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BCL11A / BCL11 transcription factor A · Western blot design guide
Design a Western Blot for BCL11A
Real validated BCL11A Western blot protocols, expected-band and isoform facts, troubleshooting for weak or shifted signal, and recommended anti-BCL11A 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
BCL11A 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 | ~91.2 kDa | |
| Observed band | ~120 kDa | |
| Gel | 8–10% (standard starting point) | |
| Positive control | Cerebellum (IHC candidate; verify WB) +4 more | |
| Negative control | Adipose tissue (IHC candidate; verify WB) |
Important caveats
Reasons your observed band may differ from the expected size.
| PTM | Phosphorylated + Methylated | |
| Caveat | Modification-state controls | |
| Gene-set association | MSigDB C7 membership | |
| Isoform | 5 isoform(s) |
Section 1
Real Curated BCL11A Western Blot Protocols
The M00741 protocol combines labelled catalog values with standard starting conditions. Published comparisons retain their own sample, reagent and detection scope.
| Sample / lysate | Jurkat cell lysate (catalog M00741) |
| Gel % | 8–10% (standard starting point) |
| Load | 20–30 µg total protein per lane; optimize for abundance (standard starting point) |
| Transfer | Standard semi-dry transfer; verify efficiency (standard starting point) |
| Membrane | 0.45 µm PVDF (standard starting point) |
| Blocking | 5% milk or 5% BSA in TBST (standard starting point) |
| Primary antibody | M00741; use the WB datasheet starting dilution (standard starting point) |
| Primary incubation | Overnight at 4 °C (standard starting point) |
| Secondary antibody | Species-matched HRP conjugate at validated dilution (standard starting point) |
| Secondary incubation | 1 h at room temperature (standard starting point) |
| Wash | 3 × 5 min in TBST (standard starting point) |
| Detection | ECL; bracket exposures to avoid saturation (standard starting point) |
Section 2
What Is the Expected BCL11A Western Blot Band Size?
BCL11A is predicted at 91.2 kDa but observed near 120 kDa; the cause of this difference is not established.
| Band near 120 kDa | Empirical BCL11A band; confirm identity with antibody and loss-of-signal controls |
| Band near 91.2 kDa | Near the predicted full-length mass; identity requires confirmation |
| Multiple discrete bands | Could reflect the five annotated isoforms; their migration differences are unestablished |
| Weak band in whole-cell lysate | Nuclear and chromosome-associated BCL11A may be underrepresented after extraction |
💡Expected BCL11A appearanceBCL11A has a predicted mass of 91.2 kDa, while an empirical blot shows a band near 120 kDa; the cause of the difference is unestablished, so confirm band identity with appropriate controls.
| Predicted full-length mass | 91.2 kDa from sequence; empirical migration is near 120 kDa |
| Splice isoform 1 | Apparent size relative to other isoforms is not supplied |
| Splice isoform 2 | May differ in size; migration relative to other isoforms is not supplied |
| Splice isoform 3 | May differ in size; migration relative to other isoforms is not supplied |
| Splice isoform 6 | May differ in size; migration relative to other isoforms is not supplied |
| Splice isoform 7 | May differ in size; migration relative to other isoforms is not supplied |
| Situation | Likely cause | Next action |
|---|---|---|
| No band in lysate | Nuclear or chromosome-associated BCL11A may be poorly extracted | Check a nuclear fraction and extraction efficiency |
| Band higher than expected | The empirical 120 kDa band exceeds the 91.2 kDa sequence prediction; the cause is unestablished | Confirm identity with an independent antibody or BCL11A depletion |
| Band lower than expected | An annotated isoform could differ in size, but isoform migration is unestablished | Check antibody epitope coverage and confirm the band by BCL11A depletion |
| Multiple bands | Five annotated isoforms could contribute, but separate bands are not demonstrated | Compare isoform expression and test which bands disappear after BCL11A depletion |
| Weak or no signal | The tested preparation may contain little extractable BCL11A | Check extraction and use a documented positive sample such as Jurkat lysate or mouse brain lysate |
Experiment evidence
Western blot images showing BCL11A band
M00741 · BCL11A
M00741 · BCL11A
← swipe / scroll for more images →
Real WB data behind the band-size expectations in this section.
Sample controls for BCL11A Western blot
🧪HPA-IHC candidate guidance (verify in WB): For positive controls for BCL11A in Western blot, you can use cerebellum lysate, which has high HPA expression.
Positive control: Cerebellum (IHC candidate; verify WB)
Negative control: Adipose tissue (IHC candidate; verify WB)
Loading controls: Run GAPDH, β-actin, and a total-protein stain alongside the samples.
⚠️Feasibility: Adipose tissue is HPA not detected, but its lack of Western-blot signal needs validation.
HPA tissue expression evidence for BCL11A
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 |
|---|---|---|---|---|
| Cerebellum | Purkinje cells | High | Protein (IHC) | HPA → |
| Cerebral cortex | neuronal cells | High | Protein (IHC) | HPA → |
| Hippocampus | neuronal cells | High | Protein (IHC) | HPA → |
| Appendix | lymphoid tissue | Medium | Protein (IHC) | HPA → |
| Cervix | squamous epithelial cells | Medium | Protein (IHC) | HPA → |
Lower expression tissues · IHC evidence, not confirmed WB-negative controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| Adipose tissue | adipocytes | Not detected | Protein (IHC) | HPA → |
| Adrenal gland | glandular cells | Not detected | Protein (IHC) | HPA → |
| Colon | endothelial cells | Not detected | Protein (IHC) | HPA → |
| Duodenum | glandular cells | Not detected | Protein (IHC) | HPA → |
| Endometrium | cells in endometrial stroma | Not detected | Protein (IHC) | HPA → |
Section 3
Advanced BCL11A Western Blot Tips
Deeper troubleshooting and optimisation questions for BCL11A, answered from its protein features.
How should BCL11A 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.
Which BCL11A isoforms could produce different bands?
Isoforms · Five isoforms are listed. Relative to the 835-residue canonical sequence, isoform 6 changes residues 129–163, isoform 7 lacks 143–835, isoform 3 lacks 244–835, and isoform 2 lacks 774–835. Their sequence differences could affect band size and antibody recognition; a second band is not proof of an isoform.
Choose an epitope using the sequence differences relevant to your question. A region within canonical residues 774–835 is absent from isoforms 2, 3, and 7; a region within 244–744 is absent from isoforms 3 and 7. Check the exact epitope against each isoform sequence before interpreting a missing signal.
Choose an epitope using the sequence differences relevant to your question. A region within canonical residues 774–835 is absent from isoforms 2, 3, and 7; a region within 244–744 is absent from isoforms 3 and 7. Check the exact epitope against each isoform sequence before interpreting a missing signal.
Which BCL11A modifications should inform band interpretation?
PTM · UniProt lists phosphoserine at 86, 205, 332, 337, 446, 447, 608, 625, and 630; phosphothreonine at 701; and asymmetric dimethylarginine at 271. These are canonical UniProt coordinates, which may differ from paper or antibody numbering. Their presence does not establish a visible shift.
Does this guide establish induction of BCL11A?
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 BCL11A Western blot?
Transfer · With an observed band near 120 kDa, check that your transfer conditions retain and move proteins in that size range. Inspect the membrane and post-transfer gel around 120 kDa before interpreting a weak signal; the supplied features do not specify a single transfer method.
How should blocking be optimized?
Blocking · Standard workflow guidance: follow the M00741 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 BCL11A bands be quantified across samples?
Quantitation · BCL11A is reported in the cytoplasm, nucleus, and on chromosomes. Compare matched sample fractions and quantify the same defined band or bands across samples. If multiple bands appear, report their intensities separately unless their identities have been established.
Why might BCL11A appear near 120 kDa instead of 91.2 kDa?
Interpretation · The supplied observed band is about 120 kDa, while the predicted mass is 91.2 kDa. BCL11A has modified residues and alternative isoforms, but those features alone do not explain the difference or prove a band shift. Confirm the band with an antibody recognizing a known BCL11A region.
Check whether the antibody epitope is retained in the candidate isoform. Isoforms 3 and 7 lack long C-terminal regions, while isoform 2 lacks residues 774–835. BCL11A also self-associates, but that feature alone does not identify an unexpected band or establish its apparent mass.
Check whether the antibody epitope is retained in the candidate isoform. Isoforms 3 and 7 lack long C-terminal regions, while isoform 2 lacks residues 774–835. BCL11A also self-associates, but that feature alone does not identify an unexpected band or establish its apparent mass.
Boster reagents
BCL11A 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-Ctip1 Monoclonal Antibody
Cat # M00741
The catalog reports M00741, an anti-Ctip1 monoclonal antibody for BCL11A with reported human, mouse, and rat reactivity. Its WB image shows Ctip1 expression in Jurkat cell lysate; no other tested samples are supplied.
Which to pick: M00741 is the only listed option. Its WB image uses Jurkat cell lysate; check whether that sample context and the reported reactivity fit your experiment.
Source: BosterBio BCL11A gene-info card — filtered to Western-blot-capable antibodies; each card shows that product's actual WB validation figure.
References
- UniProt Consortium. UniProt entry Q9H165.
- Human Protein Atlas. BCL11A tissue expression.
- PMC9391307 — target-verified WB comparison
- PMC7098986 — target-verified WB comparison
- PMC8530301 — target-verified WB comparison