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- Table of Contents
ELF4 / ETS-related transcription factor Elf-4 · Western blot design guide
Design a Western Blot for ELF4
Source-linked ELF4 Western blot protocol options, expected-band and isoform facts, troubleshooting for weak or shifted signal, and recommended anti-ELF4 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
ELF4 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 | ~70.7 kDa | |
| Gel | 12–15% (standard starting point) | |
| Positive control | Adrenal gland (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 | |
| Caveat | Phosphorylation-state controls | |
| Gene-set association | MSigDB Hallmark membership | |
| Isoform | 1 isoform(s) |
Section 1
Source-Linked ELF4 Western Blot Protocol Options
The A03714-2 protocol combines labelled catalog values with standard starting conditions. Published comparisons retain their own sample, reagent and detection scope.
| Sample / lysate | Hela, Raw264.7, H9C2 (catalog A03714-2) |
| 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 | 5% milk or 5% BSA in TBST (standard starting point) |
| Primary antibody | A03714-2; use the WB datasheet starting dilution (standard starting point) |
| Primary incubation | Overnight at 4 °C (standard starting point) |
| Secondary antibody | Species-matched HRP conjugate at validated dilution (standard starting point) |
| Secondary incubation | 1 h at room temperature (standard starting point) |
| Wash | 3 × 5 min in TBST (standard starting point) |
| Detection | ECL; bracket exposures to avoid saturation (standard starting point) |
Section 2
What Is the Expected ELF4 Western Blot Band Size?
ELF4 is predicted at 70.7 kDa; its phosphoserines may affect migration, but no empirical band or visible shift is established.
| Band near 70.7 kDa | Consistent with the predicted ELF4 mass; identity requires confirmation |
| Closely spaced bands near 70.7 kDa | Could reflect ELF4 phosphorylation states; distinct migration is unproven |
| Stronger band in a nuclear fraction | Consistent with ELF4 nuclear localization |
| Weak band in a cytoplasmic fraction | Consistent with ELF4 nuclear localization |
💡Expected ELF4 appearanceELF4 has a predicted mass of 70.7 kDa, but no empirical band size is supplied; confirm any candidate band with ordinary identity controls.
| Predicted ELF4 mass | Sets a 70.7 kDa reference, not a measured migration position |
| Phosphoserine 151 | May affect mobility; no shift is established |
| Phosphoserine 188 | May affect mobility; no shift is established |
| Phosphoserine 641 | May affect mobility; no shift is established |
| Phosphoserine 648 | May affect mobility; no shift is established |
| Situation | Likely cause | Next action |
|---|---|---|
| No band in lysate | Nuclear ELF4 may be poorly recovered | Check nuclear extraction and use a validated positive lysate |
| Band higher than expected | Phosphorylation may affect migration, but the cause is unproven | Compare phosphatase-treated material and confirm band identity |
| Band lower than expected | The lower band has no established explanation in the supplied features | Confirm specificity with ELF4 depletion or an independent antibody |
| Multiple bands | Phosphorylation states are possible; distinct bands are unproven | Compare phosphatase treatment and ELF4 depletion |
| Weak or no signal | A whole-cell preparation may underrepresent nuclear ELF4 | Check a nuclear fraction and verify loading |
Experiment evidence
Western blot images showing ELF4 band
A03714-2 · ELF4
A03714-1 · ELF4
A03714-1 · ELF4
← swipe / scroll for more images →
Real WB data behind the band-size expectations in this section.
Sample controls for ELF4 Western blot
🧪HPA-IHC candidate guidance (verify in WB): For positive controls for ELF4 in Western blot, you can use adrenal gland lysate.
Positive control: Adrenal gland (IHC candidate; verify WB)
Negative control: Adipose tissue (IHC candidate; verify WB)
Loading controls: Run GAPDH, β-actin, and a total-protein stain alongside.
⚠️Feasibility: ELF4 is nuclear, so nuclear-enriched lysate may improve detection.
HPA tissue expression evidence for ELF4
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 |
|---|---|---|---|---|
| Adrenal gland | glandular cells | Medium | Protein (IHC) | HPA → |
| Appendix | glandular cells | Medium | Protein (IHC) | HPA → |
| Bronchus | respiratory epithelial cells | Medium | Protein (IHC) | HPA → |
| Cervix | squamous epithelial cells | Medium | Protein (IHC) | HPA → |
| Colon | glandular 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 → |
| Caudate | glial cells | Not detected | Protein (IHC) | HPA → |
| Cerebellum | cells in granular layer | Not detected | Protein (IHC) | HPA → |
| Cerebral cortex | endothelial cells | Not detected | Protein (IHC) | HPA → |
| Heart muscle | cardiomyocytes | Not detected | Protein (IHC) | HPA → |
Section 3
Advanced ELF4 Western Blot Tips
Deeper troubleshooting and optimisation questions for ELF4, answered from its protein features.
What band size is predicted for ELF4?
Band shift · The 663-residue ELF4 sequence has a predicted mass of 70.7 kDa. No empirical band position is supplied, so use 70.7 kDa as a reference, not a confirmed migration position.
Do annotated isoforms explain multiple ELF4 bands?
Isoforms · Only one ELF4 isoform is listed, with no alternative sequence supplied. Do not assign additional bands to annotated isoforms on this evidence alone.
Could ELF4 phosphorylation affect its migration?
PTM · UniProt lists phosphoserine at positions 151, 188, 641, and 648, using UniProt sequence numbering. Compare untreated and phosphatase-treated samples if you suspect phosphorylation affects migration. These sites alone do not establish a visible shift.
Does this guide establish induction of ELF4?
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 ELF4?
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 A03714-2 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 ELF4 be quantified across samples?
Quantitation · Quantify ELF4 in matched sample fractions and account for their loading. Because ELF4 localizes to the nucleus and PML nuclear bodies, a change in one fraction does not by itself establish a change in total ELF4.
Which fraction should be examined for ELF4?
Interpretation · ELF4 is annotated in the nucleus and PML nuclear bodies. A nuclear fraction is appropriate for assessing its signal; compare equivalent fractions across samples.
ELF4 interacts with PML through its C-terminus, and PML mediates its accumulation in PML nuclear bodies. Changes in that localization could alter a compartment-specific signal, so interpret fraction measurements alongside localization.
Confirm band identity before assigning a cause. The supplied features list phosphoserines but do not establish a migration shift; they list no alternative sequence, signal peptide, propeptide, or glycosylation sites. No empirical ELF4 band position is available for comparison.
ELF4 interacts with PML through its C-terminus, and PML mediates its accumulation in PML nuclear bodies. Changes in that localization could alter a compartment-specific signal, so interpret fraction measurements alongside localization.
Confirm band identity before assigning a cause. The supplied features list phosphoserines but do not establish a migration shift; they list no alternative sequence, signal peptide, propeptide, or glycosylation sites. No empirical ELF4 band position is available for comparison.
Boster reagents
ELF4 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-Elf-4 Antibody
Cat # A03714-2
Real WB data
Anti-Elf-4 Antibody
Cat # A03714-1
Two the supplier anti-ELF4 antibodies have WB images. A03714-2 was shown at 1:500 with HeLa, RAW264.7, and H9C2 lysates; A03714-1 was shown with RAW264.7 lysate. The supplied evidence does not establish broader performance across all listed species.
Which to pick: Choose A03714-2 for its listed human, mouse, and rat reactivity and WB image using three named lysates. A03714-1 lists human and mouse reactivity, with a WB image using RAW264.7 lysate. Match your sample to the reported context.
Source: BosterBio ELF4 gene-info card — filtered to Western-blot-capable antibodies; each card shows that product's actual WB validation figure.
References
- UniProt Consortium. UniProt entry Q99607.
- Human Protein Atlas. ELF4 tissue expression.
- PMC12645360 — target-verified WB comparison
- PMC10875363 — target-verified WB comparison
- PMC10360327 — target-verified WB comparison