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TEAD2 · Western blot design guide
Design a Western Blot for TEAD2
Real validated TEAD2 Western blot protocols, expected-band and isoform facts, troubleshooting for weak or shifted signal, and recommended anti-TEAD2 WB antibodies. Everything you need to plan the experiment before you commit precious samples.
Last reviewed: May 2026 · Scientific review: Boster Bio technical team
TEAD2 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 | ~49.2 kDa | |
| Observed band | ~49 kDa | |
| Gel | 10–12% | |
| Negative control | siRNA / KO lysate |
Important caveats
Reasons your observed band may differ from the expected size.
| PTM | — | |
| Caveat | Alternative splicing isoforms | |
| Regulation | IL-2-induced | |
| Isoform | 4 isoform(s) |
Section 1
Real Curated TEAD2 Western Blot Protocols
Literature-validated Western blot parameters for TEAD2 — gel percentage, transfer, blocking, antibody incubation and detection, extracted from published methods.
| Sample / lysate | human Hela . 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-ETF/TEAD2 antigen affinity purified polyclonal antibody (Catalog # A09168-2) 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 ETF/TEAD2 at approximately 49KD. The expected band size for ETF/TEAD2 is at 49KD |
| Gel % | 10–12% |
| Load | 50ug |
| Transfer | nitrocellulose membrane, 150 mA, 50–90 min |
| Membrane | nitrocellulose |
| Blocking | 5% non-fat milk / TBS, 1.5 h RT |
| Primary antibody | 0.5 µg/mL |
| Primary incubation | overnight at 4 °C |
| Secondary antibody | goat anti-rabbit IgG-HRP, 1:5000 |
| Wash | TBS-0.1% Tween, 3 × 5 min |
| Detection | ECL |
| Exposure / imaging | Tanon 5200 |
| Observed band | 49 kDa |
Section 2
What Is the Expected TEAD2 Western Blot Band Size?
TEAD2 has a 49.2 kDa predicted mass and runs at ~49 kDa on Western blots, closely matching prediction since it lacks glycosylation, disulfide bonds, and proteolytic processing.
| Single band at ~49 kDa | Matches the unmodified 49.2 kDa predicted mass of TEAD2, consistent with no glycosylation or proteolytic processing |
| Band unchanged after glycosidase (e.g. PNGase F) treatment | TEAD2 has no annotated glycosylation sites, so its apparent mass is not glyco-dependent |
| No shift in band position between reducing and non-reducing gels | TEAD2 has no annotated disulfide bonds and does not form a covalent homodimer, so it runs as a monomer either way |
| No smaller cleaved fragment beneath the main band | TEAD2 has no signal peptide or propeptide, so there is no precursor-to-mature cleavage step to shrink the band |
| Closely spaced doublet or extra faint band near 49 kDa | TEAD2 has four annotated splice isoforms that can migrate as slightly different species around the same size |
| Band present in nuclear fraction but weak or absent in cytoplasmic fraction | TEAD2 is annotated as a nuclear protein, so it should be enriched in nuclear extracts rather than whole cytoplasm |
💡Expected TEAD2 appearanceTEAD2 typically appears as a single band at approximately 49 kDa, matching its 49.2 kDa predicted mass, since it has no glycosylation, disulfide bonds, or signal-peptide/propeptide cleavage; closely spaced isoform bands near this size are possible.
| Predicted molecular weight from UniProt (49.2 kDa) | Sets the baseline expectation, and matches the ~49 kDa band actually observed on Western blot |
| Absence of annotated glycosylation sites | Keeps the band close to its unmodified predicted mass rather than shifting or smearing higher |
| Absence of annotated disulfide bonds or homodimer formation | Band stays monomeric and does not double in apparent size under non-reducing conditions |
| Named splice isoforms 1, 2, 3, and 4 | Can produce additional closely spaced bands of slightly different apparent size around the main band |
| Absence of signal peptide or propeptide | No precursor-to-mature cleavage occurs, so no smaller processed fragment appears below the main band |
| Situation | Likely cause | Next action |
|---|---|---|
| No band in lysate | TEAD2 is a nuclear transcription factor, and whole-cell lysis that poorly recovers nuclear protein can under-represent it | Use a nuclear or whole-cell extraction method optimized for nuclear proteins and load sufficient total protein from a TEAD2-expressing sample |
| Multiple bands | Four annotated splice isoforms of TEAD2 can generate closely spaced species near the main band | Check whether the antibody epitope overlaps all isoforms and compare band pattern to the canonical isoform 1 size |
| Band higher than expected | Detection of a larger splice isoform rather than the canonical form | Compare the apparent size to the ~49 kDa canonical band and confirm the immunogen region is present in the detected isoform |
| Band lower than expected | Detection of a shorter splice isoform or partial degradation during nuclear lysate preparation | Add protease inhibitors during lysis and compare against the canonical ~49 kDa isoform 1 band |
| Weak or no signal | TEAD2 is a nuclear protein and inefficient nuclear lysis or low loading can under-represent it | Use a dedicated nuclear extraction buffer, increase protein loading, and extend primary antibody incubation |
| Fragments below expected size | Nuclear TEAD2 can undergo proteolytic degradation during sample preparation | Prepare lysates on ice with protease inhibitors and minimize freeze-thaw cycles before loading |
Experiment evidence
Western blot images showing TEAD2 band
A09168-2 · TEAD2
← swipe / scroll for more images →
Real WB data behind the band-size expectations in this section.
Sample controls for TEAD2 Western blot
🧪For positive controls for TEAD2 in Western blot, you can use lysate from a TEAD2-overexpressing cell line (e.g., HEK293 cells transiently transfected with a TEAD2 expression construct), since no HPA tissue or cell-line expression data are available for this protein.
Positive control: TEAD2-transfected HEK293 cells
Negative control: no confirmed negative tissue on record; use TEAD2 siRNA knockdown or CRISPR KO cells
Loading controls: Also probe for GAPDH, β-actin, and a total-protein stain (Ponceau, REVERT, or stain-free gel) as loading controls.
⚠️Feasibility: With no HPA expression data for TEAD2, a validated positive or negative tissue cannot be confirmed, so confirm antibody specificity using an overexpression system alongside siRNA knockdown or a knockout line.
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.
Positive expression · recommended positive controls
| Tissue | Cell type | Level | Evidence | Source |
|---|
Undetected expression · recommended negative controls
| Tissue | Cell type | Level | Evidence | Source |
|---|
Section 3
Advanced TEAD2 Western Blot Tips
Deeper troubleshooting and optimisation questions for TEAD2, answered from its protein features.
Does the observed TEAD2 band match its predicted size?
TEAD2's predicted mass is 49.2 kDa and the empirically observed band runs at approximately 49 kDa, showing no shift. This close agreement indicates minimal post-translational modification affecting migration, consistent with UniProt reporting no glycosylation, disulfide bonds, or modified residues for TEAD2. A band at ~49 kDa is expected on standard SDS-PAGE.
Why might multiple bands appear near 49 kDa?
UniProt lists four TEAD2 isoforms (1-4) generated by alternative splicing. These can produce closely spaced bands around the predicted 49.2 kDa mass. Confirm which isoform your antibody targets, since epitope location determines whether all splice variants are detected or only a subset.
Is TEAD2 expression regulated by Hippo signaling?
TEAD2 itself is a stable DNA-binding transcription factor, but its transcriptional activity depends on nuclear interaction with coactivators YAP1 and WWTR1/TAZ, effectors of the Hippo pathway. Total TEAD2 protein levels may stay constant while activity varies, so consider co-detecting YAP1/TAZ if assessing pathway activation rather than TEAD2 levels alone.
What blocking approach works best for TEAD2 detection?
Since TEAD2 is a nuclear DNA-binding protein without glycosylation or other extracellular modifications, standard 5% non-fat milk or BSA in TBST blocking is sufficient. Use BSA rather than milk if also probing phospho-specific YAP1/TAZ antibodies on the same membrane, as milk contains phosphoproteins that can raise background.
What transfer method to use for TEAD2 Western blot?
At 49.2 kDa predicted mass, TEAD2 transfers efficiently using standard wet or semi-dry transfer methods for mid-size proteins. Nuclear proteins can be prone to incomplete extraction, so ensure lysis buffer effectively solubilizes nuclear fractions before transfer; overly long transfer times are not required for this mass range.
Which loading control suits TEAD2 quantitation?
Because TEAD2 localizes to the nucleus, use a nuclear loading control such as Histone H3 or Lamin B1 rather than a whole-cell control like GAPDH, especially if performing nuclear/cytoplasmic fractionation to assess YAP1/TAZ-dependent TEAD2 activity.
What explains unexpected bands beyond the main TEAD2 signal?
Extra bands may reflect the four annotated TEAD2 splice isoforms rather than nonspecific binding. Bands could also represent TEAD2 in complex with YAP1 or WWTR1/TAZ if samples aren't fully denatured/reduced before loading. Verify complete sample denaturation and compare against isoform-specific epitope mapping before concluding nonspecific signal.
Boster reagents
Best TEAD2 Western Blot Antibodies
BosterBio's TEAD2 antibodies are among the best-performing WB antibodies on the market — well cited, thoroughly validated, and orthogonally cross-validated against negative tissues and complementary methods.
Real WB data
Anti-ETF/TEAD2 Antibody Picoband®
Cat # A09168-2
One Boster antibody is available for TEAD2 western blotting, A09168-2, shown here with its own WB validation image. Data beyond this single image (citations, independent replication, negative-control comparisons) are not provided, so claims are limited to what the image shows.
Which to pick: Only one option is listed for TEAD2: A09168-2, which includes a real WB image (SDS-PAGE, 5-20% gel). Choose it by default since it is the sole catalogued anti-TEAD2 WB antibody here; confirm species/lysate match before use.
Source: BosterBio TEAD2 gene-info card — filtered to Western-blot-capable antibodies; each card shows that product's actual WB validation figure.
References
- UniProt Consortium. UniProt entry Q15562.
- Human Protein Atlas. TEAD2 tissue expression.