This website uses cookies to ensure you get the best experience on our website.
- Table of Contents
ESRP1 / Epithelial splicing regulatory protein 1 · Western blot design guide
Design a Western Blot for ESRP1
Source-linked ESRP1 Western blot protocol options, expected-band and isoform facts, troubleshooting for weak or shifted signal, and recommended anti-ESRP1 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
ESRP1 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 | ~75.6 kDa | |
| Observed band | ~76 kDa | |
| Gel | 10% (catalog A06068-1) | |
| Positive control | Gallbladder (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
Source-Linked ESRP1 Western Blot Protocol Options
The A06068-1 protocol combines labelled catalog values with standard starting conditions. Published comparisons retain their own sample, reagent and detection scope.
| Sample / lysate | human MCF-7, human RT4 (catalog A06068-1) |
| Gel % | 10% (catalog A06068-1) |
| Load | 30 ug; reducing conditions (catalog A06068-1) |
| Transfer | a nitrocellulose membrane at 150 mA for 50-90 minutes (catalog A06068-1) |
| Membrane | nitrocellulose membrane (catalog A06068-1) |
| Blocking | 5% non-fat milk/TBS for 1.5 hour at RT (catalog A06068-1) |
| Primary antibody | A06068-1 · 1:1000 (catalog A06068-1) |
| Primary incubation | overnight at 4°C (catalog A06068-1) |
| Secondary antibody | goat anti-rabbit IgG-HRP, 1:1000 (catalog A06068-1) |
| Secondary incubation | 1.5 hour at RT (catalog A06068-1) |
| Wash | TBS-0.1%Tween 3 times with 5 minutes each (catalog A06068-1) |
| Detection | ECL (catalog A06068-1) |
Section 2
What Is the Expected ESRP1 Western Blot Band Size?
ESRP1 is predicted at 75.6 kDa and observed at ~76 kDa; the cause of the small difference is not established.
| Band at ~76 kDa | matches the empirical ESRP1 band and its 75.6 kDa predicted mass |
| Closely spaced doublet near ~76 kDa | could include alternative isoforms; their migration is unverified |
| Several discrete bands | could include ESRP1 isoforms 1–5; band identities need confirmation |
| Band enriched in a nuclear fraction | consistent with ESRP1 nuclear localization |
💡Expected ESRP1 appearanceESRP1 is predicted at 75.6 kDa and observed at ~76 kDa in reducing whole-cell blots; confirm band identity with an appropriate positive control and ESRP1 depletion.
| UniProt predicted mass | 75.6 kDa predicted; the empirical ESRP1 band is ~76 kDa |
| Splice isoform 1 | individual mass and migration are not supplied |
| Splice isoform 2 | may differ in size from other isoforms; migration is not supplied |
| Splice isoform 3 | may differ in size from other isoforms; migration is not supplied |
| Splice isoform 4 | may differ in size from other isoforms; migration is not supplied |
| Splice isoform 5 | may differ in size from other isoforms; migration is not supplied |
| Situation | Likely cause | Next action |
|---|---|---|
| No band in lysate | ESRP1 is nuclear and may be scarce in the tested lysate | check a nuclear fraction and an ESRP1-positive cell lysate |
| Band higher than expected | an alternative isoform is possible, but its migration is unknown | compare with the ~76 kDa positive-control band and test ESRP1 depletion |
| Band lower than expected | an alternative isoform is possible, but its migration is unknown | test whether the band decreases after ESRP1 depletion |
| Multiple bands | ESRP1 has five annotated isoforms; individual band assignments are unverified | compare positive controls and test each band's response to ESRP1 depletion |
| Weak or no signal | low ESRP1 abundance in the tested sample is possible | check an ESRP1-positive lysate or enrich the nuclear fraction |
Experiment evidence
Western blot images showing ESRP1 band
A06068-1 · ESRP1
A06068-1 · ESRP1
A06068 · ESRP1
← swipe / scroll for more images →
Real WB data behind the band-size expectations in this section.
Sample controls for ESRP1 Western blot
🧪HPA-IHC candidate guidance (verify in WB): For positive controls for ESRP1 in Western blot, you can use gallbladder tissue, which has high HPA expression.
Positive control: Gallbladder (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 ESRP1.
⚠️Feasibility: ESRP1 is nuclear, so nuclear-enriched lysate may improve detection.
HPA tissue expression evidence for ESRP1
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 |
|---|---|---|---|---|
| Gallbladder | glandular cells | High | Protein (IHC) | HPA → |
| Adrenal gland | glandular cells | Medium | Protein (IHC) | HPA → |
| Appendix | glandular cells | Medium | Protein (IHC) | HPA → |
| Bronchus | respiratory epithelial cells | Medium | Protein (IHC) | HPA → |
| Cerebellum | Purkinje 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 → |
| Liver | cholangiocytes | Not detected | Protein (IHC) | HPA → |
| Salivary gland | glandular cells | Not detected | Protein (IHC) | HPA → |
| Smooth muscle | smooth muscle cells | Not detected | Protein (IHC) | HPA → |
| Soft tissue | chondrocytes | Not detected | Protein (IHC) | HPA → |
Section 3
Advanced ESRP1 Western Blot Tips
Deeper troubleshooting and optimisation questions for ESRP1, answered from its protein features.
How should ESRP1 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 ESRP1 isoforms might produce different bands?
Isoforms · UniProt lists five isoforms. Isoform 2 lacks residues 609–681 and has P608V, so it may migrate differently. Isoforms 3 and 5 lack residues 546–549; isoforms 4 and 5 replace residues 658–681 with CLKDVW. These sequence differences do not establish which bands will be visible.
Check whether the antibody's target region overlaps an altered sequence. Isoforms 3 and 5 lack residues 546–549; isoform 2 lacks 609–681; isoforms 4 and 5 replace 658–681. An antibody targeting one of these regions may detect isoforms differently.
Check whether the antibody's target region overlaps an altered sequence. Isoforms 3 and 5 lack residues 546–549; isoform 2 lacks 609–681; isoforms 4 and 5 replace 658–681. An antibody targeting one of these regions may detect isoforms differently.
Do ESRP1 modifications prove a detectable band shift?
PTM · UniProt lists phosphoserine at position 543 and omega-N-methylarginine at position 582. These are UniProt coordinates for the supplied sequence; other numbering conventions may differ. Their presence alone does not establish a visible shift or explain the observed mass.
Does this guide establish induction of ESRP1?
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 ESRP1?
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 A06068-1 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 ESRP1 bands be quantified across samples?
Quantitation · Quantify the same resolved band across samples and state which band was measured. ESRP1 has five annotated isoforms, so combining bands could obscure differences in isoform abundance. The supplied features do not establish which isoform produces the reported ~76 kDa band.
Why might ESRP1 migrate near 76 kDa?
Interpretation · The predicted mass is 75.6 kDa, close to the reported apparent band at about 76 kDa. The listed features do not establish a cause for any small difference between calculated and apparent mass.
UniProt places ESRP1 in the nucleus. Include a nuclear-containing sample when assessing its presence, and account for fraction choice when comparing band intensity across samples.
Compare their apparent sizes with the ~76 kDa reported band and consider the annotated isoform sequence changes. Check whether the antibody target region is retained in each candidate isoform. The listed phosphorylation and methylation sites alone cannot identify an unexpected band.
UniProt places ESRP1 in the nucleus. Include a nuclear-containing sample when assessing its presence, and account for fraction choice when comparing band intensity across samples.
Compare their apparent sizes with the ~76 kDa reported band and consider the annotated isoform sequence changes. Check whether the antibody target region is retained in each candidate isoform. The listed phosphorylation and methylation sites alone cannot identify an unexpected band.
Boster reagents
ESRP1 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-ESRP1 Antibody
Cat # A06068-1
Real WB data
Anti-RBM35A ESRP1 Antibody
Cat # A06068
Both listed antibodies have WB images. A06068-1 shows a band near 76 kDa in human MCF-7 and RT4 lysates. A06068 is shown with rat colon lysate, with and without blocking peptide. These images document the stated sample contexts only.
Which to pick: For human samples, A06068-1 has illustrated MCF-7 and RT4 results. For rat samples, A06068 lists rat reactivity and has a rat colon image. Both list mouse reactivity, but neither supplied caption shows a mouse sample.
Source: BosterBio ESRP1 gene-info card — filtered to Western-blot-capable antibodies; each card shows that product's actual WB validation figure.
References
- UniProt Consortium. UniProt entry Q6NXG1.
- Human Protein Atlas. ESRP1 tissue expression.
- PMC7906186 — target-verified WB comparison
- PMC5669934 — target-verified WB comparison
- PMC4041859 — target-verified WB comparison