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- Table of Contents
FUBP1 / Far upstream element-binding protein 1 · Western blot design guide
Design a Western Blot for FUBP1
Source-linked FUBP1 Western blot protocol options, expected-band and isoform facts, troubleshooting for weak or shifted signal, and recommended anti-FUBP1 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
FUBP1 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 | ~67.6 kDa | |
| Observed band | ~74 kDa | |
| Gel | 10% (catalog A03126-2) | |
| Positive control | Adipose tissue (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 | Phosphorylation-state controls | |
| Gene-set association | MSigDB C7 membership | |
| Isoform | 2 isoform(s) |
Section 1
Source-Linked FUBP1 Western Blot Protocol Options
The A03126-2 protocol combines labelled catalog values with standard starting conditions. Published comparisons retain their own sample, reagent and detection scope.
| Sample / lysate | human Jurkat, human Hela, human A431, human K562, rat brain, rat PC-12, mouse brain, mouse Neuro-2a (catalog A03126-2) |
| Gel % | 10% (catalog A03126-2) |
| Load | 30 ug; reducing conditions (catalog A03126-2) |
| Transfer | a nitrocellulose membrane at 150 mA for 50-90 minutes (catalog A03126-2) |
| Membrane | nitrocellulose membrane (catalog A03126-2) |
| Blocking | 5% non-fat milk/TBS for 1.5 hour at RT (catalog A03126-2) |
| Primary antibody | A03126-2 · 0.5 μg/mL (catalog A03126-2) |
| Primary incubation | overnight at 4°C (catalog A03126-2) |
| Secondary antibody | goat anti-rabbit IgG-HRP, 1:5000 (catalog A03126-2) |
| Secondary incubation | 1.5 hour at RT (catalog A03126-2) |
| Wash | TBS-0.1%Tween 3 times with 5 minutes each (catalog A03126-2) |
| Detection | ECL (catalog A03126-2) |
Section 2
What Is the Expected FUBP1 Western Blot Band Size?
FUBP1 is predicted at 67.6 kDa and observed near 74 kDa; the cause of the difference is not established.
| Band near 74 kDa | empirical FUBP1 band in reducing whole-cell lysates; confirm identity with controls |
| Band near 67.6 kDa | near the sequence-predicted mass; identity requires confirmation |
| Several bands at different positions | isoforms 1 and 2 are reported, but distinct migration is unestablished |
| Weak band in a cytoplasmic fraction | FUBP1 is annotated as nuclear |
💡Expected FUBP1 appearanceFUBP1 has a predicted mass of 67.6 kDa and an empirical band near 74 kDa; the reason for the difference is unestablished, so confirm band identity with ordinary controls.
| Predicted molecular mass | 67.6 kDa from the UniProt sequence; apparent migration can differ |
| Alternative splicing | may change protein size; distinct band positions are unestablished |
| Isoform 1 | reported isoform with no supplied individual mass or migration |
| Isoform 2 | reported isoform with no supplied individual mass or migration |
| Situation | Likely cause | Next action |
|---|---|---|
| No band in lysate | nuclear FUBP1 may be poorly recovered | check nuclear extraction and include a positive-control lysate |
| Band higher than expected | the observed 74 kDa band exceeds the 67.6 kDa prediction for an unestablished reason | compare with a positive control and confirm identity by FUBP1 depletion |
| Band lower than expected | identity is uncertain because isoform masses and migration are unsupplied | confirm identity by FUBP1 depletion and compare with a positive control |
| Multiple bands | isoforms 1 and 2 are reported, but separate bands are unverified | test band identity by FUBP1 depletion |
| Weak or no signal | low recovery of nuclear FUBP1 is possible | check nuclear extraction and compare with a positive-control lysate |
Experiment evidence
Western blot images showing FUBP1 band
A03126-2 · FUBP1
M03126 · FUBP1
← swipe / scroll for more images →
Real WB data behind the band-size expectations in this section.
Sample controls for FUBP1 Western blot
🧪HPA-IHC candidate guidance (verify in WB): For positive controls for FUBP1 in Western blot, you can use adipose tissue lysate.
Positive control: Adipose tissue (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 not-detected tissue is listed, so use siRNA knockdown or a KO line as the negative control.
HPA tissue expression evidence for FUBP1
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 |
|---|---|---|---|---|
| Adipose tissue | adipocytes | High | 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 → |
| Breast | glandular cells | High | Protein (IHC) | HPA → |
Lower expression tissues · IHC evidence, not confirmed WB-negative controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| Adipose tissue | adipocytes | High | 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 → |
| Breast | glandular cells | High | Protein (IHC) | HPA → |
Section 3
Advanced FUBP1 Western Blot Tips
Deeper troubleshooting and optimisation questions for FUBP1, answered from its protein features.
How should FUBP1 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 FUBP1 isoforms produce different bands?
Isoforms · UniProt lists isoforms 1 and 2. Relative to the canonical sequence, isoform 2 lacks residue 97 and replaces GQ at positions 643–644 with CRFDPASIELAL. Antibody epitope coverage may therefore matter, but these sequence changes do not establish where either isoform migrates.
Which FUBP1 phosphorylation sites matter when assessing band changes?
PTM · The supplied UniProt features list phosphoserine at 52, 55, 140 and 630, and phosphothreonine at 153 and 432. These are UniProt canonical-sequence coordinates; paper or antibody numbering may differ. A phosphorylation-dependent signal is possible, but the sites alone do not demonstrate a visible shift.
UniProt lists N-acetylalanine at position 2 and omega-N-methylarginine at 321, 359, 361 and 363. These positions use UniProt canonical-sequence numbering. The listed modifications can guide site-specific antibody interpretation, but their presence does not identify an unexpected band or account for the apparent mass.
FUBP1 has documented phosphoserine and phosphothreonine sites, so assess background and target signal when selecting a blocker for a site-specific antibody. The supplied features do not establish which blocker works best; follow the antibody's instructions and compare conditions empirically.
UniProt lists N-acetylalanine at position 2 and omega-N-methylarginine at 321, 359, 361 and 363. These positions use UniProt canonical-sequence numbering. The listed modifications can guide site-specific antibody interpretation, but their presence does not identify an unexpected band or account for the apparent mass.
FUBP1 has documented phosphoserine and phosphothreonine sites, so assess background and target signal when selecting a blocker for a site-specific antibody. The supplied features do not establish which blocker works best; follow the antibody's instructions and compare conditions empirically.
Does this guide establish induction of FUBP1?
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 FUBP1 Western blot?
Transfer · Use the approximately 74 kDa observed band to check transfer performance: inspect the membrane and post-transfer gel, then adjust conditions if target-sized protein remains in the gel. The supplied UniProt features do not specify a validated transfer method for FUBP1.
How should blocking be optimized?
Blocking · Standard workflow guidance: follow the A03126-2 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 can I quantify FUBP1 bands consistently?
Quantitation · FUBP1 is listed as nuclear and has two isoforms. Use a consistent sample fraction and antibody across comparisons, and define which band or bands are included before quantifying. If measuring a modified site, report that signal separately from total FUBP1.
Why does FUBP1 appear near 74 kDa instead of 67.6 kDa?
Interpretation · 67.6 kDa is the predicted mass; approximately 74 kDa is the reported apparent band. FUBP1 has documented modifications and two isoforms, but those features alone do not establish the cause of this difference. Use the observed position as a reference and assess band identity independently.
Check whether the antibody recognizes sequence retained in both isoforms, particularly around canonical residue 97 and positions 643–644. Compare bands across consistent nuclear samples. UniProt documents isoforms and modifications, but does not establish that any particular extra band is an isoform, modified FUBP1 or a binding partner.
Check whether the antibody recognizes sequence retained in both isoforms, particularly around canonical residue 97 and positions 643–644. Compare bands across consistent nuclear samples. UniProt documents isoforms and modifications, but does not establish that any particular extra band is an isoform, modified FUBP1 or a binding partner.
Boster reagents
FUBP1 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-FUBP1 Antibody Picoband®
Cat # A03126-2
Real WB data
Anti-FUBP1 Rabbit Monoclonal Antibody
Cat # M03126
Both listed anti-FUBP1 antibodies have Western blot images using human, mouse, and rat samples. Each reports a band near 74 kDa, above the stated expected size of 67–68 kDa. The supplied evidence does not establish band identity independently.
Which to pick: Both A03126-2 and M03126 have WB images and list human, mouse, and rat reactivity. Choose A03126-2 if your sample is rat brain or mouse Neuro-2a, which appear only in its caption; otherwise compare the reported assay conditions.
Source: BosterBio FUBP1 gene-info card — filtered to Western-blot-capable antibodies; each card shows that product's actual WB validation figure.
References
- UniProt Consortium. UniProt entry Q96AE4.
- Human Protein Atlas. FUBP1 tissue expression.
- PMC5226774 — target-verified WB comparison
- PMC7349734 — target-verified WB comparison
- PMC5447289 — target-verified WB comparison