TEAD1 / Transcriptional enhancer factor TEF-1 · Western blot design guide

Design a Western Blot for TEAD1

Real validated TEAD1 Western blot protocols, expected-band and isoform facts, troubleshooting for weak or shifted signal, and recommended anti-TEAD1 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 TEAD1: expected band ~47.9 kDa, hero antibody M03263-1, catalog values and labelled standard workflow; separate PMC comparisons on the guide — Printable **TEAD1** Western blot protocol sheet — expected band ~47.9 kDa, antibody M03263-1, controls and PMC citations. Open the full TEAD1 WB guide →

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TEAD1 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	~47.9 kDa
	Observed band	~52 kDa
	Gel	5–20% (catalog M03263-1)
	Negative control ⓘ	Suggested KO / knockdown lysate

Important caveats

Reasons your observed band may differ from the expected size.

	PTM	Phosphorylated + Acetylated
	Caveat	Phosphorylation-state controls
	Gene-set association	MSigDB C7 membership
	Isoform	2 isoform(s)

Section 1

Real Curated TEAD1 Western Blot Protocols

The M03263-1 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 Hela, human SiHa (catalog M03263-1)
Gel %	5–20% (catalog M03263-1)
Load	30 ug; reducing conditions (catalog M03263-1)
Transfer	a nitrocellulose membrane at 150 mA for 50-90 minutes (catalog M03263-1)
Membrane	nitrocellulose membrane (catalog M03263-1)
Blocking	5% non-fat milk/TBS for 1.5 hour at RT (catalog M03263-1)
Primary antibody	M03263-1 · 1:1000 (catalog M03263-1)
Primary incubation	overnight at 4°C (catalog M03263-1)
Secondary antibody	goat anti-rabbit IgG-HRP, 1:500 (catalog M03263-1)
Secondary incubation	1.5 hour at RT (catalog M03263-1)
Wash	TBS-0.1%Tween 3 times with 5 minutes each (catalog M03263-1)
Detection	ECL (catalog M03263-1)

Section 2

What Is the Expected TEAD1 Western Blot Band Size?

TEAD1 is predicted at 47.9 kDa and observed near 52 kDa; the cause of the difference is not established.

What am I looking at on my blot?
Band near 52 kDa	Empirical TEAD1 band in reducing whole-cell lysates; confirm identity with controls
Band near 47.9 kDa	Near the predicted TEAD1 mass; identity requires validation
Multiple bands	TEAD1 has isoforms 1 and 2, but distinct migration is unverified
Faint band in whole-cell lysate	Nuclear TEAD1 may be more detectable after nuclear enrichment

💡Expected TEAD1 appearanceTEAD1 has a predicted mass of 47.9 kDa and an empirical band near 52 kDa in reducing whole-cell lysates; use band-identity controls because the cause of the difference is unestablished.

How each factor affects band size
Predicted TEAD1 mass	47.9 kDa predicted; the empirical band is near 52 kDa
Isoforms 1 and 2	May differ in size, but their masses and migration are not supplied
Phosphoserine at residue 11	May affect mobility; no visible shift is established
N6-lactoyllysine at residue 108	May affect mobility; no visible shift is established
N-acetylmethionine at residue 1	Its effect on apparent band size is not established

Why is my band missing or off?
Situation	Likely cause	Next action
No band in lysate	Nuclear TEAD1 may be diluted in whole-cell lysate	Check a nuclear fraction and a positive-control lysate
Band higher than expected	The empirical TEAD1 band is near 52 kDa, above its 47.9 kDa predicted mass; the cause is unestablished	Compare with the reported band and verify identity by TEAD1 knockdown
Band lower than expected	Isoform 2 may differ in size, but its migration is unknown	Check band identity by TEAD1 knockdown before assigning an isoform
Multiple bands	TEAD1 has two isoforms, but distinct bands are unverified	Use TEAD1 knockdown to identify specific bands
Weak or no signal	Nuclear TEAD1 may be underrepresented in the tested sample	Enrich nuclei and include a positive-control lysate

Source: UniProt P28347 Human Protein Atlas product datasheet (M03263-1)

Experiment evidence

Western blot images showing TEAD1 band

M03263-1 · TEAD1

M03263-2 · TEAD1

A03263-2 · TEAD1

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

Sample controls for TEAD1 Western blot

🧪For positive controls for TEAD1 in Western blot, you can use a sample with independently confirmed TEAD1 expression; the supplied HPA evidence identifies no positive tissue or cell.

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 alongside the samples.

⚠️Feasibility: With no HPA expression data, a positive sample needs validation and siRNA knockdown or a KO line can provide a negative control.

HPA tissue expression evidence for TEAD1

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 TEAD1 Western Blot Tips

Deeper troubleshooting and optimisation questions for TEAD1, answered from its protein features.

How should TEAD1 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 TEAD1 isoforms produce different bands?

Isoforms · UniProt lists two isoforms. Relative to isoform 1, isoform 2 lacks residues 1–15 and 234–291, and replaces K110 with KVTSM. These sequence changes could affect band size, but the supplied features do not establish where either isoform migrates. Check which isoforms the antibody recognizes.

Isoform 2 lacks canonical residues 1–15, including phosphoserine 11. A signal from an antibody specific to canonical phosphoserine 11 therefore should not be assigned to isoform 2. Verify antibody specificity and use canonical UniProt numbering when reporting the site.

Which TEAD1 modifications matter when interpreting bands?

PTM · The supplied UniProt coordinates list N-acetylmethionine at position 1, phosphoserine at 11, and N6-lactoyllysine at 108. These sites do not, by themselves, demonstrate a detectable band shift. Coordinates refer to the supplied canonical sequence; isoform 2 lacks its first 15 residues, so its numbering differs.

Does this guide establish induction of TEAD1?

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 TEAD1?

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 M03263-1 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 nuclear TEAD1 be quantified?

Quantitation · UniProt places TEAD1 in the nucleus. For nuclear extracts, compare equal nuclear protein inputs and normalize the TEAD1 signal to a suitable nuclear loading reference. Keep the same band definition across samples, especially if more than one band appears.

Why might TEAD1 appear near 52 kDa instead of 47.9 kDa?

Interpretation · The supplied apparent band is about 52 kDa, while the predicted mass is 47.9 kDa. Confirm the band with a TEAD1-specific control; the listed features alone do not explain the difference or establish a visible shift.

Consider isoform 2 because it lacks two canonical sequence segments, residues 1–15 and 234–291. Its K110-to-KVTSM change also alters the sequence. A lower band is not proof of isoform identity; verify that the antibody recognizes the relevant sequence and use an isoform-specific control if available.

UniProt reports interactions with YAP1 and WWTR1/TAZ. Those interactions alone do not identify an additional TEAD1 band. Confirm unexpected bands with a TEAD1-specific control before assigning them to an isoform, modification, or interacting protein.

Boster reagents

TEAD1 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 TEAD1 using anti-TEAD1 antibody (M03263-1). Electrophoresis was performed on a 5-20% SDS-PAGE gel at 70V (Stacking gel) / 90V (Resolving gel) for 2-3 hours. The sample well of each lane was loaded with 30 ug of sample under reducing conditions. Lane 1: human Hela whole cell lysates, Lane 2: human SiHa 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-TEAD1 antigen affinity purified monoclonal antibody (M03263-1) at 1:1000 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 Enhanced Chemiluminescent detection (ECL) kit (Catalog # EK1002) with Tanon 5200 system. A specific band was detected for TEAD1 at approximately 52 kDa. The expected band size for TEAD1 is at 48 kDa.

Anti-TEAD1 Rabbit Monoclonal Antibody

Cat # M03263-1

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Real WB data Western blot analysis of TEAD1 expression in 293T cell lysate.

Anti-TEAD1 Rabbit Monoclonal Antibody

Cat # M03263-2

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Real WB data

Western blot analysis of TEF1/TEAD-1/TEAD1 using anti-TEF1/TEAD-1/TEAD1 antibody (A03263-2). Electrophoresis was performed on a 5-20% SDS-PAGE gel at 70V (Stacking gel) / 90V (Resolving gel) for 2-3 hours. The sample well of each lane was loaded with 30ug of sample under reducing conditions. Lane 1: human Hela whole cell lysates, Lane 2: human A431 whole cell lysates, Lane 3: human PC-3 whole cell lysates. After Electrophoresis, proteins were transferred to a Nitrocellulose membrane at 150mA 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-TEF1/TEAD-1/TEAD1 antigen affinity purified polyclonal antibody (Catalog # A03263-2) 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 Enhanced Chemiluminescent detection (ECL) kit (Catalog # EK1002) with Tanon 5200 system. A specific band was detected for TEF1/TEAD-1/TEAD1 at approximately 50KD. The expected band size for TEF1/TEAD-1/TEAD1 is at 50KD.

Anti-TEF1/TEAD-1/TEAD1 Antibody Picoband®

Cat # A03263-2

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Three the supplier anti-TEAD1 antibodies have supplied Western blot images. M03263-1 and A03263-2 provide detailed human cell lysate captions and reported band sizes; M03263-2 has a brief 293T caption. The images document tested samples, not validation across every listed species.

Which to pick: For human lysates, A03263-2 shows HeLa, A431, and PC-3 at approximately 50 kDa; M03263-1 shows HeLa and SiHa at approximately 52 kDa (48 kDa expected). M03263-2 shows 293T. For mouse or rat, only the two monoclonals list reactivity.

Source: BosterBio TEAD1 gene-info card — filtered to Western-blot-capable antibodies; each card shows that product's actual WB validation figure.

References

UniProt Consortium. UniProt entry P28347.
Human Protein Atlas. TEAD1 tissue expression.
PMC12190821 — target-verified WB comparison
PMC13232097 — target-verified WB comparison
PMC5735023 — target-verified WB comparison