This website uses cookies to ensure you get the best experience on our website.
- Table of Contents
ETS2 / Protein C-ets-2 · Western blot design guide
Design a Western Blot for ETS2
Real validated ETS2 Western blot protocols, expected-band and isoform facts, troubleshooting for weak or shifted signal, and recommended anti-ETS2 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
ETS2 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 | ~53 kDa | |
| Gel | 12–15% (standard starting point) | |
| Negative control | Suggested KO / knockdown lysate |
Important caveats
Reasons your observed band may differ from the expected size.
| PTM | Phosphorylated | |
| Caveat | SPRY3 antibody mismatch | |
| Gene-set association | MSigDB Hallmark membership | |
| Isoform | 1 isoform(s) |
Section 1
Real Curated ETS2 Western Blot Protocols
The A02641 protocol combines labelled catalog values with standard starting conditions. Published comparisons retain their own sample, reagent and detection scope.
| Sample / lysate | extracts of various cell lines, (catalog A02641) |
| Gel % | 12–15% (standard starting point) |
| Load | 20–30 µg total protein per lane; optimize for abundance (standard starting point) |
| Transfer | Standard semi-dry transfer; verify efficiency (standard starting point) |
| Membrane | 0.45 µm PVDF (standard starting point) |
| Blocking | Blocking buffer: 3% nonfat dry milk in TBST (catalog A02641) |
| Primary antibody | A02641 · 1:1000 (catalog A02641) |
| Primary incubation | Overnight at 4 °C (standard starting point) |
| Secondary antibody | Goat Anti-Rabbit IgG, 1:10000 (catalog A02641) |
| Secondary incubation | 1 h at room temperature (standard starting point) |
| Wash | 3 × 5 min in TBST (standard starting point) |
| Detection | ECL (catalog A02641) |
Section 2
What Is the Expected ETS2 Western Blot Band Size?
ETS2 is predicted at 53 kDa; its phosphoserines could affect mobility, but no migration shift or empirical band is demonstrated.
| Band near 53 kDa | Consistent with full-length ETS2, pending identity controls |
| Slightly slower band near 53 kDa | Could reflect phosphorylation, but a mobility shift is unproven |
| Close doublet near 53 kDa | Could reflect different phosphorylation states, pending phosphatase testing |
| Stronger band in nuclear extract | Consistent with the reported nuclear location |
💡Expected ETS2 appearanceETS2 has a predicted mass of 53 kDa and no supplied empirical band; assess a band near 53 kDa with antibody and sample controls because the listed phosphoserines do not establish its migration.
| Predicted molecular mass | Full-length ETS2 is predicted at 53 kDa |
| Phosphoserines 220 and 225 | May affect mobility; no shift is established |
| Phosphoserines 295 and 298 | May affect mobility; no shift is established |
| Phosphoserine 301 | May affect mobility; no shift is established |
| Situation | Likely cause | Next action |
|---|---|---|
| No band in lysate | Nuclear ETS2 may be underrepresented in the sample | Check nuclear enrichment and a positive-control lysate |
| Band higher than expected | Phosphorylation could affect mobility, but the cause is unestablished | Compare phosphatase-treated and untreated samples and verify band identity |
| Band lower than expected | Possible degradation; no cleavage feature is listed | Check sample handling and confirm with an independent ETS2 antibody |
| Multiple bands | Different phosphorylation states are possible but unproven | Compare phosphatase-treated samples and use an independent ETS2 antibody |
| Weak or no signal | The nuclear target may be insufficiently represented | Enrich the nuclear fraction and check the antibody with a positive control |
| Fragments below expected size | Possible sample proteolysis | Use fresh lysate with protease inhibitors and confirm fragment identity |
Experiment evidence
Western blot images showing ETS2 band
A02641 · ETS2
← swipe / scroll for more images →
Real WB data behind the band-size expectations in this section.
Sample controls for ETS2 Western blot
🧪For positive controls for ETS2 in Western blot, you can use a validated ETS2-expressing sample, but the supplied HPA evidence identifies no positive tissue or cell line.
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 such as Ponceau alongside the samples.
⚠️Feasibility: HPA expression data are unavailable, so tissue controls cannot be selected from the supplied evidence.
HPA tissue expression evidence for ETS2
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 ETS2 Western Blot Tips
Deeper troubleshooting and optimisation questions for ETS2, answered from its protein features.
Where should the ETS2 band appear?
Band shift · The predicted mass is 53 kDa. No observed band position is supplied, so use 53 kDa as a reference rather than an established apparent mass.
Could an ETS2 isoform explain a second band?
Isoforms · Only one isoform is listed, with no alternative sequence supplied. The provided features therefore do not support assigning a second band to a known ETS2 isoform.
Can phosphorylation explain a shifted ETS2 band?
PTM · ETS2 has five listed phosphoserines, but their presence alone does not establish a visible shift or explain a difference from 53 kDa. Compare samples under matched conditions before attributing a shifted band to phosphorylation.
The supplied UniProt positions are Ser220, Ser225, Ser295, Ser298, and Ser301. Use these coordinates when checking antibody specificity; numbering in a paper or antibody description may follow a different convention.
The supplied UniProt positions are Ser220, Ser225, Ser295, Ser298, and Ser301. Use these coordinates when checking antibody specificity; numbering in a paper or antibody description may follow a different convention.
Does this guide establish induction of ETS2?
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 ETS2?
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 A02641 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.
Which fraction is appropriate for ETS2 quantitation?
Quantitation · ETS2 is listed in the nucleus. Use comparable nuclear samples when quantifying its band, and keep the sample fraction consistent across conditions.
How should unexpected ETS2 bands be interpreted?
Interpretation · Start with the 53 kDa prediction and the listed nuclear location. The supplied features do not identify an observed band, cleavage product, glycosylation site, or alternative isoform. Do not assign another band to ETS2 or a specific modification from size alone.
Boster reagents
ETS2 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-Protein C-ets-2 ETS2 Antibody
Cat # A02641
A02641 is the listed anti-ETS2 antibody, with reported human, mouse and rat reactivity. Its WB image caption describes various cell lines but names SPRY3 antibody, so the image does not clearly establish ETS2 WB validation.
Which to pick: Only A02641 is listed. It has a WB image, but the SPRY3 caption makes that evidence ambiguous; confirm the antibody identity and suitability for your samples before use.
Source: BosterBio ETS2 gene-info card — filtered to Western-blot-capable antibodies; each card shows that product's actual WB validation figure.
References
- UniProt Consortium. UniProt entry P15036.
- Human Protein Atlas. ETS2 tissue expression.
- PMC4072627 — target-verified WB comparison
- PMC9822945 — target-verified WB comparison
- PMC3846434 — target-verified WB comparison