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- Table of Contents
TET3 / Methylcytosine dioxygenase TET3 · Western blot design guide
Design a Western Blot for TET3
Real validated TET3 Western blot protocols, expected-band and isoform facts, troubleshooting for weak or shifted signal, and recommended anti-TET3 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
TET3 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 | ~193.7 kDa | |
| Gel | 8–10% (standard starting point) | |
| Positive control | Esophagus (IHC candidate; verify WB) +4 more | |
| Negative control | Adipose tissue (IHC candidate; verify WB) |
Important caveats
Reasons your observed band may differ from the expected size.
| PTM | Ubl conjugation | |
| Caveat | Blocking-peptide control | |
| Gene-set association | MSigDB C7 membership | |
| Isoform | 3 isoform(s) |
Section 1
Real Curated TET3 Western Blot Protocols
The A03371 protocol combines labelled catalog values with standard starting conditions. Published comparisons retain their own sample, reagent and detection scope.
| Sample / lysate | SK-N-SH cell lysate (catalog A03371) |
| Gel % | 8–10% (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 | Western blot analysis of TET3 in SK-N-SH cell lysate with TET3 antibody at 1 μg/ml in (A) the absence and (B) the presence of blocking peptide (catalog A03371) |
| Primary antibody | A03371 · 1 μg/ml (catalog A03371) |
| 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 TET3 Western Blot Band Size?
TET3 has a predicted mass of 193.7 kDa; listed splice isoforms may affect migration, but no empirical band size or distinct isoform bands are demonstrated.
| Single band near 193.7 kDa | consistent with predicted TET3 mass; confirm identity with controls |
| Several bands at different positions | could reflect isoforms 4, 2, and 3; their migration is unreported |
| Band below 193.7 kDa | could be a TET3 isoform, but its identity is unconfirmed |
| Band above 193.7 kDa | could be a TET3 isoform, but its identity is unconfirmed |
💡Expected TET3 appearanceUniProt predicts 193.7 kDa for TET3; no empirical band size or isoform migration is supplied, so identify any detected band with antibody blocking or another band-identity control.
| UniProt predicted mass | places the reference band near 193.7 kDa |
| Splice isoform 4 | may differ in apparent size; its direction and magnitude are unreported |
| Splice isoform 2 | may differ in apparent size; its direction and magnitude are unreported |
| Splice isoform 3 | may differ in apparent size; its direction and magnitude are unreported |
| Situation | Likely cause | Next action |
|---|---|---|
| No band in lysate | nuclear or chromosome-associated TET3 may be poorly recovered | check nuclear or chromatin-enriched material and a positive control |
| Band higher than expected | an isoform is possible, but no higher-migrating form is documented | confirm antibody specificity with blocking peptide and compare an independent antibody |
| Band lower than expected | an isoform is possible, but no lower-migrating form is documented | confirm antibody specificity with blocking peptide and compare an independent antibody |
| Multiple bands | splice isoforms 4, 2, and 3 are listed, but distinct bands are unverified | check which bands disappear with blocking peptide and compare an independent antibody |
| Weak or no signal | nuclear or chromosome-associated TET3 may be underrepresented in the sample | check sample recovery with nuclear or chromatin enrichment and a positive control |
Experiment evidence
Western blot images showing TET3 band
A03371 · TET3
← swipe / scroll for more images →
Real WB data behind the band-size expectations in this section.
Sample controls for TET3 Western blot
🧪HPA-IHC candidate guidance (verify in WB): For positive controls for TET3 in Western blot, you can use esophagus tissue, which HPA scores as high.
Positive control: Esophagus (IHC candidate; verify WB)
Negative control: Adipose tissue (IHC candidate; verify WB)
Loading controls: Run GAPDH, β-actin, and a total-protein stain such as Ponceau alongside the samples.
⚠️Feasibility: TET3 can localize to the nucleus, so use lysates that retain nuclear proteins.
HPA tissue expression evidence for TET3
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 |
|---|---|---|---|---|
| Esophagus | squamous epithelial cells | High | Protein (IHC) | HPA → |
| Oral mucosa | squamous epithelial cells | High | Protein (IHC) | HPA → |
| Placenta | trophoblastic cells | High | Protein (IHC) | HPA → |
| Skin | keratinocytes | High | Protein (IHC) | HPA → |
| Stomach | glandular cells | High | Protein (IHC) | HPA → |
Lower expression tissues · IHC evidence, not confirmed WB-negative controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| Adipose tissue | adipocytes | Not detected | Protein (IHC) | HPA → |
| Hippocampus | glial cells | Not detected | Protein (IHC) | HPA → |
| Liver | cholangiocytes | Not detected | Protein (IHC) | HPA → |
| Parathyroid gland | glandular cells | Not detected | Protein (IHC) | HPA → |
| Seminal vesicle | glandular cells | Not detected | Protein (IHC) | HPA → |
Section 3
Advanced TET3 Western Blot Tips
Deeper troubleshooting and optimisation questions for TET3, answered from its protein features.
Should the TET3 band appear at exactly 193.7 kDa?
Band shift · 193.7 kDa is the predicted mass of the 1,795-residue canonical protein. No observed band position is supplied, so use it as a reference rather than an exact apparent mass. The listed features do not establish a visible shift.
Could TET3 isoforms produce bands at different sizes?
Isoforms · Yes. In UniProt canonical-sequence coordinates, isoform 3 lacks residues 863–1795, and isoform 2 lacks residues 1575–1690. Isoform 4 is listed without a sequence change here. Antibody epitope location matters: an antibody against a missing region would not detect that isoform.
Do the supplied features identify a TET3 modification that shifts its band?
PTM · Ubl conjugation and isopeptide bond appear as keywords, and TET3 interacts with OGT. No modified-residue coordinates are supplied. These features do not establish which modification, if any, causes a visible shift; OGT interaction alone does not establish TET3 glycosylation.
Does this guide establish induction of TET3?
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 TET3 Western blot?
Transfer · TET3's canonical predicted mass is 193.7 kDa. Choose and check a transfer setup capable of retaining and transferring proteins near 194 kDa. Confirm transfer at that size before interpreting a weak or absent band; the supplied features do not specify a membrane, buffer, or transfer time.
How should blocking be optimized?
Blocking · Standard workflow guidance: follow the A03371 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 TET3 localization guide Western blot quantitation?
Quantitation · TET3 is annotated in the nucleus, cytoplasm, and chromosome, with stage-dependent localization during preimplantation development. Compare like fractions and stages, using a loading reference appropriate to each fraction. A change in one fraction alone need not represent a change in total TET3.
How should unexpected TET3 bands be interpreted?
Interpretation · Check whether a band could match the sequence deletions annotated for isoforms 2 or 3, and whether the antibody epitope remains in each isoform. The supplied features do not provide observed band positions, so they cannot establish the identity of an unexpected band. Use an independent antibody epitope or other identity check before assigning it.
Boster reagents
TET3 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-TET3 Antibody
Cat # A03371
the supplier A03371 is an anti-TET3 antibody listed for Western blotting with human, mouse, and rat reactivity. Its WB image shows SK-N-SH cell lysate at 1 μg/ml, with and without blocking peptide; the supplied evidence does not show other samples.
Which to pick: A03371 is the only listed TET3 antibody. It has a WB image using SK-N-SH lysate and is listed as reactive with human, mouse, and rat; the image documents that sample and condition only.
Source: BosterBio TET3 gene-info card — filtered to Western-blot-capable antibodies; each card shows that product's actual WB validation figure.
References
- UniProt Consortium. UniProt entry O43151.
- Human Protein Atlas. TET3 tissue expression.
- PMC13182592 — target-verified WB comparison
- PMC13169958 — target-verified WB comparison
- PMC10904493 — target-verified WB comparison