ELF3 / ETS-related transcription factor Elf-3 · Western blot design guide

Design a Western Blot for ELF3

Real validated ELF3 Western blot protocols, expected-band and isoform facts, troubleshooting for weak or shifted signal, and recommended anti-ELF3 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

Western blot protocol sheet for ELF3: expected band ~41.5 kDa, hero antibody M01027-1, catalog values and labelled standard workflow; separate PMC comparisons on the guide — Printable **ELF3** Western blot protocol sheet — expected band ~41.5 kDa, antibody M01027-1, controls and PMC citations. Open the full ELF3 WB guide →

Navigate to: Protocols Band Controls Antibodies

ELF3 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	~41.5 kDa
	Observed band	~41 kDa
	Gel	10% (catalog M01027-1)
	Positive control ⓘ	Bronchus (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	—
	Caveat	—
	Gene-set association	MSigDB Hallmark membership
	Isoform	2 isoform(s)

Section 1

Real Curated ELF3 Western Blot Protocols

The M01027-1 protocol combines labelled catalog values with standard starting conditions. Published comparisons retain their own sample, reagent and detection scope.

Recommended Western blot protocol parameters
Sample / lysate	human A431, human PANC-1, human A549 (catalog M01027-1)
Gel %	10% (catalog M01027-1)
Load	30 ug; reducing conditions (catalog M01027-1)
Transfer	a nitrocellulose membrane at 150 mA for 50-90 minutes (catalog M01027-1)
Membrane	nitrocellulose membrane (catalog M01027-1)
Blocking	5% non-fat milk/TBS for 1.5 hour at RT (catalog M01027-1)
Primary antibody	M01027-1 · 1:500 (catalog M01027-1)
Primary incubation	overnight at 4°C (catalog M01027-1)
Secondary antibody	goat anti-rabbit IgG-HRP, 1:500 (catalog M01027-1)
Secondary incubation	1.5 hour at RT (catalog M01027-1)
Wash	TBS-0.1%Tween 3 times with 5 minutes each (catalog M01027-1)
Detection	ECL (catalog M01027-1)

Section 2

What Is the Expected ELF3 Western Blot Band Size?

ELF3 is predicted at 41.5 kDa and observed at ~41 kDa; the cause of the small difference is not established.

What am I looking at on my blot?
Band at ~41 kDa	Matches the empirical ELF3 band near its 41.5 kDa predicted mass
Band near 41.5 kDa	Consistent with the predicted ELF3 mass; confirm identity with controls
Doublet near the expected band	Could reflect ELF3 isoforms 1 and 2 if their migration differs; a doublet is not established
Weak ~41 kDa band in one cell fraction	ELF3 occurs in both nucleus and cytoplasm, so fraction abundance may differ

💡Expected ELF3 appearanceELF3 has a predicted mass of 41.5 kDa and an empirical band at ~41 kDa; confirm band identity with suitable positive and negative controls.

How each factor affects band size
Predicted ELF3 mass	Sets a 41.5 kDa reference, close to the empirical ~41 kDa band
Isoform 1	May migrate differently from isoform 2; its individual mass is not supplied
Isoform 2	May migrate differently from isoform 1; its individual mass is not supplied
Alternative splicing	Produces two named isoforms, but distinct band sizes are not established

Why is my band missing or off?
Situation	Likely cause	Next action
No band in lysate	ELF3 may be scarce in the sampled cells	Check a positive control such as A431 lysate and verify sample loading
Band higher than expected	No supplied ELF3 feature establishes a higher band	Check antibody specificity with an ELF3 depletion control
Band lower than expected	No supplied ELF3 feature establishes a lower band	Check antibody specificity with an ELF3 depletion control
Multiple bands	ELF3 has isoforms 1 and 2, though distinct migration is unproven	Test band identity with isoform-specific controls where available
Weak or no signal	ELF3 distribution between nucleus and cytoplasm may affect fraction signal	Compare nuclear and cytoplasmic fractions with a positive control

Source: UniProt P78545 Human Protein Atlas product datasheet (M01027-1)

Experiment evidence

Western blot images showing ELF3 band

M01027-1 · ELF3

A01027-2 · ELF3

← swipe / scroll for more images →

Real WB data behind the band-size expectations in this section.

Sample controls for ELF3 Western blot

🧪HPA-IHC candidate guidance (verify in WB): For positive controls for ELF3 in Western blot, you can use bronchus tissue, which HPA scores High.

Positive control: Bronchus (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: A tissue negative is available, but ELF3's nuclear and cytoplasmic localization may complicate fraction-based controls.

HPA tissue expression evidence for ELF3

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
Bronchus	ciliated cells (cell body)	High	Protein (IHC)	HPA →
Endometrium	glandular cells	High	Protein (IHC)	HPA →
Nasopharynx	ciliated cells (cell body)	High	Protein (IHC)	HPA →
Tonsil	squamous epithelial 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
Adipose tissue	adipocytes	Not detected	Protein (IHC)	HPA →
Adrenal gland	glandular cells	Not detected	Protein (IHC)	HPA →
Bone marrow	hematopoietic cells	Not detected	Protein (IHC)	HPA →
Breast	adipocytes	Not detected	Protein (IHC)	HPA →
Caudate	glial cells	Not detected	Protein (IHC)	HPA →

Section 3

Advanced ELF3 Western Blot Tips

Deeper troubleshooting and optimisation questions for ELF3, answered from its protein features.

How should ELF3 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 ELF3 isoforms produce different bands?

Isoforms · UniProt lists two isoforms. Isoform 2 lacks residues 173–195 in UniProt numbering, so it has a shorter sequence. The supplied features do not establish whether the isoforms resolve as separate bands.

Choose an epitope outside UniProt residues 173–195, which are missing from isoform 2. An antibody against that segment could miss isoform 2. Check the antibody's stated epitope before comparing isoform signals.

Do annotated modifications explain an unexpected ELF3 band shift?

PTM · The supplied UniProt features list no modified residues or glycosylation sites. They therefore provide no specific modification to assign to a shifted band; an unexpected position needs independent verification.

Does this guide establish induction of ELF3?

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 ELF3?

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 M01027-1 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 ELF3 be quantified across cell fractions?

Quantitation · ELF3 is listed in both nucleus and cytoplasm. Quantify each fraction consistently and report which fraction was measured; a change in one fraction need not represent the same change in total ELF3.

How does ELF3's observed band compare with its predicted mass?

Interpretation · The reported band is about 41 kDa, close to the predicted 41.5 kDa. This agreement supports the expected size but does not, by itself, identify the band as ELF3.

Compare them with the expected band near 41 kDa and consider the two listed isoforms. The missing segment in isoform 2 is a possible size distinction, but these features alone cannot identify an extra band.

Boster reagents

ELF3 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

Western blot analysis of ESE1 using anti-ESE1 antibody (M01027-1). Electrophoresis was performed on a 10% SDS-PAGE gel at 80V (Stacking gel) / 120V (Resolving gel) for 2 hours. The sample well of each lane was loaded with 30 ug of sample under reducing conditions. Lane 1: human A431 whole cell lysates, Lane 2: human PANC-1 whole cell lysates, Lane 3: human A549 whole cell lysates, Lane 4: mouse EL-4 whole cell lysates, Lane 5: mouse NIH/3T3 whole cell lysates. After electrophoresis, proteins were transferred to a nitrocellulose membrane at 150 mA for 50-90 minutes. Blocked the membrane with 5% non-fat milk/TBS for 1.5 hour at RT. The membrane was incubated with rabbit anti-ESE1 antigen affinity purified monoclonal antibody (M01027-1) at 1:500 overnight at 4°C, then washed with TBS-0.1%Tween 3 times with 5 minutes each and probed with a goat anti-rabbit IgG-HRP secondary antibody at a dilution of 1:500 for 1.5 hour at RT. The signal is developed using an ECL Plus Western Blotting Substrate (Catalog # AR1196-200) with Tanon 5200 system. A specific band was detected for ESE1 at approximately 41 kDa. The expected band size for ESE1 is at 41 kDa.

Anti-ESE1 Rabbit Monoclonal Antibody

Cat # M01027-1

View product →

Real WB data

Western blot analysis of ELF3 using anti-ELF3 antibody (A01027-2). Electrophoresis was performed on a 10% SDS-PAGE gel at 80V (Stacking gel) / 120V (Resolving gel) for 2 hours. The sample well of each lane was loaded with 30 ug of sample under reducing conditions. Lane 1: human PC-3 whole cell lysates. After electrophoresis, proteins were transferred to a nitrocellulose membrane at 150 mA for 50-90 minutes. Blocked the membrane with 5% non-fat milk/TBS for 1.5 hour at RT. The membrane was incubated with rabbit anti-ELF3 antigen affinity purified polyclonal antibody (A01027-2) at 0.5 μg/mL overnight at 4°C, then washed with TBS-0.1%Tween 3 times with 5 minutes each and probed with a goat anti-rabbit IgG-HRP secondary antibody at a dilution of 1:5000 for 1.5 hour at RT. The signal is developed using an ECL Plus Western Blotting Substrate (Catalog # AR1196-200) with Tanon 5200 system. A specific band was detected for ELF3 at approximately 50 kDa. The expected band size for ELF3 is at 41 kDa.

Anti-ELF3 Antibody Picoband®

Cat # A01027-2

View product →

Two the supplier antibodies for ELF3 have WB images. M01027-1 shows a band near the expected 41 kDa in human and mouse cell lysates. A01027-2 shows a band near 50 kDa in human PC-3 lysate, above its stated 41 kDa expectation.

Which to pick: For human or mouse lysates, M01027-1 has WB examples with a band at the expected size. A01027-2 has a human PC-3 example, but its observed band differs from the expected size. Rat reactivity is listed for M01027-1 without a rat WB example.

Source: BosterBio ELF3 gene-info card — filtered to Western-blot-capable antibodies; each card shows that product's actual WB validation figure.

References

UniProt Consortium. UniProt entry P78545.
Human Protein Atlas. ELF3 tissue expression.
PMC9716751 — target-verified WB comparison
PMC9827823 — target-verified WB comparison
PMC13183993 — target-verified WB comparison