This website uses cookies to ensure you get the best experience on our website.
- Table of Contents
TEF / Thyrotroph embryonic factor · Western blot design guide
Design a Western Blot for TEF
Real validated TEF Western blot protocols, expected-band and isoform facts, troubleshooting for weak or shifted signal, and recommended anti-TEF 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
TEF 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 | ~33.2 kDa | |
| Gel | 12–15% (standard starting point) | |
| Negative control | Suggested KO / knockdown lysate |
Important caveats
Reasons your observed band may differ from the expected size.
| PTM | Phosphorylated | |
| Caveat | Phosphorylation-state controls | |
| Gene-set association | MSigDB C7 membership | |
| Isoform | 2 isoform(s) |
Section 1
Real Curated TEF Western Blot Protocols
The A05436 protocol combines labelled catalog values with standard starting conditions. Published comparisons retain their own sample, reagent and detection scope.
| Sample / lysate | Target-positive lysate and matched negative control (standard starting point) |
| 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 | A05436; 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 TEF Western Blot Band Size?
TEF has a predicted monomer mass of 33.2 kDa; isoforms, phosphorylation, or retained dimers could affect appearance, but their migration is unverified.
| Band near 33.2 kDa | Consistent with the predicted TEF monomer; confirm identity with controls |
| Additional band at a different mass | Could reflect isoform 1 or 2; their migration is not established |
| Band near twice the monomer size | Could reflect a retained TEF homodimer if it survives sample preparation |
| No distinct phospho doublet | Phosphoserine 32 need not produce a visible mobility change |
💡Expected TEF appearanceUniProt predicts a 33.2 kDa TEF monomer, but no empirical band size is supplied; confirm any band near that position with ordinary band-identity controls.
| Predicted TEF monomer mass | Provides a 33.2 kDa reference, not a validated migration position |
| Isoform 1 | May differ in size from isoform 2; its mass is not supplied |
| Isoform 2 | May differ in size from isoform 1; its mass is not supplied |
| TEF homodimer | Could appear near twice the monomer size if retained during electrophoresis |
| Situation | Likely cause | Next action |
|---|---|---|
| No band in lysate | Nuclear TEF may be poorly recovered | Check a nuclear fraction and a positive control |
| Band higher than expected | A TEF homodimer may have persisted | Compare fully denatured samples with the original preparation |
| Band lower than expected | Isoforms 1 and 2 may differ in size | Check isoform recognition and confirm identity with TEF depletion |
| Multiple bands | TEF has two isoforms and phosphoserine 32, but distinct migration is unproven | Use TEF depletion and, for phospho-dependent bands, phosphatase treatment |
| Weak or no signal | Nuclear TEF may be underrepresented in the sample | Verify nuclear extraction and test a positive control |
Experiment evidence
Western blot images showing TEF band
A05436 · TEF
← swipe / scroll for more images →
Real WB data behind the band-size expectations in this section.
Sample controls for TEF Western blot
🧪For positive controls for TEF in Western blot, you can use no HPA-supported positive sample because none is provided.
Positive control: No high/medium HPA tissue identified
Negative control: Suggested KO / knockdown lysate
Loading controls: Run GAPDH, β-actin, and a total-protein stain alongside the samples.
⚠️Feasibility: HPA provides no expression data, so tissue controls cannot be selected; use a TEF knockdown or KO line for specificity.
HPA tissue expression evidence for TEF
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 |
|---|---|---|---|---|
| No high/medium HPA tissues identified in the supplied evidence. | ||||
Lower expression tissues · IHC evidence, not confirmed WB-negative controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| No lower-expression tissue rows available in the supplied evidence. | ||||
Section 3
Advanced TEF Western Blot Tips
Deeper troubleshooting and optimisation questions for TEF, answered from its protein features.
How should TEF 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.
Could TEF isoforms produce different bands?
Isoforms · UniProt lists isoforms 1 and 2. In isoform 2, the canonical residues 1–52 are replaced by a different 22-residue sequence. Compare bands with isoform-specific sequence expectations; the supplied features give no measured band positions.
Choose an epitope outside the canonical residues 1–52, which are replaced in isoform 2. An antibody against that N-terminal segment may miss isoform 2. Check the antibody's mapped epitope against both sequences.
Choose an epitope outside the canonical residues 1–52, which are replaced in isoform 2. An antibody against that N-terminal segment may miss isoform 2. Check the antibody's mapped epitope against both sequences.
How should phosphorylation affect TEF band interpretation?
PTM · UniProt annotates phosphoserine at position 32 in the canonical sequence. That residue lies within the segment replaced in isoform 2. Phosphorylation is a possible source of band variation, but this annotation alone does not establish a visible shift.
Does this guide establish induction of TEF?
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 TEF?
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 A05436 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 TEF bands be quantified?
Quantitation · TEF is annotated as nuclear, so use a consistent nuclear sample preparation when comparing conditions. Decide whether the assay measures total TEF or a specific isoform, and quantify the corresponding bands consistently. The supplied features do not establish how TEF abundance changes between conditions.
Should TEF run at its predicted 33.2 kDa?
Interpretation · Use 33.2 kDa as the predicted reference for canonical TEF. No observed band size is supplied, so the features cannot establish where its band runs or explain a difference from the prediction.
Check isoform identity first: isoform 2 has a different N-terminal sequence. Also consider the annotated canonical phosphoserine at UniProt position 32, while recognizing that its presence does not prove a visible shift. TEF can dimerize with itself or DBP, but the supplied features do not establish that an unexpected blot band is a dimer.
Check isoform identity first: isoform 2 has a different N-terminal sequence. Also consider the annotated canonical phosphoserine at UniProt position 32, while recognizing that its presence does not prove a visible shift. TEF can dimerize with itself or DBP, but the supplied features do not establish that an unexpected blot band is a dimer.
Boster reagents
TEF 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-Thyrotroph embryonic factor TEF Antibody
Cat # A05436
The catalog reports anti-TEF antibody A05436 with reported Human, Mouse, and Rat reactivity. Its catalog includes a Western blot image described as analysis of cells, but the supplied caption does not identify the cell type or establish WB performance in each listed species.
Which to pick: A05436 is the only listed TEF antibody and has a WB image. Check whether its reported Human, Mouse, or Rat reactivity fits your sample; the supplied WB description does not specify the cells tested.
Source: BosterBio TEF gene-info card — filtered to Western-blot-capable antibodies; each card shows that product's actual WB validation figure.
References
- UniProt Consortium. UniProt entry Q10587.
- Human Protein Atlas. TEF tissue expression.
- PMC11308427 — target-verified WB comparison
- PMC3037971 — target-verified WB comparison