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BPTF / Nucleosome-remodeling factor subunit BPTF · Western blot design guide
Design a Western Blot for BPTF
Source-linked BPTF Western blot protocol options, expected-band and isoform facts, troubleshooting for weak or shifted signal, and recommended anti-BPTF 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
BPTF Western Blot Experimental Design Guide
Expected bands, source-linked protocol options, controls and antibodies — the at-a-glance facts below, then the full design guide.
Must know before running
| Expected band | ~338.3 kDa | |
| Gel | 4–12% gradient (standard starting point) | |
| Positive control | Testis (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 + Acetylated | |
| Caveat | Phosphorylation-state controls | |
| Gene-set association | MSigDB C7 membership | |
| Isoform | 3 isoform(s) |
Section 1
Source-Linked BPTF Western Blot Protocol Options
The A03754 protocol combines labelled catalog values with standard starting conditions. Published comparisons retain their own sample, reagent and detection scope.
| Sample / lysate | Target-positive lysate and matched negative control (standard starting point) |
| Gel % | 4–12% gradient (standard starting point) |
| Load | 20–30 µg total protein per lane; optimize for abundance (standard starting point) |
| Transfer | Wet/tank transfer; optimize duration (standard starting point) |
| Membrane | 0.45 µm PVDF (standard starting point) |
| Blocking | 5% milk or 5% BSA in TBST (standard starting point) |
| Primary antibody | A03754; 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 BPTF Western Blot Band Size?
Full-length BPTF is predicted at 338.3 kDa; isoforms and modifications are documented, but their effects on observed migration are not demonstrated.
| Band near 338.3 kDa | Consistent with the predicted size of full-length BPTF; identity requires confirmation. |
| Multiple bands at different sizes | Isoforms 1, 2, and 4 could contribute, but their migration is not established. |
| Band with slightly altered mobility | Reported phosphorylation or acetylation could contribute; a visible shift is not established. |
| Weak band in a cytoplasmic fraction | BPTF is also found in the nucleus. |
💡Expected BPTF appearanceFull-length BPTF has a predicted mass of 338.3 kDa, but no empirical band size is supplied; confirm any candidate band with a positive control and an independent antibody or BPTF depletion.
| UniProt predicted mass | Places full-length BPTF at approximately 338.3 kDa before considering migration differences. |
| Isoforms 1, 2, and 4 | May differ in size, but their relative masses and separation on a blot are not supplied. |
| Ser216 phosphorylation | Could affect mobility; no visible size shift is established. |
| Thr1064 phosphorylation | Could affect mobility; no visible size shift is established. |
| Lys880 acetylation | Could affect mobility; no visible size shift is established. |
| Situation | Likely cause | Next action |
|---|---|---|
| No band in lysate | Nuclear BPTF may be poorly recovered in the preparation. | Check a nuclear extract and a known positive lysate. |
| Band higher than expected | Its identity or a modification-related mobility change is uncertain. | Check a high-mass marker and confirm with an independent antibody or BPTF depletion. |
| Band lower than expected | An isoform or a BPTF fragment is possible; neither has a supplied band size. | Compare antibodies recognizing different regions and use BPTF depletion. |
| Multiple bands | Isoforms or reported modifications could contribute, but distinct bands are not established. | Use BPTF depletion and compare phosphatase-treated and untreated samples. |
| Weak or no signal | The large BPTF protein may transfer inefficiently. | Check transfer of high-mass markers and test a positive lysate. |
| Fragments below expected size | The long BPTF protein may undergo degradation during sample preparation. | Prepare fresh lysate with protease inhibitors and compare antibodies recognizing different regions. |
Experiment evidence
Western blot images showing BPTF band
A03754 · BPTF
← swipe / scroll for more images →
Real WB data behind the band-size expectations in this section.
Sample controls for BPTF Western blot
🧪HPA-IHC candidate guidance (verify in WB): For positive controls for BPTF in Western blot, you can use testis lysate, which has 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 such as Ponceau alongside the samples.
⚠️Feasibility: BPTF is intracellular, and HPA reports adipose tissue as not detected, making tissue lysates feasible controls.
HPA tissue expression evidence for BPTF
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 | spermatogonia cells | High | Protein (IHC) | HPA → |
| Adrenal gland | glandular cells | Medium | Protein (IHC) | HPA → |
| Caudate | neuronal cells | Medium | Protein (IHC) | HPA → |
| Cerebellum | Purkinje cells | Medium | Protein (IHC) | HPA → |
| Colon | glandular 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 → |
| Bone marrow | hematopoietic cells | Not detected | Protein (IHC) | HPA → |
| Cervix | glandular cells | Not detected | Protein (IHC) | HPA → |
| Epididymis | glandular cells | Not detected | Protein (IHC) | HPA → |
| Esophagus | squamous epithelial cells | Not detected | Protein (IHC) | HPA → |
Section 3
Advanced BPTF Western Blot Tips
Deeper troubleshooting and optimisation questions for BPTF, answered from its protein features.
How should BPTF 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.
How could BPTF isoforms affect band interpretation?
Isoforms · Isoforms 1, 2 and 4 are listed. Relative to the canonical UniProt sequence, isoform 2 lacks residues 622–747 and isoform 4 lacks residues 2522–2664. Check whether the antibody epitope falls within either missing segment before assigning a band to an isoform. These sequence differences do not establish where bands will migrate.
Which BPTF phosphorylation sites matter when interpreting bands?
PTM · UniProt lists phosphoserine at 216, 572, 763, 817, 1231, 1300, 1310, 2098 and 2465, and phosphothreonine at 1064 and 1303. These are UniProt canonical-sequence coordinates; antibody or paper numbering may differ. Their presence does not demonstrate a visible band shift.
UniProt lists N6-acetyllysine at 880, omega-N-methylarginine at 2155, and asymmetric dimethylarginine at 2162, 2184 and 2191. Check whether the antibody recognizes an epitope containing a listed site, especially when comparing modified and total BPTF signals. Coordinates refer to the UniProt canonical sequence.
UniProt lists N6-acetyllysine at 880, omega-N-methylarginine at 2155, and asymmetric dimethylarginine at 2162, 2184 and 2191. Check whether the antibody recognizes an epitope containing a listed site, especially when comparing modified and total BPTF signals. Coordinates refer to the UniProt canonical sequence.
Does this guide establish induction of BPTF?
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 BPTF Western blot?
Transfer · BPTF has a predicted mass of 338.3 kDa, so assess transfer of high-mass proteins and check signal remaining in the gel. The supplied UniProt features do not identify a particular membrane, transfer mode or setting.
How should blocking be optimized?
Blocking · Standard workflow guidance: follow the A03754 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 BPTF bands be quantified across samples?
Quantitation · Use the same antibody and quantify a consistently defined band or set of bands. Record whether samples are whole-cell, nuclear or cytoplasmic: UniProt lists BPTF in both nucleus and cytoplasm and notes localization to DNA-damage sites. Its three isoforms may also complicate comparisons if the antibody recognizes them differently.
Why might BPTF migrate differently from its predicted mass?
Interpretation · The predicted mass is 338.3 kDa. BPTF has three listed isoforms and multiple modified residues, but these features alone do not establish an apparent mass or explain any observed difference. Compare the band with a high-mass marker and check which isoform the antibody recognizes.
First check antibody epitope coverage against the isoform 2 deletion at UniProt residues 622–747 and the isoform 4 deletion at 2522–2664. Then assess whether the band is consistent across samples and whether high-mass transfer is adequate. The listed modifications and isoforms are possibilities to examine, not proof of a particular band identity.
First check antibody epitope coverage against the isoform 2 deletion at UniProt residues 622–747 and the isoform 4 deletion at 2522–2664. Then assess whether the band is consistent across samples and whether high-mass transfer is adequate. The listed modifications and isoforms are possibilities to examine, not proof of a particular band identity.
Boster reagents
BPTF 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-BPTF/Falz Monoclonal Antibody
Cat # A03754
The catalog reports one anti-BPTF monoclonal antibody, A03754, with stated human reactivity. Its Western blot image uses HEK293 lysate and BPTF (AA 503–670)-hIgGFc transfected HEK293 lysate. The supplied evidence is limited to those reported samples.
Which to pick: A03754 is the only listed antibody and has a Western blot image. Use its reported HEK293 comparison as the starting reference; performance in other samples or conditions is not shown.
Source: BosterBio BPTF gene-info card — filtered to Western-blot-capable antibodies; each card shows that product's actual WB validation figure.
References
- UniProt Consortium. UniProt entry Q12830.
- Human Protein Atlas. BPTF tissue expression.
- PMC10176855 — target-verified WB comparison
- PMC4741809 — target-verified WB comparison
- PMC5093041 — target-verified WB comparison