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- Table of Contents
BCLAF1 / Bcl-2-associated transcription factor 1 · Western blot design guide
Design a Western Blot for BCLAF1
Real validated BCLAF1 Western blot protocols, expected-band and isoform facts, troubleshooting for weak or shifted signal, and recommended anti-BCLAF1 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
BCLAF1 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 | ~106.1 kDa | |
| Observed band | ≈150 kDa | |
| Gel | 8–10% (standard starting point) | |
| Positive control | Adrenal gland (IHC candidate; verify WB) +4 more | |
| Negative control | Suggested KO / knockdown lysate |
Important caveats
Reasons your observed band may differ from the expected size.
| PTM | Phosphorylated + Acetylated | |
| Caveat | Band identity controls | |
| Gene-set association | MSigDB C7 membership | |
| Isoform | 4 isoform(s) |
Section 1
Real Curated BCLAF1 Western Blot Protocols
The A04100-3 protocol combines labelled catalog values with standard starting conditions. Published comparisons retain their own sample, reagent and detection scope.
| Sample / lysate | Hela (30ug), Rabbit control IgG instead of anti-BCLAF1 antibody in Hela, anti-BCLAF1 antibody (2μg) + Hela (500μg) (catalog A04100-3) |
| Gel % | 8–10% (standard starting point) |
| Load | 20–30 µg total protein per lane; optimize for abundance (standard starting point) |
| Transfer | a membrane (catalog A04100-3) |
| Membrane | 0.45 µm PVDF (standard starting point) |
| Blocking | 5% milk or 5% BSA in TBST (standard starting point) |
| Primary antibody | A04100-3 · 0.5 μg/mL (catalog A04100-3) |
| Primary incubation | Overnight at 4 °C (standard starting point) |
| Secondary antibody | goat anti-rabbit IgG-HRP (catalog A04100-3) |
| Secondary incubation | 1 h at room temperature (standard starting point) |
| Wash | 3 × 5 min in TBST (standard starting point) |
| Detection | ECL (catalog A04100-3) |
Section 2
What Is the Expected BCLAF1 Western Blot Band Size?
BCLAF1 is predicted at 106.1 kDa and observed near 150 kDa in antibody QC blots; the cause of the difference is not established.
| Band near 150 kDa | Empirical BCLAF1 band in whole-cell lysate; confirm identity with controls |
| Band near 106 kDa | Near the predicted sequence mass; identity requires confirmation |
| Several bands at different positions | Could include BCLAF1 isoforms 1, 2, 3, and 4; their migration is unreported |
| Weak band in a cytoplasmic fraction | BCLAF1 is also found in the nucleus |
💡Expected BCLAF1 appearanceBCLAF1 has a predicted mass of 106.1 kDa, while antibody QC shows a band at approximately 150 kDa in whole-cell lysate; confirm band identity with ordinary specificity controls.
| Predicted sequence mass | 106.1 kDa predicted; the empirical whole-cell band is approximately 150 kDa, with the difference unexplained |
| Splice isoforms 1 and 2 | their relative apparent sizes are not supplied |
| Splice isoform 3 | its apparent size relative to other isoforms is not supplied |
| Splice isoform 4 | its apparent size relative to other isoforms is not supplied |
| Situation | Likely cause | Next action |
|---|---|---|
| No band in lysate | BCLAF1 may be poorly recovered from a preparation despite its nuclear and cytoplasmic localization | Check extraction and loading, then examine a nuclear fraction |
| Band higher than expected | The documented approximately 150 kDa band exceeds the 106.1 kDa prediction; its cause is unestablished | Compare with the documented band and confirm identity using BCLAF1 depletion or an independent antibody |
| Band lower than expected | A BCLAF1 isoform is possible, but isoform band positions are unreported | Check antibody epitope coverage and confirm the band with BCLAF1 depletion |
| Multiple bands | Four BCLAF1 isoforms are listed, but distinct bands have not been established | Compare bands after BCLAF1 depletion and with an independent antibody |
| Weak or no signal | Nuclear BCLAF1 may be underrepresented in the tested fraction | Check loading and extraction, and compare with a nuclear fraction |
Experiment evidence
Western blot images showing BCLAF1 band
A04100-3 · BCLAF1
A04100-3 · BCLAF1
A04100-1 · BCLAF1
← swipe / scroll for more images →
Real WB data behind the band-size expectations in this section.
Sample controls for BCLAF1 Western blot
🧪HPA-IHC candidate guidance (verify in WB): For positive controls for BCLAF1 in Western blot, you can use adrenal gland lysate.
Positive control: Adrenal gland (IHC candidate; verify WB)
Negative control: Suggested KO / knockdown lysate
Loading controls: Run GAPDH, β-actin, and a total-protein stain alongside the samples.
⚠️Feasibility: No clean negative tissue is identified in the supplied HPA data; use siRNA knockdown or a KO line.
HPA tissue expression evidence for BCLAF1
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 |
|---|---|---|---|---|
| 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 → |
Lower expression tissues · IHC evidence, not confirmed WB-negative controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| Adipose tissue | adipocytes | Medium | Protein (IHC) | HPA → |
| Soft tissue | fibroblasts | Medium | Protein (IHC) | HPA → |
| Adrenal gland | glandular cells | High | Protein (IHC) | HPA → |
| Appendix | glandular cells | High | Protein (IHC) | HPA → |
| Bone marrow | hematopoietic cells | High | Protein (IHC) | HPA → |
Section 3
Advanced BCLAF1 Western Blot Tips
Deeper troubleshooting and optimisation questions for BCLAF1, answered from its protein features.
What should I check if extra BCLAF1 bands appear?
Band shift · Check antibody epitope coverage against the listed isoform deletions, especially residues 339–511 in isoform 4 and 800–848 in isoform 3. Compare the extra bands with a BCLAF1-specific control. UniProt lists alternative splicing and many modifications, but their presence alone cannot identify an unexpected band.
Could BCLAF1 isoforms produce different bands?
Isoforms · Yes. UniProt lists four isoforms. Relative to isoform 1, isoforms 2 and 3 lack residues 35–36, isoform 3 also lacks 800–848, and isoform 4 lacks 339–511. Check whether the antibody epitope falls in a deleted region; the listed differences do not establish which isoform produced a band.
Which BCLAF1 modifications matter when interpreting bands?
PTM · UniProt annotates 53 modified residues, including phosphorylation, acetylation, citrullination at residue 803, and an annotation at residue 809 reading ‘Omega-N-methylarginine Phosphoserine.’ These are UniProt coordinates; antibody or paper numbering may differ. Modification annotations alone do not establish a visible shift.
BCLAF1 has numerous annotated phosphosites, including serines 102 and 104 and threonine 661 in UniProt numbering. For a site-specific antibody, compare blocking conditions using samples with different phosphorylation states and assess background alongside the target signal. Match the antibody’s stated site to UniProt numbering before interpreting the result.
BCLAF1 has numerous annotated phosphosites, including serines 102 and 104 and threonine 661 in UniProt numbering. For a site-specific antibody, compare blocking conditions using samples with different phosphorylation states and assess background alongside the target signal. Match the antibody’s stated site to UniProt numbering before interpreting the result.
Does this guide establish induction of BCLAF1?
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 BCLAF1 Western blot?
Transfer · Plan transfer assessment around the approximately 150 kDa observed band as well as BCLAF1’s 106.1 kDa predicted mass. Check whether protein near 150 kDa transfers efficiently using a total-protein stain or molecular-weight marker, and adjust transfer conditions if it does not. The supplied UniProt features do not specify a transfer method.
How should blocking be optimized?
Blocking · Standard workflow guidance: follow the A04100-3 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 should BCLAF1 bands be quantified?
Quantitation · Measure the same validated band across samples and normalize to total protein in the relevant fraction. UniProt places BCLAF1 in the cytoplasm and nucleus, including nuclear speckles and nucleoplasm, so keep fractionation consistent. If comparing isoforms, first verify that the antibody detects the isoforms being quantified.
Does the 150 kDa band match BCLAF1’s predicted mass?
Interpretation · UniProt predicts 106.1 kDa, while the supplied Western blot observation is approximately 150 kDa. BCLAF1 has many annotated modifications and four isoforms, but those features alone do not explain the difference. Confirm the band with a BCLAF1-specific control before assigning it.
Boster reagents
BCLAF1 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-BCLAF1 Antibody Picoband®
Cat # A04100-3
Real WB data
Anti-BCLAF1 Antibody
Cat # A04100-1
The catalog reports two anti-BCLAF1 antibodies with reported human, mouse, and rat reactivity. A04100-1 has a WB example in human brain lysate. A04100-3 has HeLa lysate and immunoprecipitation lanes; its reported band is approximately 150 kDa versus an expected 106 kDa.
Which to pick: Choose A04100-1 for a direct WB example in human brain lysate (1 μg/mL). A04100-3 shows HeLa whole-cell lysate and immunoprecipitation lanes (0.5 μg/mL detection); assess its approximately 150 kDa band against the expected 106 kDa size.
Source: BosterBio BCLAF1 gene-info card — filtered to Western-blot-capable antibodies; each card shows that product's actual WB validation figure.
References
- UniProt Consortium. UniProt entry Q9NYF8.
- Human Protein Atlas. BCLAF1 tissue expression.
- PMC10700218 — target-verified WB comparison
- PMC13292053 — target-verified WB comparison
- PMC10858942 — target-verified WB comparison