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- Table of Contents
ETS1 / Protein C-ets-1 · Western blot design guide
Design a Western Blot for ETS1
Real validated ETS1 Western blot protocols, expected-band and isoform facts, troubleshooting for weak or shifted signal, and recommended anti-ETS1 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
ETS1 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 | ~50.4 kDa | |
| Observed band | ~50 kDa | |
| Gel | 10% (catalog A00931-2) | |
| Positive control | Appendix (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 | Modification-state controls | |
| Gene-set association | MSigDB Hallmark membership | |
| Isoform | 5 isoform(s) |
Section 1
Real Curated ETS1 Western Blot Protocols
The A00931-2 protocol combines labelled catalog values with standard starting conditions. Published comparisons retain their own sample, reagent and detection scope.
| Sample / lysate | human Jurkat, human Daudi, mouse thymus (catalog A00931-2) |
| Gel % | 10% (catalog A00931-2) |
| Load | 30 ug; reducing conditions (catalog A00931-2) |
| Transfer | a nitrocellulose membrane at 150 mA for 50-90 minutes (catalog A00931-2) |
| Membrane | nitrocellulose membrane (catalog A00931-2) |
| Blocking | 5% non-fat milk/TBS for 1.5 hour at RT (catalog A00931-2) |
| Primary antibody | A00931-2 · 0.5 μg/mL (catalog A00931-2) |
| Primary incubation | overnight at 4°C (catalog A00931-2) |
| Secondary antibody | goat anti-rabbit IgG-HRP, 1:5000 (catalog A00931-2) |
| Secondary incubation | 1.5 hour at RT (catalog A00931-2) |
| Wash | TBS-0.1%Tween 3 times with 5 minutes each (catalog A00931-2) |
| Detection | ECL (catalog A00931-2) |
Section 2
What Is the Expected ETS1 Western Blot Band Size?
ETS1 is predicted at 50.4 kDa and observed at approximately 50 kDa; the small difference has no established cause in the supplied evidence.
| Band near 50 kDa | Observed ETS1 band, consistent with the 50.4 kDa predicted mass |
| Additional bands at different positions | Could reflect named ETS1 isoforms; their migration is not supplied |
| Band near twice the main band size | Could reflect a homodimer retained during electrophoresis; its SDS stability is unproven |
| Slight shift or doublet near 50 kDa | Could reflect modified ETS1; a visible shift is not established |
💡Expected ETS1 appearanceThe predicted ETS1 mass is 50.4 kDa, and an approximately 50 kDa band was observed in reducing whole-cell and tissue blots; confirm identity with a suitable positive control and antibody-specificity checks.
| Predicted ETS1 mass | Places the full-length sequence near 50.4 kDa, consistent with the observed approximately 50 kDa band |
| Isoforms c-ETS-1A, c-ETS-1B, 3, Ets-1 p27, and 5 | May differ in size, but distinct band positions are not established |
| DNA-binding homodimer | Could appear near twice the monomer size if it survives electrophoresis |
| Phosphorylation at Thr38, Tyr223, Ser251, Ser254, Thr265, and Ser267 | Could alter migration, but a visible shift is not established |
| Alternate acetylation at Lys8 and Lys15 | Could alter migration, but a visible shift is not established |
| Situation | Likely cause | Next action |
|---|---|---|
| No band in lysate | ETS1 may be below detection in the tested sample | Compare with a documented positive lysate and check loading and transfer |
| Band higher than expected | A retained homodimer or modified ETS1 is possible, but neither explains a shift without validation | Compare reducing and non-reducing preparations and verify band identity |
| Band lower than expected | An ETS1 splice isoform is possible; its band size is not supplied | Check antibody epitope coverage and verify the band with an independent antibody |
| Multiple bands | ETS1 has five named isoforms and several modification sites, but their band patterns are unestablished | Compare a positive lysate and test band identity with an independent antibody |
| Weak or no signal | ETS1 distribution between nucleus and cytoplasm may affect fraction signals | Check both fractions and compare with a documented positive lysate |
Experiment evidence
Western blot images showing ETS1 band
A00931-2 · ETS1
M00931-1 · ETS1
← swipe / scroll for more images →
Real WB data behind the band-size expectations in this section.
Sample controls for ETS1 Western blot
🧪HPA-IHC candidate guidance (verify in WB): For positive controls for ETS1 in Western blot, you can use appendix tissue lysate, which HPA scores High for ETS1.
Positive control: Appendix (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: Tissue controls are feasible, though ETS1 can localize to both the nucleus and cytoplasm.
HPA tissue expression evidence for ETS1
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 |
|---|---|---|---|---|
| Appendix | lymphoid tissue | High | Protein (IHC) | HPA → |
| Bone marrow | hematopoietic cells | High | Protein (IHC) | HPA → |
| Lymph node | non-germinal center cells | High | Protein (IHC) | HPA → |
| Tonsil | germinal center cells | High | Protein (IHC) | HPA → |
| Spleen | cells in red pulp | 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 → |
| Adrenal gland | glandular cells | Not detected | Protein (IHC) | HPA → |
| Breast | adipocytes | Not detected | Protein (IHC) | HPA → |
| Bronchus | respiratory epithelial cells | Not detected | Protein (IHC) | HPA → |
| Caudate | glial cells | Not detected | Protein (IHC) | HPA → |
Section 3
Advanced ETS1 Western Blot Tips
Deeper troubleshooting and optimisation questions for ETS1, answered from its protein features.
How should ETS1 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 ETS1 isoforms could produce different bands?
Isoforms · UniProt lists five isoforms. Relative to the 441-residue canonical sequence, Ets-1 p27 lacks residues 28–243, c-ETS-1B lacks 244–330, and isoform 5 replaces 262–272 and lacks 273–441. Isoform 3 has a different N-terminal sequence at 1–27. Consider these sequence differences when assigning additional bands.
Check the antibody epitope against the listed sequence changes. An epitope within canonical residues 28–243 is absent from Ets-1 p27; one within 244–330 is absent from c-ETS-1B. Isoform 5 lacks 273–441, and isoform 3 replaces 1–27. An undetected isoform may therefore reflect its sequence rather than its abundance.
Check the antibody epitope against the listed sequence changes. An epitope within canonical residues 28–243 is absent from Ets-1 p27; one within 244–330 is absent from c-ETS-1B. Isoform 5 lacks 273–441, and isoform 3 replaces 1–27. An undetected isoform may therefore reflect its sequence rather than its abundance.
Which ETS1 modifications are relevant to band interpretation?
PTM · At UniProt canonical coordinates, phosphorylation is listed at Thr38 by MAPK, Tyr223, Ser251, Ser254, Thr265, Ser267, Ser270, Ser282 and Ser285; acetylation is listed at Lys8, Lys15 and Lys305. These features identify possible forms but do not establish that they resolve as separate bands.
UniProt assigns MAPK-dependent phosphorylation to Thr38. Compare total ETS1 with a Thr38-specific signal, if available, across matched conditions that alter MAPK activity. The supplied features specify neither an induction condition nor a visible band shift.
UniProt assigns MAPK-dependent phosphorylation to Thr38. Compare total ETS1 with a Thr38-specific signal, if available, across matched conditions that alter MAPK activity. The supplied features specify neither an induction condition nor a visible band shift.
Does this guide establish induction of ETS1?
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 ETS1?
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 A00931-2 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 should nuclear and cytoplasmic ETS1 be quantified?
Quantitation · Quantify the compartments separately if localization matters. ETS1 is listed in both nucleus and cytoplasm, and coexpression with Ets-1 p27 delocalizes ETS1 from nucleus to cytoplasm. A change in one fraction need not represent a change in total ETS1.
How does the observed ETS1 band compare with its predicted mass?
Interpretation · The observed band is approximately 50 kDa, close to the canonical predicted mass of 50.4 kDa. The listed modifications do not, by themselves, establish a visible shift or explain any small difference in apparent mass.
Compare its detection with the antibody epitope and the listed isoforms before assigning it. Ets-1 p27 lacks canonical residues 28–243, c-ETS-1B lacks 244–330, and isoform 5 lacks 273–441. The supplied features alone cannot identify a particular observed band.
Compare its detection with the antibody epitope and the listed isoforms before assigning it. Ets-1 p27 lacks canonical residues 28–243, c-ETS-1B lacks 244–330, and isoform 5 lacks 273–441. The supplied features alone cannot identify a particular observed band.
Boster reagents
ETS1 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-ETS1 Antibody Picoband®
Cat # A00931-2
Real WB data
Anti-ETS1 Monoclonal Antibody
Cat # M00931-1
Both the supplier anti-ETS1 antibodies list human and mouse reactivity and have Western blot images showing an approximately 50 kDa band. A00931-2 was tested on human cell and mouse thymus lysates; M00931-1 was imaged with human cell lysates only.
Which to pick: For mouse tissue, A00931-2 has a WB image using mouse thymus lysate. For human cell lysates, both have WB images: A00931-2 with Jurkat and Daudi; M00931-1 with Jurkat, HEL, RT4, and U251. The listed mouse reactivity of M00931-1 lacks an accompanying mouse WB image.
Source: BosterBio ETS1 gene-info card — filtered to Western-blot-capable antibodies; each card shows that product's actual WB validation figure.
References
- UniProt Consortium. UniProt entry P14921.
- Human Protein Atlas. ETS1 tissue expression.
- PMC10444662 — target-verified WB comparison
- PMC5448599 — target-verified WB comparison
- PMC4045976 — target-verified WB comparison