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- Table of Contents
BATF / Basic leucine zipper transcriptional factor ATF-like · Western blot design guide
Design a Western Blot for BATF
Real validated BATF Western blot protocols, expected-band and isoform facts, troubleshooting for weak or shifted signal, and recommended anti-BATF 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
BATF 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 | ~14.1 kDa | |
| Gel | 15% (standard starting point) | |
| Positive control | Testis (IHC candidate; verify WB) | |
| Negative control | Adipose tissue (IHC candidate; verify WB) |
Important caveats
Reasons your observed band may differ from the expected size.
| PTM | Phosphorylated | |
| Caveat | Phosphorylation-state controls | |
| Gene-set association | MSigDB Hallmark membership | |
| Isoform | 1 isoform(s) |
Section 1
Real Curated BATF Western Blot Protocols
The A04089 protocol combines labelled catalog values with standard starting conditions. Published comparisons retain their own sample, reagent and detection scope.
| Sample / lysate | rat spleen tissue lysate (catalog A04089) |
| Gel % | 15% (standard starting point) |
| Load | 20–30 µg total protein per lane; optimize for abundance (standard starting point) |
| Transfer | Short semi-dry transfer; verify retention (standard starting point) |
| Membrane | 0.2 µm PVDF (standard starting point) |
| Blocking | 5% milk or 5% BSA in TBST (standard starting point) |
| Primary antibody | A04089 · 1 μg/mL (catalog A04089) |
| 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 BATF Western Blot Band Size?
BATF is predicted at 14.1 kDa; phosphorylation and localization may affect interpretation, but altered migration has not been demonstrated.
| Band near 14.1 kDa | Consistent with the predicted BATF monomer; confirm identity with antibody controls |
| Slightly shifted band near 14.1 kDa | Phosphorylation at Ser43 or Thr48 is possible, but a mobility shift is not established |
| Band in nuclear and cytoplasmic fractions | Consistent with BATF localization in both compartments |
| Stronger nuclear band after T-cell activation | Consistent with increased nuclear translocation |
💡Expected BATF appearanceBATF has a predicted mass of 14.1 kDa, but no empirical band size is supplied; confirm any band near that size with antibody and sample controls.
| Predicted BATF mass | Places the monomer near 14.1 kDa |
| Ser43 phosphorylation | Could affect mobility; no shift is demonstrated |
| Thr48 phosphorylation | Could affect mobility; no shift is demonstrated |
| BATF–JUNB heterodimerization | Does not establish a higher band under denaturing conditions |
| Situation | Likely cause | Next action |
|---|---|---|
| No band in lysate | Nuclear BATF may be poorly recovered | Check nuclear extraction and a positive sample |
| Band higher than expected | Identity and any phosphorylation-related shift are unconfirmed | Check antibody specificity and compare phosphatase-treated material |
| Band lower than expected | Band identity is uncertain; no cleavage feature is listed | Check antibody specificity and sample integrity |
| Multiple bands | No distinct isoforms or migration pattern are established | Use antibody specificity controls and compare phosphatase-treated material |
| Weak or no signal | BATF distribution can change after T-cell activation | Compare nuclear and cytoplasmic fractions with fraction controls |
Experiment evidence
Western blot images showing BATF band
A04089 · BATF
← swipe / scroll for more images →
Real WB data behind the band-size expectations in this section.
Sample controls for BATF Western blot
🧪HPA-IHC candidate guidance (verify in WB): For positive controls for BATF in Western blot, you can use testis lysate, based on its high HPA expression.
Positive control: Testis (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: An HPA not-detected tissue provides a negative control, but BATF localization can shift toward the nucleus after T-cell activation.
HPA tissue expression evidence for BATF
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 |
|---|---|---|---|---|
| Testis | sertoli cells | High | 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 → |
| Bone marrow | hematopoietic cells | Not detected | Protein (IHC) | HPA → |
| Breast | adipocytes | Not detected | Protein (IHC) | HPA → |
| Bronchus | respiratory epithelial cells | Not detected | Protein (IHC) | HPA → |
Section 3
Advanced BATF Western Blot Tips
Deeper troubleshooting and optimisation questions for BATF, answered from its protein features.
How should BATF 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 BATF isoforms account for multiple bands?
Isoforms · The supplied record lists one isoform and no alternative sequence. It provides no annotated isoform explanation for multiple bands.
Which BATF phosphorylation sites matter when interpreting bands?
PTM · UniProt lists phosphoserine at position 43 and phosphothreonine at position 48, using UniProt sequence numbering. Their presence alone does not establish a visible band shift or explain a mass difference.
Does this guide establish induction of BATF?
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.
How should transfer be checked for BATF?
Transfer · Standard workflow guidance: verify transfer efficiency for the intended target size before interpreting a weak signal. Use total-protein assessment and optimize transfer for the membrane, gel and apparatus; the labelled catalog values take precedence.
How should blocking be optimized?
Blocking · Standard workflow guidance: follow the A04089 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 BATF be quantified across cell fractions?
Quantitation · Because BATF occurs in both nucleus and cytoplasm and shifts toward the nucleus after T-cell activation, quantify comparable fractions under matched conditions. A change in nuclear signal alone does not establish a change in total BATF abundance.
Should BATF appear exactly at its predicted 14.1 kDa?
Interpretation · 14.1 kDa is the calculated mass of the 125-residue BATF sequence. No observed band position is supplied, so an apparent mass or shift cannot be predicted from these features.
BATF is listed in the nucleus and cytoplasm, with increased nuclear translocation after T-cell activation. Compare like fractions when assessing activation; this feature does not establish increased total BATF expression.
BATF mainly heterodimerizes with JUNB, and the complex interacts with IRF4 and IRF8. Those interactions do not establish that a higher band contains the complex. The supplied features do not identify an observed band.
BATF is listed in the nucleus and cytoplasm, with increased nuclear translocation after T-cell activation. Compare like fractions when assessing activation; this feature does not establish increased total BATF expression.
BATF mainly heterodimerizes with JUNB, and the complex interacts with IRF4 and IRF8. Those interactions do not establish that a higher band contains the complex. The supplied features do not identify an observed band.
Boster reagents
BATF 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-BATF Antibody
Cat # A04089
The catalog reports one anti-BATF antibody, A04089, with reported human, mouse, and rat reactivity. Its WB image shows rat spleen tissue lysate tested at 1 μg/mL; the supplied evidence does not show WB results for human or mouse samples.
Which to pick: A04089 is the only listed option. Its documented WB example uses rat spleen lysate at 1 μg/mL; confirm performance in your own sample type and conditions.
Source: BosterBio BATF gene-info card — filtered to Western-blot-capable antibodies; each card shows that product's actual WB validation figure.
References
- UniProt Consortium. UniProt entry Q16520.
- Human Protein Atlas. BATF tissue expression.
- PMC12515186 — target-verified WB comparison
- PMC11165778 — target-verified WB comparison
- PMC12593631 — target-verified WB comparison