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TET2 / Methylcytosine dioxygenase TET2 · Western blot design guide
Design a Western Blot for TET2
Real validated TET2 Western blot protocols, expected-band and isoform facts, troubleshooting for weak or shifted signal, and recommended anti-TET2 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
TET2 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 | ~223.8 kDa | |
| Gel | 4–12% gradient (standard starting point) | |
| Positive control | Adipose tissue (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 + Acetylated | |
| Caveat | Phosphorylation-state controls | |
| Gene-set association | MSigDB C7 membership | |
| Isoform | 3 isoform(s) |
Section 1
Real Curated TET2 Western Blot Protocols
The A00191-1 protocol combines labelled catalog values with standard starting conditions. Published comparisons retain their own sample, reagent and detection scope.
| Sample / lysate | 3T3 cell lysate (catalog A00191-1) |
| Gel % | 4–12% gradient (standard starting point) |
| Load | 20–30 µg total protein per lane; optimize for abundance (standard starting point) |
| Transfer | Wet/tank transfer; optimize duration (standard starting point) |
| Membrane | 0.45 µm PVDF (standard starting point) |
| Blocking | 5% milk or 5% BSA in TBST (standard starting point) |
| Primary antibody | A00191-1 · 1 μg/ml (catalog A00191-1) |
| 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 TET2 Western Blot Band Size?
TET2 is predicted at 223.8 kDa; splice isoforms could affect migration, but no empirical band size or distinct isoform pattern is supplied.
| Band near 223.8 kDa | Candidate full-length TET2 at its predicted mass; confirm identity with controls |
| Additional discrete bands | Could reflect isoforms 1, 2, and 3 if they migrate differently; their band positions are unknown |
| Weak band in cytoplasmic fraction | Consistent with TET2 localization to the nucleus and chromosomes |
| Band enriched in chromatin fraction | Consistent with TET2 chromosome localization |
💡Expected TET2 appearanceTET2 has a predicted mass of 223.8 kDa, but no empirical band size is supplied; confirm any candidate band with a positive control and TET2 depletion.
| Predicted TET2 mass | Full-length sequence predicts 223.8 kDa; apparent migration is unverified |
| Splice isoform 1 | Its relative band size is not supplied |
| Splice isoform 2 | It may differ from other isoforms, but its relative band size is not supplied |
| Splice isoform 3 | It may differ from other isoforms, but its relative band size is not supplied |
| Situation | Likely cause | Next action |
|---|---|---|
| No band in lysate | Nuclear or chromatin-associated TET2 may be poorly recovered | Check nuclear and chromatin fractions with a TET2-positive control |
| Band higher than expected | The supplied features do not establish a higher migrating TET2 species | Check band identity with TET2 depletion and a positive control |
| Band lower than expected | Alternative splicing is possible, but isoform masses are unknown | Check band identity with TET2 depletion and an isoform-aware reagent |
| Multiple bands | Isoforms 1, 2, and 3 may contribute, but distinct migration is unverified | Compare bands with TET2 depletion and an isoform-aware reagent |
| Weak or no signal | Nuclear and chromatin localization may limit recovery in the tested fraction | Assess nuclear extraction and compare with a TET2-positive sample |
Experiment evidence
Western blot images showing TET2 band
A00191-1 · TET2
A00191-2 · TET2
A00191-3 · TET2
A00191 · TET2
← swipe / scroll for more images →
Real WB data behind the band-size expectations in this section.
Sample controls for TET2 Western blot
🧪HPA-IHC candidate guidance (verify in WB): For positive controls for TET2 in Western blot, you can use adipose tissue lysate, which has high HPA expression.
Positive control: Adipose tissue (IHC candidate; verify WB)
Negative control: Suggested KO / knockdown lysate
Loading controls: Run GAPDH, β-actin, and a total-protein stain such as Ponceau alongside the samples.
⚠️Feasibility: HPA lists no tissue with undetected expression, so use TET2 knockdown or a KO line for a negative control.
HPA tissue expression evidence for TET2
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 |
|---|---|---|---|---|
| Adipose tissue | adipocytes | High | Protein (IHC) | HPA → |
| Adrenal gland | glandular cells | High | Protein (IHC) | HPA → |
| Bone marrow | hematopoietic cells | High | Protein (IHC) | HPA → |
| Breast | adipocytes | 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 |
|---|---|---|---|---|
| Appendix | glandular cells | Medium | Protein (IHC) | HPA → |
| Cerebral cortex | endothelial cells | Medium | Protein (IHC) | HPA → |
| Endometrium | cells in endometrial stroma | Medium | Protein (IHC) | HPA → |
| Epididymis | glandular cells | Medium | Protein (IHC) | HPA → |
| Liver | cholangiocytes | Medium | Protein (IHC) | HPA → |
Section 3
Advanced TET2 Western Blot Tips
Deeper troubleshooting and optimisation questions for TET2, answered from its protein features.
How should TET2 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 do TET2 isoforms affect antibody selection?
Isoforms · UniProt lists isoforms 1, 2, and 3. In UniProt coordinates, isoform 2 replaces residues 1137–1165 and lacks 1166–2002; isoform 3 replaces 1137–1194 and lacks 1195–2002. Check whether the antibody epitope lies in a shared region or in sequence absent from the isoform you want to measure.
Which listed TET2 modifications matter when interpreting bands?
PTM · UniProt lists phosphoserine at positions 15, 75, 99, 1107, and 1109, plus asymmetric dimethylarginine at 1682. These are UniProt coordinates; antibody or paper numbering may differ. Chromatin localization depends on monoubiquitination at Lys-1299. None of these features alone demonstrates a visible band shift.
Does this guide establish induction of TET2?
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.
What transfer method to use for TET2 Western blot?
Transfer · Canonical TET2 is predicted to be 223.8 kDa. Choose a transfer setup validated for proteins of this size, then check transfer near 224 kDa using a molecular weight marker or total-protein stain. The supplied features do not specify transfer conditions.
How should blocking be optimized?
Blocking · Standard workflow guidance: follow the A00191-1 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 TET2 bands be quantified across samples?
Quantitation · Measure the same band and isoform across samples. TET2 is found in the nucleus and on chromosomes, and its chromatin localization depends on monoubiquitination at Lys-1299. Keep the sample fraction consistent and normalize against an appropriate loading measure for that fraction.
Why might TET2 migrate differently from its predicted mass?
Interpretation · Canonical TET2 has a predicted mass of 223.8 kDa. Its listed modifications and alternative isoforms are reasons to investigate migration, but the features alone do not establish a visible shift or an apparent band size. Compare the band with a molecular weight marker and verify which isoform the antibody detects.
Consider the listed shorter isoforms: isoform 2 lacks canonical residues 1166–2002, and isoform 3 lacks 1195–2002, using UniProt coordinates. Check whether the antibody epitope is retained. The supplied features do not establish that any particular lower band is a TET2 isoform.
Consider the listed shorter isoforms: isoform 2 lacks canonical residues 1166–2002, and isoform 3 lacks 1195–2002, using UniProt coordinates. Check whether the antibody epitope is retained. The supplied features do not establish that any particular lower band is a TET2 isoform.
Boster reagents
TET2 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-TET2 Antibody
Cat # A00191-1
Real WB data
Anti-TET2 Antibody
Cat # A00191-2
Real WB data
Anti-TET2 Antibody
Cat # A00191-3
Real WB data
Anti-TET2 Antibody
Cat # A00191
Four the supplier anti-TET2 antibodies have Western blot images. The supplied captions show 3T3, THP-1, MCF-7, and SK-N-SH lysates, plus unspecified cell lines. These are example test conditions; no publication or independent validation evidence is supplied.
Which to pick: For human THP-1 or MCF-7 lysates, A00191-3 has a detailed blot caption; for SK-N-SH, consider A00191. For 3T3 lysate, consider A00191-1. A00191-2 lists Human, Mouse, and Rat reactivity, but its caption does not identify the cell lines.
Source: BosterBio TET2 gene-info card — filtered to Western-blot-capable antibodies; each card shows that product's actual WB validation figure.
References
- UniProt Consortium. UniProt entry Q6N021.
- Human Protein Atlas. TET2 tissue expression.
- PMC7186610 — target-verified WB comparison
- PMC6424975 — target-verified WB comparison
- PMC10910280 — target-verified WB comparison