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- Table of Contents
TGFA / Protransforming growth factor alpha · Western blot design guide
Design a Western Blot for TGFA
Real validated TGFA Western blot protocols, expected-band and isoform facts, troubleshooting for weak or shifted signal, and recommended anti-TGFA 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
TGFA 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 | ~17 kDa | |
| Observed band | ~17 kDa | |
| Gel | 5–20% (catalog M01779-1) | |
| Positive control | Adrenal gland (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 | Glycosylated + Cleaved | |
| Caveat | Processing-state controls | |
| Gene-set association | MSigDB Hallmark membership | |
| Isoform | 5 isoform(s) |
Section 1
Real Curated TGFA Western Blot Protocols
The M01779-1 protocol combines labelled catalog values with standard starting conditions. Published comparisons retain their own sample, reagent and detection scope.
| Sample / lysate | human HepG2, human A431 (catalog M01779-1) |
| Gel % | 5–20% (catalog M01779-1) |
| Load | 30 ug; reducing conditions (catalog M01779-1) |
| Transfer | a nitrocellulose membrane at 150 mA for 50-90 minutes (catalog M01779-1) |
| Membrane | nitrocellulose membrane (catalog M01779-1) |
| Blocking | 5% non-fat milk/TBS for 1.5 hour at RT (catalog M01779-1) |
| Primary antibody | M01779-1 · 1:500 (catalog M01779-1) |
| Primary incubation | overnight at 4°C (catalog M01779-1) |
| Secondary antibody | goat anti-rabbit IgG-HRP, 1:500 (catalog M01779-1) |
| Secondary incubation | 1.5 hour at RT (catalog M01779-1) |
| Wash | TBS-0.1%Tween 3 times with 5 minutes each (catalog M01779-1) |
| Detection | ECL (catalog M01779-1) |
Section 2
What Is the Expected TGFA Western Blot Band Size?
TGFA is predicted at 17 kDa and observed at ~17 kDa; its listed processing, glycosylation, and isoforms do not establish other band positions.
| Band at ~17 kDa | Matches the reported TGFA band and predicted precursor mass; confirm identity with controls |
| Band below ~17 kDa | May represent TGFA processed from its signal peptide or propeptide; its migration is not established |
| Little or no band in whole-cell lysate | Secreted TGFA may be present in extracellular material |
| Several bands at different positions | Could reflect TGFA isoforms 1–5 or processing; distinct isoform bands are not established |
💡Expected TGFA appearanceTGFA has a predicted precursor mass of 17 kDa and a reported band at ~17 kDa in reducing whole-cell lysates; confirm band identity with an appropriate positive control or antibody validation.
| Predicted precursor mass | 17 kDa; the reported band is at ~17 kDa |
| N-linked glycosylation at Asn25 | May alter apparent migration, but a visible shift is not established |
| Signal peptide at residues 1–23 | Cleavage can make processed TGFA smaller than the precursor; no band size is supplied |
| Propeptide at residues 24–39 | Cleavage can further reduce size; no processed band size is supplied |
| Isoforms 1, 2, 3, 4, and 5 | May differ in size; their relative masses and separation on a blot are not supplied |
| Situation | Likely cause | Next action |
|---|---|---|
| No band in lysate | TGFA can be secreted into the extracellular space | Check conditioned medium alongside a validated positive-control lysate |
| Band higher than expected | Identity or mobility of the higher band is unestablished; Asn25 is an N-glycosylation site | Compare with the ~17 kDa control band and verify antibody specificity |
| Band lower than expected | Signal-peptide or propeptide cleavage may produce smaller TGFA species | Compare lysate and conditioned medium with a validated TGFA control |
| Multiple bands | TGFA has five isoforms and undergoes precursor processing, but their band positions are unknown | Use a validated control to identify TGFA-reactive bands |
| Weak or no signal | Secreted TGFA may be scarce in the sampled lysate | Check conditioned medium and include a positive control |
| Fragments below expected size | Precursor processing is possible, but the identity of smaller fragments is unestablished | Verify antibody specificity and compare sample fractions with a positive control |
Experiment evidence
Western blot images showing TGFA band
M01779-1 · TGFA
← swipe / scroll for more images →
Real WB data behind the band-size expectations in this section.
Sample controls for TGFA Western blot
🧪HPA-IHC candidate guidance (verify in WB): For positive controls for TGFA in Western blot, you can use adrenal gland tissue.
Positive control: Adrenal gland (IHC candidate; verify WB)
Negative control: Adipose tissue (IHC candidate; verify WB)
Loading controls: Run GAPDH, β-actin, and a total-protein stain alongside the samples.
⚠️Feasibility: Because TGFA is secreted, whole-tissue lysates may give weak signal; consider an extracellular protein fraction.
HPA tissue expression evidence for TGFA
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 | Medium | Protein (IHC) | HPA → |
| Appendix | glandular cells | Medium | Protein (IHC) | HPA → |
| Breast | glandular cells | Medium | Protein (IHC) | HPA → |
| Caudate | neuronal cells | Medium | Protein (IHC) | HPA → |
| Cerebral cortex | endothelial cells | Medium | 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 → |
| Ovary | ovarian stroma cells | Not detected | Protein (IHC) | HPA → |
| Bone marrow | hematopoietic cells | Low | Protein (IHC) | HPA → |
| Cerebellum | cells in granular layer | Low | Protein (IHC) | HPA → |
| Oral mucosa | squamous epithelial cells | Low | Protein (IHC) | HPA → |
Section 3
Advanced TGFA Western Blot Tips
Deeper troubleshooting and optimisation questions for TGFA, answered from its protein features.
How should TGFA 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 TGFA isoforms could affect antibody detection?
Isoforms · Five isoforms are listed. Residue 32 is missing in isoforms 2, 3, and 5. At canonical positions 159–160, VV becomes ATLG in isoform 3 or GCRLY in isoforms 4 and 5. Check whether the antibody epitope overlaps these regions when comparing isoform signals.
What does TGFA glycosylation imply for band interpretation?
PTM · UniProt lists one N-linked glycosylation site at Asn25, using canonical sequence numbering. It lies within the annotated propeptide at 24–39. Check which form the antibody detects before attributing a band to glycosylation; the site alone does not establish a visible shift.
Does this guide establish induction of TGFA?
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 TGFA?
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 M01779-1 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.
What matters when quantifying TGFA across samples?
Quantitation · Use the same antibody and conditions across samples, and establish which TGFA form its epitope detects. TGFA has annotated processing regions, five isoforms, and a secreted transforming growth factor alpha form, so signals from different forms should not automatically be combined.
Does the observed TGFA band differ from its predicted mass?
Interpretation · The supplied apparent band and predicted mass are both about 17 kDa. The listed features do not establish a mass shift.
TGFA has a signal peptide at residues 1–23 and a propeptide at 24–39. When interpreting bands, consider whether the antibody detects the precursor or a processed form; these features alone do not predict where either will migrate.
UniProt lists three disulfide bonds. If bands differ between reducing and nonreducing conditions, consider disulfide dependent migration when interpreting them. The feature count alone does not predict a particular band or shift.
Compare them with the annotated signal peptide at 1–23, propeptide at 24–39, Asn25 glycosylation site, and five isoforms. These features offer possible interpretations but do not identify an unexpected band by themselves.
TGFA has a signal peptide at residues 1–23 and a propeptide at 24–39. When interpreting bands, consider whether the antibody detects the precursor or a processed form; these features alone do not predict where either will migrate.
UniProt lists three disulfide bonds. If bands differ between reducing and nonreducing conditions, consider disulfide dependent migration when interpreting them. The feature count alone does not predict a particular band or shift.
Compare them with the annotated signal peptide at 1–23, propeptide at 24–39, Asn25 glycosylation site, and five isoforms. These features offer possible interpretations but do not identify an unexpected band by themselves.
Boster reagents
TGFA 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-TGF alpha Monoclonal Antibody
Cat # M01779-1
Real WB data
Anti-TGF alpha/TGFA Antibody Picoband®
Cat # PA1360
Two the supplier anti-TGFA antibodies have Western blot images for human samples. M01779-1 shows a 17 kDa band in HepG2 and A431 whole cell lysates; PA1360 shows a 6 kDa band with recombinant human TGF alpha. No publication evidence was supplied.
Which to pick: For human cell lysates, choose M01779-1 based on its HepG2 and A431 blot at 1:500. For recombinant human TGF alpha, PA1360 has a blot at 0.5 µg/mL. The images cover different sample types and band sizes.
Source: BosterBio TGFA gene-info card — filtered to Western-blot-capable antibodies; each card shows that product's actual WB validation figure.
References
- UniProt Consortium. UniProt entry P01135.
- Human Protein Atlas. TGFA tissue expression.
- PMC7982073 — target-verified WB comparison
- PMC6006480 — target-verified WB comparison
- PMC5342377 — target-verified WB comparison