This website uses cookies to ensure you get the best experience on our website.
- Table of Contents
BATF3 / Basic leucine zipper transcriptional factor ATF-like 3 · Western blot design guide
Design a Western Blot for BATF3
Real validated BATF3 Western blot protocols, expected-band and isoform facts, troubleshooting for weak or shifted signal, and recommended anti-BATF3 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
BATF3 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 | ~14.5 kDa | |
| Gel | 15% (standard starting point) | |
| Negative control | Suggested KO / knockdown lysate |
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 BATF3 Western Blot Protocols
The A01957 protocol combines labelled catalog values with standard starting conditions. Published comparisons retain their own sample, reagent and detection scope.
| Sample / lysate | mouse-lung mouse-spleen lysis (catalog A01957) |
| 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 | A01957; use the WB datasheet starting dilution (standard starting point) |
| Primary incubation | Overnight at 4 °C (standard starting point) |
| Secondary antibody | 1:20000 (catalog A01957) |
| 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 BATF3 Western Blot Band Size?
BATF3 is predicted at 14.5 kDa; phosphorylation could affect migration, but no distinct band position has been demonstrated.
| Band near 14.5 kDa | consistent with the predicted size of BATF3; confirm identity with antibody controls |
| Close doublet near 14.5 kDa | could reflect phosphorylation at Ser2 or Ser31, but distinct migration is unproven |
| Weak band in cytoplasmic fraction | consistent with BATF3 localization to the nucleus |
| Stronger band in nuclear fraction | consistent with nuclear enrichment of BATF3 |
💡Expected BATF3 appearanceBATF3 has a predicted mass of 14.5 kDa, but no empirical band size is supplied; phosphorylation may affect migration, so confirm a candidate band with antibody controls.
| UniProt predicted mass | sets a 14.5 kDa reference for the full-length protein |
| 127-amino-acid sequence | underlies the predicted full-length mass; no processed size is specified |
| Ser2 phosphorylation | may alter migration, but no measurable shift is supplied |
| Ser31 phosphorylation | may alter migration, but no measurable shift is supplied |
| Situation | Likely cause | Next action |
|---|---|---|
| No band in lysate | BATF3 is nuclear and may be underrepresented in the preparation | check a nuclear extract and confirm fraction quality |
| Band higher than expected | phosphorylation at Ser2 or Ser31 may affect migration, but its effect is unmeasured | compare phosphatase-treated and untreated samples with identity controls |
| Band lower than expected | the supplied features do not establish a smaller BATF3 product | check antibody specificity and compare with full-length BATF3 |
| Multiple bands | phosphorylation may produce distinct states, but their separation is unproven | compare phosphatase-treated samples and verify band identity |
| Weak or no signal | a cytoplasmic preparation may contain little nuclear BATF3 | test a nuclear extract and verify extraction quality |
Experiment evidence
Western blot images showing BATF3 band
A01957 · BATF3
← swipe / scroll for more images →
Real WB data behind the band-size expectations in this section.
Sample controls for BATF3 Western blot
🧪For positive controls for BATF3 in Western blot, you can use no HPA-supported positive sample because none is reported.
Positive control: No high/medium HPA tissue identified
Negative control: Suggested KO / knockdown lysate
Loading controls: Run GAPDH, β-actin, and a total-protein stain alongside.
⚠️Feasibility: BATF3 is nuclear, but HPA provides no data to validate a positive or negative sample.
HPA tissue expression evidence for BATF3
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 |
|---|---|---|---|---|
| No high/medium HPA tissues identified in the supplied evidence. | ||||
Lower expression tissues · IHC evidence, not confirmed WB-negative controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| No lower-expression tissue rows available in the supplied evidence. | ||||
Section 3
Advanced BATF3 Western Blot Tips
Deeper troubleshooting and optimisation questions for BATF3, answered from its protein features.
What band size should I expect for BATF3?
Band shift · BATF3 has a predicted mass of 14.5 kDa. No observed Western blot band size is supplied, so use 14.5 kDa as a starting point rather than a confirmed apparent mass.
Could BATF3 isoforms explain multiple bands?
Isoforms · The supplied record lists one isoform and no alternative sequence. It therefore provides no isoform-based explanation for multiple bands.
Could phosphorylation shift the BATF3 band?
PTM · UniProt lists phosphoserine at residues 2 and 31. These are UniProt coordinates and may differ from antibody or paper numbering. Their presence alone does not establish a visible shift or explain a difference from 14.5 kDa.
Does this guide establish induction of BATF3?
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 BATF3 Western blot?
Transfer · BATF3 is predicted to be 14.5 kDa. Choose transfer conditions suited to retaining a small protein, and verify recovery on the membrane. The supplied features do not establish a specific transfer method.
How should blocking be optimized?
Blocking · Standard workflow guidance: follow the A01957 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 I quantify BATF3 bands?
Quantitation · Measure bands within the linear signal range and normalize against a consistent loading measure for the fraction tested. BATF3 is nuclear, so keep the sample fraction consistent across comparisons.
Which fraction should I use to detect BATF3?
Interpretation · BATF3 is annotated as nuclear. A nuclear fraction may help detect it; compare fraction loading consistently when interpreting signal.
BATF3 heterodimerizes with JUN family proteins, but that interaction alone does not identify an unexpected band. Check the band against the 14.5 kDa prediction and validate its identity; no observed band size is supplied.
BATF3 heterodimerizes with JUN family proteins, but that interaction alone does not identify an unexpected band. Check the band against the 14.5 kDa prediction and validate its identity; no observed band size is supplied.
Boster reagents
BATF3 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-BATF3/Snft Antibody
Cat # A01957
The catalog reports one anti-BATF3 antibody, A01957, with reported human, mouse, and rat reactivity. Its Western blot image shows mouse lung and spleen lysates at a 1:2000 primary antibody dilution; the supplied image does not show human or rat samples.
Which to pick: A01957 is the only listed option. Its supplied Western blot image uses mouse lung and spleen lysates, so those are the directly illustrated sample contexts. Human and rat reactivity is listed but not shown in the supplied image.
Source: BosterBio BATF3 gene-info card — filtered to Western-blot-capable antibodies; each card shows that product's actual WB validation figure.
References
- UniProt Consortium. UniProt entry Q9NR55.
- Human Protein Atlas. BATF3 tissue expression.
- PMC9004122 — target-verified WB comparison
- PMC8078141 — target-verified WB comparison
- PMC9045983 — target-verified WB comparison