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TCF7L2 / Transcription factor 7-like 2 · Western blot design guide
Design a Western Blot for TCF7L2
Real validated TCF7L2 Western blot protocols, expected-band and isoform facts, troubleshooting for weak or shifted signal, and recommended anti-TCF7L2 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
TCF7L2 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 | ~67.9 kDa | |
| Gel | 12–15% (standard starting point) | |
| Positive control | Adrenal gland (IHC candidate; verify WB) +4 more | |
| Negative control | Suggested KO / knockdown lysate |
Important caveats
Reasons your observed band may differ from the expected size.
| PTM | Phosphorylated + Ubl conjugation | |
| Caveat | Modification-state controls | |
| Gene-set association | MSigDB Hallmark membership | |
| Isoform | 17 isoform(s) |
Section 1
Real Curated TCF7L2 Western Blot Protocols
The M00431 protocol combines labelled catalog values with standard starting conditions. Published comparisons retain their own sample, reagent and detection scope.
| Sample / lysate | Jurkat cell lysate (catalog M00431) |
| 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 | M00431; 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 TCF7L2 Western Blot Band Size?
TCF7L2 has a predicted mass of 67.9 kDa; isoforms could affect band position, but distinct migration has not been demonstrated here.
| Band near 67.9 kDa | Consistent with the predicted TCF7L2 mass; identity needs confirmation |
| Several bands at different positions | Could reflect TCF7L2 isoforms if band identity is confirmed |
| Weak band in whole-cell lysate | Nuclear localization may limit the detectable signal |
| Stronger band in a nuclear fraction | Consistent with TCF7L2 nuclear localization |
💡Expected TCF7L2 appearanceUniProt predicts 67.9 kDa, but no empirical band size is supplied; confirm any candidate band with ordinary antibody and target-identity controls.
| UniProt predicted mass | Places the reference expectation near 67.9 kDa without establishing apparent migration |
| Splice isoforms 1, 2, 3, 4, 5 and 6 | May differ in size; individual masses and migration are not supplied |
| Splice isoforms 7, 8, 9, 10, 11 and 12 | May differ in size; individual masses and migration are not supplied |
| Splice isoforms 13, 14, 15, 16 and 17 | May differ in size; individual masses and migration are not supplied |
| Situation | Likely cause | Next action |
|---|---|---|
| No band in lysate | TCF7L2 is nuclear and may be poorly recovered or diluted in whole-cell lysate | Check nuclear extraction and test a nuclear-enriched fraction |
| Multiple bands | TCF7L2 has 17 named splice isoforms, but their migration is unspecified | Confirm candidate bands with target depletion or an independent antibody |
| Band higher than expected | Isoform identity or another cause of altered migration is unverified | Check target identity and compare with the predicted 67.9 kDa reference |
| Band lower than expected | Isoform identity or fragment formation is unverified | Confirm target identity and inspect sample handling for proteolysis |
| Weak or no signal | Nuclear localization may reduce signal in whole-cell lysate | Enrich the nuclear fraction and verify extraction quality |
Experiment evidence
Western blot images showing TCF7L2 band
M00431 · TCF7L2
A00431-1 · TCF7L2
← swipe / scroll for more images →
Real WB data behind the band-size expectations in this section.
Sample controls for TCF7L2 Western blot
🧪HPA-IHC candidate guidance (verify in WB): For positive controls for TCF7L2 in Western blot, you can use adrenal gland lysate, which HPA rates High.
Positive control: Adrenal gland (IHC candidate; verify WB)
Negative control: Suggested KO / knockdown lysate
Loading controls: Run GAPDH, β-actin, and a total-protein stain alongside the samples.
⚠️Feasibility: No Not-detected tissue is listed, so use siRNA knockdown or a KO line as the negative control.
HPA tissue expression evidence for TCF7L2
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 | High | Protein (IHC) | HPA → |
| Appendix | glandular cells | High | Protein (IHC) | HPA → |
| Bone marrow | hematopoietic cells | High | Protein (IHC) | HPA → |
| Breast | glandular cells | High | Protein (IHC) | HPA → |
| Bronchus | respiratory epithelial cells | High | Protein (IHC) | HPA → |
Lower expression tissues · IHC evidence, not confirmed WB-negative controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| Adipose tissue | adipocytes | Medium | Protein (IHC) | HPA → |
| Heart muscle | cardiomyocytes | Medium | Protein (IHC) | HPA → |
| Soft tissue | fibroblasts | Medium | Protein (IHC) | HPA → |
| Spleen | cells in red pulp | Medium | Protein (IHC) | HPA → |
| Adrenal gland | glandular cells | High | Protein (IHC) | HPA → |
Section 3
Advanced TCF7L2 Western Blot Tips
Deeper troubleshooting and optimisation questions for TCF7L2, answered from its protein features.
How should TCF7L2 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.
How can TCF7L2 isoforms affect band interpretation?
Isoforms · UniProt lists 17 isoforms. For example, isoforms 9, 11, 14 and 15 lack canonical residues 466–619, while isoform 17 lacks residues 7–290. Different isoforms could therefore produce bands of different sizes. Check which isoforms the sample expresses before assigning bands by size.
An antibody against canonical residues 466–619 may miss isoforms 9, 11, 14 and 15, which lack that region, and other isoforms with C-terminal changes. Isoform 17 lacks canonical residues 7–290. Map the antibody epitope against the listed isoform sequences when comparing bands or samples.
Do not assume so. Isoform 17 lacks canonical residues 7–290, which include Thr201 and Thr212; isoform 11 replaces sequence beginning at canonical residue 184. Check the relevant isoform sequence before interpreting a phospho-specific signal. The listed coordinates use UniProt canonical numbering.
An antibody against canonical residues 466–619 may miss isoforms 9, 11, 14 and 15, which lack that region, and other isoforms with C-terminal changes. Isoform 17 lacks canonical residues 7–290. Map the antibody epitope against the listed isoform sequences when comparing bands or samples.
Do not assume so. Isoform 17 lacks canonical residues 7–290, which include Thr201 and Thr212; isoform 11 replaces sequence beginning at canonical residue 184. Check the relevant isoform sequence before interpreting a phospho-specific signal. The listed coordinates use UniProt canonical numbering.
Can NLK phosphorylation cause a TCF7L2 band shift?
PTM · UniProt lists phosphothreonine at canonical TCF7L2 residues 201 and 212, both attributed to NLK. These sites alone do not establish a visible shift. A phosphatase-treated comparison can help assess whether phosphorylation contributes to an observed mobility difference. Keep UniProt canonical numbering explicit when comparing antibody or paper site labels.
Does this guide establish induction of TCF7L2?
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 TCF7L2?
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 M00431 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 multiple TCF7L2 bands be quantified?
Quantitation · Specify which band or bands are measured and use the same selection across samples. UniProt lists 17 isoforms with several large sequence differences, so separate bands cannot be assigned to isoforms from size alone. Check antibody epitope coverage and report whether quantitation represents one candidate isoform or combined TCF7L2 signal.
Why might TCF7L2 bands differ from the predicted 67.9 kDa?
Interpretation · The 67.9 kDa prediction refers to the canonical protein. UniProt lists 17 isoforms with substantial sequence differences, including C-terminal deletions. No observed band position is supplied, and these features alone do not establish an apparent mass or visible shift. Compare each band with the sequence of the isoform your antibody can detect.
UniProt places TCF7L2 in the nucleus, including PML nuclear bodies, and also describes a diffuse nuclear pattern. A nuclear fraction is therefore relevant when assessing detection. Compare equivalent fractions across samples, since differing nuclear recovery could affect band intensity.
UniProt places TCF7L2 in the nucleus, including PML nuclear bodies, and also describes a diffuse nuclear pattern. A nuclear fraction is therefore relevant when assessing detection. Compare equivalent fractions across samples, since differing nuclear recovery could affect band intensity.
Boster reagents
TCF7L2 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-TCF7L2 Rabbit Monoclonal Antibody
Cat # M00431
Real WB data
Anti-Transcription factor 7-like 2 TCF7L2 Antibody
Cat # A00431-1
Both listed anti-TCF7L2 antibodies report Human, Mouse, and Rat reactivity and show Western blot results from Jurkat cell lysates. The supplied evidence does not document Western blot results in mouse or rat samples.
Which to pick: M00431 and A00431-1 both have Jurkat Western blot images and the same reported reactivity. Choose A00431-1 if a documented starting dilution helps: its image caption specifies 1:1,000. The supplied caption for M00431 does not specify a dilution.
Source: BosterBio TCF7L2 gene-info card — filtered to Western-blot-capable antibodies; each card shows that product's actual WB validation figure.
References
- UniProt Consortium. UniProt entry Q9NQB0.
- Human Protein Atlas. TCF7L2 tissue expression.
- PMC9208512 — target-verified WB comparison
- PMC11736883 — target-verified WB comparison
- PMC2926262 — target-verified WB comparison