 193-residue sequence has a predicted mass of 20.7 kDa, close to the observed ~21 kDa band. This agreement supports the assignment but does not establish identity; confirm it with an appropriate specificity control.
BCL2L2 is loosely associated with the mitochondrial membrane in healthy cells and tightly bound during apoptosis. Keep fractionation and extraction conditions consistent across samples; a change in recovered signal may reflect altered membrane association rather than a change in total protein.
First consider antibody recognition of both listed isoforms: isoform 3 replaces canonical residues 145–193 with a longer sequence. The supplied record also lists modifications, but their presence alone cannot explain a band shift. It lists no glycosylation sites, signal peptide, or propeptide. Verify band identity before assigning a cause.
BCL2L2 is loosely associated with the mitochondrial membrane in healthy cells and tightly bound during apoptosis. Keep fractionation and extraction conditions consistent across samples; a change in recovered signal may reflect altered membrane association rather than a change in total protein.
First consider antibody recognition of both listed isoforms: isoform 3 replaces canonical residues 145–193 with a longer sequence. The supplied record also lists modifications, but their presence alone cannot explain a band shift. It lists no glycosylation sites, signal peptide, or propeptide. Verify band identity before assigning a cause.
Boster reagents
BCL2L2 Western Blot Antibodies
Catalog antibodies with Western blot application and product-specific WB images. Evaluate suitability with the reported sample, controls and experimental conditions.
Real WB data
Anti-Bcl-2-like protein 2 Bcl2L2 Antibody Picoband®
Cat # PA1426-1
The listed anti-BCL2L2 antibody, PA1426-1, is reported reactive with human, mouse, and rat. Its Western blot image shows an approximately 21 kDa band in five human cell lysates under the stated conditions; the supplied evidence does not show mouse or rat blots.
Which to pick: PA1426-1 is the only listed option and has a Western blot image using human A549, K562, U-937, HeLa, and A431 whole-cell lysates. For mouse or rat samples, catalogued reactivity is reported, but no corresponding blot is supplied.
Source: BosterBio BCL2L2 gene-info card — filtered to Western-blot-capable antibodies; each card shows that product's actual WB validation figure.
References
- UniProt Consortium. UniProt entry Q92843.
- Human Protein Atlas. BCL2L2 tissue expression.
- PMC5094470 — target-verified WB comparison
- PMC6221029 — target-verified WB comparison
- PMC6143943 — target-verified WB comparison