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- Table of Contents
TFG / Protein TFG · Western blot design guide
Design a Western Blot for TFG
Real validated TFG Western blot protocols, expected-band and isoform facts, troubleshooting for weak or shifted signal, and recommended anti-TFG 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
TFG 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 | ~43.4 kDa | |
| Observed band | ~60 kDa | |
| Gel | 5–20% (catalog A02870-3) | |
| Positive control | Appendix (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 | Phosphorylated + Acetylated | |
| Caveat | Band identity controls | |
| Gene-set association | MSigDB C7 membership | |
| Isoform | 4 isoform(s) |
Section 1
Real Curated TFG Western Blot Protocols
The A02870-3 protocol combines labelled catalog values with standard starting conditions. Published comparisons retain their own sample, reagent and detection scope.
| Sample / lysate | human MCF-7, human Hela, human A549 (catalog A02870-3) |
| Gel % | 5–20% (catalog A02870-3) |
| Load | 30 ug; reducing conditions (catalog A02870-3) |
| Transfer | a nitrocellulose membrane at 150 mA for 50-90 minutes (catalog A02870-3) |
| Membrane | nitrocellulose membrane (catalog A02870-3) |
| Blocking | 5% non-fat milk/TBS for 1.5 hour at RT (catalog A02870-3) |
| Primary antibody | A02870-3 · 0.25 μg/mL (catalog A02870-3) |
| Primary incubation | overnight at 4°C (catalog A02870-3) |
| Secondary antibody | goat anti-rabbit IgG-HRP, 1:5000 (catalog A02870-3) |
| Secondary incubation | 1.5 hour at RT (catalog A02870-3) |
| Wash | TBS-0.1%Tween 3 times with 5 minutes each (catalog A02870-3) |
| Detection | ECL (catalog A02870-3) |
Section 2
What Is the Expected TFG Western Blot Band Size?
TFG is predicted at 43.4 kDa, while antibody QC reports ~60 kDa; the cause of the difference is not established.
| Band at ~60 kDa | TFG band reported in antibody QC; its identity still warrants controls |
| Band near 43.4 kDa | Near the predicted mass of TFG |
| Several bands at different positions | TFG isoforms 1, 2, 3, and 4 are possible contributors, but distinct migration is unestablished |
| Weak band in an ER-depleted fraction | Consistent with TFG localization at ER exit sites |
💡Expected TFG appearanceTFG has a predicted mass of 43.4 kDa and an empirical band at ~60 kDa; the cause of the difference is unestablished, so confirm band identity with appropriate controls.
| Predicted TFG mass | 43.4 kDa from the supplied UniProt sequence; apparent migration may differ |
| Isoform 1 | May differ in size from other named isoforms; its mass is unspecified |
| Isoform 2 | May differ in size from other named isoforms; its mass is unspecified |
| Isoform 3 | May differ in size from other named isoforms; its mass is unspecified |
| Isoform 4 | May differ in size from other named isoforms; its mass is unspecified |
| Situation | Likely cause | Next action |
|---|---|---|
| Band higher than expected | The empirical ~60 kDa band exceeds the 43.4 kDa prediction; the cause is unestablished | Compare with the reported ~60 kDa band and confirm identity using an independent antibody or TFG depletion |
| Band lower than expected | A different TFG isoform is possible, but isoform migration is unspecified | Check antibody epitope coverage and confirm identity by TFG depletion |
| Multiple bands | TFG has four named isoforms, but distinct bands are not established | Compare isoform expression and test which bands respond to TFG depletion |
| Weak or no signal | TFG localizes to ER exit sites; sample fraction and abundance may affect detection | Check ER-containing sample material and include a positive-control lysate |
| Fragments below expected size | Fragment identities are not established by the supplied features | Review sample handling and verify TFG specificity with an independent antibody or TFG depletion |
Experiment evidence
Western blot images showing TFG band
A02870-3 · TFG
M02870 · TFG
← swipe / scroll for more images →
Real WB data behind the band-size expectations in this section.
Sample controls for TFG Western blot
🧪HPA-IHC candidate guidance (verify in WB): For positive controls for TFG in Western blot, you can use appendix tissue lysate, which HPA rates as high for TFG.
Positive control: Appendix (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: HPA reports no detectable TFG in adipose tissue, providing a potential negative tissue control.
HPA tissue expression evidence for TFG
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 |
|---|---|---|---|---|
| Appendix | glandular cells | High | Protein (IHC) | HPA → |
| Cerebral cortex | glial cells | High | Protein (IHC) | HPA → |
| Colon | glandular cells | High | Protein (IHC) | HPA → |
| Duodenum | glandular cells | High | Protein (IHC) | HPA → |
| Gallbladder | 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 → |
| Cervix | glandular cells | Not detected | Protein (IHC) | HPA → |
| Esophagus | squamous epithelial cells | Not detected | Protein (IHC) | HPA → |
| Liver | cholangiocytes | Not detected | Protein (IHC) | HPA → |
| Lung | alveolar cells | Not detected | Protein (IHC) | HPA → |
Section 3
Advanced TFG Western Blot Tips
Deeper troubleshooting and optimisation questions for TFG, answered from its protein features.
How should TFG 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.
Which TFG isoforms could produce different bands?
Isoforms · UniProt lists four isoforms. Isoforms 2 and 4 lack residues 237–240; isoforms 3 and 4 replace residues 274–284 and lack residues 285–400. These sequence differences could affect band position and antibody recognition. The coordinates refer to the supplied canonical UniProt sequence.
Choose a validated epitope within canonical residues 1–236, which precede all listed alternative sequence changes. An epitope within residues 285–400 would miss isoforms 3 and 4. Check the antibody’s stated epitope against the canonical UniProt coordinates before interpreting missing bands.
Use an antibody recognizing a region shared by all four isoforms, such as canonical residues 1–236, and quantify resolved bands separately. Isoforms 3 and 4 lack residues 285–400, including the listed methylarginines at 385 and 400. Report which bands and antibody epitope were measured.
Choose a validated epitope within canonical residues 1–236, which precede all listed alternative sequence changes. An epitope within residues 285–400 would miss isoforms 3 and 4. Check the antibody’s stated epitope against the canonical UniProt coordinates before interpreting missing bands.
Use an antibody recognizing a region shared by all four isoforms, such as canonical residues 1–236, and quantify resolved bands separately. Isoforms 3 and 4 lack residues 285–400, including the listed methylarginines at 385 and 400. Report which bands and antibody epitope were measured.
Which TFG modifications matter when interpreting bands?
PTM · UniProt lists N-acetylmethionine at position 1, phosphoserine at 50 and 197, and omega-N-methylarginine at 385 and 400. These are canonical UniProt coordinates; antibody or paper numbering may differ. A modification site alone does not establish a visible mobility shift.
Does this guide establish induction of TFG?
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 TFG?
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 A02870-3 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 TFG be quantified?
Quantitation · Standard workflow guidance: quantify only a validated target band within the linear exposure range. Use consistent sample preparation and loading, retain biological replicates, and avoid interpreting saturation or loading differences as regulation.
Why might TFG appear near 60 kDa instead of 43.4 kDa?
Interpretation · The supplied apparent band is about 60 kDa, while the canonical sequence predicts 43.4 kDa. UniProt lists alternative isoforms and modifications, but these features alone do not explain the difference or establish a visible shift. Confirm the band with an independent TFG antibody or a TFG depletion control.
Lower bands could be assessed against the listed isoforms, particularly isoforms 3 and 4, which lack residues 285–400. TFG also self-associates into oligomeric complexes, so investigate higher bands with an independent antibody or TFG depletion control. Neither feature alone identifies an unexpected band.
Lower bands could be assessed against the listed isoforms, particularly isoforms 3 and 4, which lack residues 285–400. TFG also self-associates into oligomeric complexes, so investigate higher bands with an independent antibody or TFG depletion control. Neither feature alone identifies an unexpected band.
Boster reagents
TFG 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-TRK fused gene/TFG Antibody Picoband®
Cat # A02870-3
Real WB data
Anti-TRK fused gene TFG Rabbit Monoclonal Antibody
Cat # M02870
Two the supplier anti-TFG antibodies are listed for Western blotting, both with WB images and stated human, mouse, and rat reactivity. A02870-3 has a detailed caption covering samples from all three species, although its observed band is approximately 60 kDa versus an expected 43 kDa.
Which to pick: Choose A02870-3 if you want documented WB conditions and sample examples across human, rat, and mouse. M02870 is a rabbit monoclonal with a WB image captioned for HeLa lysate; its supplied caption gives no comparable conditions or other tested specimens.
Source: BosterBio TFG gene-info card — filtered to Western-blot-capable antibodies; each card shows that product's actual WB validation figure.
References
- UniProt Consortium. UniProt entry Q92734.
- Human Protein Atlas. TFG tissue expression.
- PMC3317498 — target-verified WB comparison
- PMC5310050 — target-verified WB comparison
- PMC12009431 — target-verified WB comparison