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- Table of Contents
TCF12 / Transcription factor 12 · Western blot design guide
Design a Western Blot for TCF12
Real validated TCF12 Western blot protocols, expected-band and isoform facts, troubleshooting for weak or shifted signal, and recommended anti-TCF12 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
TCF12 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 | ~73 kDa | |
| Gel | 12–15% (standard starting point) | |
| Positive control | Adrenal gland (IHC candidate; verify WB) +4 more | |
| Negative control | Parathyroid gland (IHC candidate; verify WB) |
Important caveats
Reasons your observed band may differ from the expected size.
| PTM | Phosphorylated + Ubl conjugation | |
| Caveat | Phosphorylation-state controls | |
| Gene-set association | MSigDB C7 membership | |
| Isoform | 4 isoform(s) |
Section 1
Real Curated TCF12 Western Blot Protocols
The A02592 protocol combines labelled catalog values with standard starting conditions. Published comparisons retain their own sample, reagent and detection scope.
| Sample / lysate | HeLa cell lysate (catalog A02592) |
| 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 | A02592 · (A) 0.5 and (B) 1 μg/mL (catalog A02592) |
| 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 TCF12 Western Blot Band Size?
TCF12 is predicted at 73 kDa; isoforms and phosphorylation could affect migration, but no empirical band position or feature-driven shift is established.
| Band near 73 kDa | consistent with the predicted TCF12 size; confirm identity with controls |
| Several bands near the expected region | could reflect isoforms 1, 2, 3 and 4; their migration is not established |
| Closely spaced bands | could reflect phosphorylation; a visible shift is not established |
| Stronger band in nuclear extract than whole-cell lysate | consistent with TCF12's nuclear location |
💡Expected TCF12 appearanceTCF12 has a predicted mass of 73 kDa, but no empirical band size is supplied; confirm any candidate band with antibody specificity controls and nuclear enrichment.
| Predicted TCF12 mass | provides a 73 kDa reference, not a measured migration position |
| Isoforms 1, 2, 3 and 4 | may differ in size; their masses and band separation are not supplied |
| Phosphoserine at Ser47 | may affect migration; no shift is established |
| Phosphothreonine at Thr313 | may affect migration; no shift is established |
| Situation | Likely cause | Next action |
|---|---|---|
| No band in lysate | TCF12 is nuclear and may be poorly represented in the sampled lysate | check nuclear extraction and compare a nuclear fraction |
| Band higher than expected | phosphorylation is documented, but its effect on migration is unknown | compare phosphatase-treated samples and verify band identity |
| Band lower than expected | an isoform or degradation is possible; isoform masses are unknown | check sample integrity and antibody specificity |
| Multiple bands | four isoforms and phosphorylation are documented, but band separation is unknown | compare antibody reactivity and phosphatase-treated samples |
| Weak or no signal | nuclear TCF12 may be underrepresented in whole-cell lysate | enrich nuclei and check loading and antibody performance |
| Fragments below expected size | sample degradation is possible | prepare fresh lysate with protease inhibitors and verify band identity |
Experiment evidence
Western blot images showing TCF12 band
A02592 · TCF12
A02592-3 · TCF12
← swipe / scroll for more images →
Real WB data behind the band-size expectations in this section.
Sample controls for TCF12 Western blot
🧪HPA-IHC candidate guidance (verify in WB): For positive controls for TCF12 in Western blot, you can use adrenal gland tissue, which HPA scores as High.
Positive control: Adrenal gland (IHC candidate; verify WB)
Negative control: Parathyroid gland (IHC candidate; verify WB)
Loading controls: Run GAPDH, β-actin, and a total-protein stain such as Ponceau alongside the samples.
⚠️Feasibility: HPA reports a not-detected tissue, making a tissue negative control feasible for this nuclear protein.
HPA tissue expression evidence for TCF12
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 → |
| Bone marrow | hematopoietic cells | High | Protein (IHC) | HPA → |
| Breast | glandular cells | High | Protein (IHC) | HPA → |
| Bronchus | respiratory epithelial cells | High | Protein (IHC) | HPA → |
| Caudate | glial cells | High | Protein (IHC) | HPA → |
Lower expression tissues · IHC evidence, not confirmed WB-negative controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| Parathyroid gland | glandular cells | Not detected | Protein (IHC) | HPA → |
| Liver | hepatocytes | Low | Protein (IHC) | HPA → |
| Smooth muscle | smooth muscle cells | Low | Protein (IHC) | HPA → |
| Spleen | cells in red pulp | Low | Protein (IHC) | HPA → |
| Adipose tissue | adipocytes | Medium | Protein (IHC) | HPA → |
Section 3
Advanced TCF12 Western Blot Tips
Deeper troubleshooting and optimisation questions for TCF12, answered from its protein features.
Where should the main TCF12 band appear?
Band shift · The predicted mass is 73 kDa. No empirical band position is supplied, so use 73 kDa as a starting reference, not an expected apparent position. The listed isoforms and modifications alone do not establish a visible shift.
How could isoform 2 affect the TCF12 band?
Isoforms · Isoform 2 lacks UniProt canonical residues 1–170 and replaces residues 171–193 with a different sequence. It could differ in size and antibody recognition. Check whether the antibody epitope lies in these regions before assigning a band to isoform 2.
Isoform 4 replaces canonical residues 1–39 and lacks residues 40–275; isoform 3 changes the sequence at residue 396. Compare an antibody’s epitope with these regions before interpreting additional bands. The supplied features do not give observed positions for any isoform.
Isoform 4 replaces canonical residues 1–39 and lacks residues 40–275; isoform 3 changes the sequence at residue 396. Compare an antibody’s epitope with these regions before interpreting additional bands. The supplied features do not give observed positions for any isoform.
Which phosphorylation sites matter when interpreting TCF12 bands?
PTM · UniProt lists phosphoserine at 47, 67, 79, 98, 116, 333, 540, 558 and 559, and phosphothreonine at 313 and 557. These are canonical UniProt coordinates; paper or antibody numbering may differ. Their presence does not establish a visible mobility shift.
Does this guide establish induction of TCF12?
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 TCF12?
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 A02592 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 TCF12 signals be quantified across samples?
Quantitation · TCF12 is nuclear and has four listed isoforms. Use comparable nuclear samples and define consistently which band or bands enter the measurement. If multiple bands appear, report them separately unless their identities are established; the features alone do not identify an observed band.
How should unexpected TCF12 bands be interpreted?
Interpretation · First compare their positions with the 73 kDa predicted mass, then check whether the antibody epitope is retained in each isoform. Phosphorylation is also listed, but it does not by itself explain a band shift. Without an empirical band position or band identity, avoid assigning an unexpected band to a specific isoform or modification.
Boster reagents
TCF12 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-Transcription factor 12 TCF12 Antibody
Cat # A02592
Real WB data
Anti-HEB/TCF12 Antibody Picoband®
Cat # A02592-3
Both listed anti-TCF12 antibodies, A02592 and A02592-3, claim human, mouse, and rat reactivity and have Western blot images. A02592 is shown with HeLa lysate; A02592-3 shows human cell, rat, and mouse samples. For A02592-3, the reported band is approximately 85 kDa versus 73 kDa expected.
Which to pick: Choose A02592-3 for samples resembling its documented human cell lines or rat and mouse specimens. For HeLa lysate, A02592 also has an image at 0.5 and 1 μg/mL. The images show tested contexts, not performance across all samples.
Source: BosterBio TCF12 gene-info card — filtered to Western-blot-capable antibodies; each card shows that product's actual WB validation figure.
References
- UniProt Consortium. UniProt entry Q99081.
- Human Protein Atlas. TCF12 tissue expression.
- PMC7789671 — target-verified WB comparison
- PMC10792168 — target-verified WB comparison
- PMC6735379 — target-verified WB comparison