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- Table of Contents
ELF1 / ETS-related transcription factor Elf-1 · Western blot design guide
Design a Western Blot for ELF1
Real validated ELF1 Western blot protocols, expected-band and isoform facts, troubleshooting for weak or shifted signal, and recommended anti-ELF1 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
ELF1 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 | ~67.5 kDa | |
| Observed band | ~75 kDa | |
| Gel | 5–20% (catalog A03187-1) | |
| Positive control | Lymph node (IHC candidate; verify WB) +4 more | |
| Negative control | Cerebellum (IHC candidate; verify WB) |
Important caveats
Reasons your observed band may differ from the expected size.
| PTM | Phosphorylated | |
| Caveat | Phosphorylation-state controls | |
| Gene-set association | MSigDB Hallmark membership | |
| Isoform | 2 isoform(s) |
Section 1
Real Curated ELF1 Western Blot Protocols
The A03187-1 protocol combines labelled catalog values with standard starting conditions. Published comparisons retain their own sample, reagent and detection scope.
| Sample / lysate | human Hela, human COLO-320, human A549 (catalog A03187-1) |
| Gel % | 5–20% (catalog A03187-1) |
| Load | 50ug; reducing conditions (catalog A03187-1) |
| Transfer | a Nitrocellulose membrane at 150mA for 50-90 minutes (catalog A03187-1) |
| Membrane | Nitrocellulose membrane (catalog A03187-1) |
| Blocking | 5% Non-fat Milk/ TBS for 1.5 hour at RT (catalog A03187-1) |
| Primary antibody | A03187-1 · 0.5 μg/mL (catalog A03187-1) |
| Primary incubation | overnight at 4°C (catalog A03187-1) |
| Secondary antibody | goat anti-rabbit IgG-HRP, 1:10000 (catalog A03187-1) |
| Secondary incubation | 1.5 hour at RT (catalog A03187-1) |
| Wash | TBS-0.1%Tween 3 times with 5 minutes each (catalog A03187-1) |
| Detection | ECL (catalog A03187-1) |
Section 2
What Is the Expected ELF1 Western Blot Band Size?
ELF1 is predicted at 67.5 kDa and observed near 75 kDa; the cause of the difference is not established.
| Band near 75 kDa | Empirical ELF1 band in reducing whole-cell blots; confirm identity with an independent antibody or ELF1 depletion. |
| Band near 67.5 kDa | Near the predicted full-length mass; identity still requires confirmation. |
| Multiple bands at different positions | Isoforms 1 and 2 or different phosphorylation states are possible; distinct migration is unproven. |
| Closely spaced doublet | Different phosphorylation states are possible; the listed sites do not establish a visible shift. |
💡Expected ELF1 appearanceELF1 has a predicted mass of 67.5 kDa and an empirical band near 75 kDa in reducing whole-cell blots; the cause of the difference is unestablished, so confirm identity with an independent antibody or ELF1 depletion.
| Predicted ELF1 mass | 67.5 kDa by sequence; the empirical band is near 75 kDa, with no established cause for the difference. |
| Isoform 1 | May differ in size from isoform 2; its mass and migration are not supplied. |
| Isoform 2 | May differ in size from isoform 1; its mass and migration are not supplied. |
| Listed phosphoserine and phosphothreonine sites | Phosphorylation could affect migration, but no visible shift or contribution to the 75 kDa band is established. |
| Situation | Likely cause | Next action |
|---|---|---|
| No band in lysate | Nuclear ELF1 may be poorly recovered during lysate preparation. | Check nuclear protein recovery and compare with a nuclear-enriched fraction. |
| Band higher than expected | The observed ELF1 band is near 75 kDa rather than the predicted 67.5 kDa; its cause is unestablished. | Confirm identity with an independent antibody or ELF1 depletion. |
| Band lower than expected | An isoform or protein breakdown is possible; no smaller ELF1 mass is supplied. | Use ELF1 depletion and a second antibody to identify the band. |
| Multiple bands | Isoforms 1 and 2 or different phosphorylation states are possible, but distinct migration is unproven. | Compare bands after ELF1 depletion; test phosphatase sensitivity if needed. |
| Weak or no signal | Nuclear ELF1 may be underrepresented in the sample. | Check sample recovery and enrich the nuclear fraction. |
| Fragments below expected size | Protein breakdown is possible; no ELF1 fragment sizes are supplied. | Prepare fresh lysate with protease inhibitors and confirm band identity. |
Experiment evidence
Western blot images showing ELF1 band
A03187-1 · ELF1
← swipe / scroll for more images →
Real WB data behind the band-size expectations in this section.
Sample controls for ELF1 Western blot
🧪HPA-IHC candidate guidance (verify in WB): For positive controls for ELF1 in Western blot, you can use lymph node tissue, which HPA rates High for ELF1.
Positive control: Lymph node (IHC candidate; verify WB)
Negative control: Cerebellum (IHC candidate; verify WB)
Loading controls: Run GAPDH, β-actin, and a total-protein stain alongside the samples.
⚠️Feasibility: ELF1 is nuclear, so a nuclear-enriched lysate may improve detection.
HPA tissue expression evidence for ELF1
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 |
|---|---|---|---|---|
| Lymph node | germinal center cells | High | Protein (IHC) | HPA → |
| Nasopharynx | respiratory epithelial cells | High | Protein (IHC) | HPA → |
| Oral mucosa | squamous epithelial cells | High | Protein (IHC) | HPA → |
| Tonsil | germinal center cells | High | Protein (IHC) | HPA → |
| Urinary bladder | urothelial cells | High | Protein (IHC) | HPA → |
Lower expression tissues · IHC evidence, not confirmed WB-negative controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| Cerebellum | cells in granular layer | Not detected | Protein (IHC) | HPA → |
| Heart muscle | cardiomyocytes | Not detected | Protein (IHC) | HPA → |
| Skeletal muscle | myocytes | Not detected | Protein (IHC) | HPA → |
| Smooth muscle | smooth muscle cells | Not detected | Protein (IHC) | HPA → |
| Adipose tissue | adipocytes | Low | Protein (IHC) | HPA → |
Section 3
Advanced ELF1 Western Blot Tips
Deeper troubleshooting and optimisation questions for ELF1, answered from its protein features.
How should ELF1 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 ELF1 isoforms produce more than one band?
Isoforms · UniProt lists isoforms 1 and 2. Isoform 2 lacks residues 122–145 of the canonical sequence, so a second band is plausible. The feature does not establish whether the isoforms separate visibly or how strongly either is expressed.
Which phosphorylation sites matter when interpreting ELF1 bands?
PTM · UniProt annotates phosphoserine at canonical positions 110, 163, 167, 168, 187, and 432, and phosphothreonine at 190. These are UniProt coordinates; antibody or paper numbering may differ. Their annotation alone does not demonstrate a visible band shift.
Does this guide establish induction of ELF1?
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 ELF1 Western blot?
Transfer · ELF1 is a 619-residue protein with a predicted mass of 67.5 kDa and an observed band near 75 kDa. Choose transfer conditions suitable for that size range and check transfer efficiency around 75 kDa. The supplied features do not establish a preferred transfer method.
How should blocking be optimized?
Blocking · Standard workflow guidance: follow the A03187-1 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 ELF1 band intensity be quantified?
Quantitation · Measure the same identified ELF1 band across samples and use a consistent normalization control. If multiple bands appear, identify them before combining their signals: UniProt lists two isoforms and several phosphorylation sites, but does not establish which bands correspond to either.
Why might ELF1 appear near 75 kDa instead of 67.5 kDa?
Interpretation · The supplied observed band is about 75 kDa, while the predicted mass is 67.5 kDa. ELF1 has annotated phosphorylation sites, but their presence alone does not explain this difference. Confirm band identity with an appropriate ELF1 control before assigning the band.
UniProt places ELF1 in the nucleus. A nuclear fraction can enrich the target when whole-cell signal is weak; check fraction quality when comparing samples.
Compare bands with the expected 67.5 kDa mass and the reported apparent band near 75 kDa. Isoform 2 lacks canonical residues 122–145, and phosphorylation is annotated at seven sites. Neither feature by itself identifies an unexpected band; verify its identity before assigning it to an isoform or modification.
UniProt places ELF1 in the nucleus. A nuclear fraction can enrich the target when whole-cell signal is weak; check fraction quality when comparing samples.
Compare bands with the expected 67.5 kDa mass and the reported apparent band near 75 kDa. Isoform 2 lacks canonical residues 122–145, and phosphorylation is annotated at seven sites. Neither feature by itself identifies an unexpected band; verify its identity before assigning it to an isoform or modification.
Boster reagents
ELF1 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-E74 like factor 1/ELF1 Antibody Picoband®
Cat # A03187-1
The catalog reports A03187-1, a rabbit polyclonal anti-ELF1 antibody with reported human, mouse, and rat reactivity. Its WB image uses seven human cell lysates and shows a band near 75 kDa, compared with an expected 67 kDa; the supplied image does not test mouse or rat samples.
Which to pick: A03187-1 is the only listed option and has a WB image from human cell lysates. For mouse or rat samples, the catalog reports reactivity, but the supplied WB image provides no example in those species.
Source: BosterBio ELF1 gene-info card — filtered to Western-blot-capable antibodies; each card shows that product's actual WB validation figure.
References
- UniProt Consortium. UniProt entry P32519.
- Human Protein Atlas. ELF1 tissue expression.
- PMC11535329 — target-verified WB comparison
- PMC13483860 — target-verified WB comparison
- PMC7294115 — target-verified WB comparison