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- Table of Contents
GPT2 / Alanine aminotransferase 2 · Western blot design guide
Design a Western Blot for GPT2
Source-linked GPT2 Western blot protocol options, expected-band and isoform facts, troubleshooting for weak or shifted signal, and recommended anti-GPT2 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
GPT2 Western Blot Experimental Design Guide
Expected bands, source-linked protocol options, controls and antibodies — the at-a-glance facts below, then the full design guide.
Must know before running
| Expected band | ~57.9 kDa | |
| Observed band | ~58 kDa | |
| Gel | 10% (catalog A07547-1) | |
| Positive control | Colon (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 | Acetylated | |
| Caveat | Modification-state controls | |
| Gene-set association | MSigDB C7 membership | |
| Isoform | 2 isoform(s) |
Section 1
Source-Linked GPT2 Western Blot Protocol Options
The A07547-1 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, human 293T, rat kidney, rat liver, mouse kidney, mouse liver (catalog A07547-1) |
| Gel % | 10% (catalog A07547-1) |
| Load | 30 ug; reducing conditions (catalog A07547-1) |
| Transfer | a nitrocellulose membrane at 150 mA for 50-90 minutes (catalog A07547-1) |
| Membrane | nitrocellulose membrane (catalog A07547-1) |
| Blocking | 5% non-fat milk/TBS for 1.5 hour at RT (catalog A07547-1) |
| Primary antibody | A07547-1 · 0.5 μg/mL (catalog A07547-1) |
| Primary incubation | overnight at 4°C (catalog A07547-1) |
| Secondary antibody | goat anti-rabbit IgG-HRP, 1:5000 (catalog A07547-1) |
| Secondary incubation | 1.5 hour at RT (catalog A07547-1) |
| Wash | TBS-0.1%Tween 3 times with 5 minutes each (catalog A07547-1) |
| Detection | ECL (catalog A07547-1) |
Section 2
What Is the Expected GPT2 Western Blot Band Size?
GPT2 is predicted at 57.9 kDa and observed at ~58 kDa; the supplied evidence does not establish a cause for the small difference.
| Band at ~58 kDa | Matches the empirical GPT2 band and its 57.9 kDa predicted mass |
| Band near ~116 kDa | Could reflect retained GPT2 homodimer; confirm its identity |
| Two bands at different positions | Could reflect isoforms 1 and 2; their migration has not been established |
| One band without an isoform split | Isoforms 1 and 2 may not resolve into distinct bands |
💡Expected GPT2 appearanceGPT2 has a predicted mass of 57.9 kDa and an observed band at ~58 kDa under reducing conditions; confirm band identity with appropriate controls.
| Predicted GPT2 mass | Places the monomer near 57.9 kDa, consistent with the observed ~58 kDa band |
| GPT2 homodimer | Could appear near twice the monomer mass if the dimer remains intact during electrophoresis |
| Isoform 1 | May differ in size from isoform 2, but its mass and migration are not supplied |
| Isoform 2 | May differ in size from isoform 1, but its mass and migration are not supplied |
| Situation | Likely cause | Next action |
|---|---|---|
| Band higher than expected | A retained homodimer or an unrelated band | Check sample preparation and confirm identity with an independent antibody or GPT2 depletion |
| Band lower than expected | An unverified isoform or unrelated band | Confirm identity with GPT2 depletion and an antibody targeting another region |
| Multiple bands | Possible isoforms 1 and 2 or nonspecific binding | Confirm each band with GPT2 depletion and an independent antibody |
| Weak or no signal | Low GPT2 abundance or insufficient assay sensitivity | Check loading and use a GPT2 positive control |
| Fragments below expected size | Possible sample degradation; fragment masses are not supplied | Prepare fresh samples with protease inhibitors and confirm fragment identity |
Experiment evidence
Western blot images showing GPT2 band
A07547-1 · GPT2
← swipe / scroll for more images →
Real WB data behind the band-size expectations in this section.
Sample controls for GPT2 Western blot
🧪HPA-IHC candidate guidance (verify in WB): For positive controls for GPT2 in Western blot, you can use colon tissue, which HPA rates high for GPT2 expression.
Positive control: Colon (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: HPA reports no detection in adipose tissue, but confirm the negative control in your lysate.
HPA tissue expression evidence for GPT2
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 |
|---|---|---|---|---|
| Colon | glandular cells | High | Protein (IHC) | HPA → |
| Duodenum | glandular cells | High | Protein (IHC) | HPA → |
| Kidney | cells in tubules | High | Protein (IHC) | HPA → |
| Small intestine | glandular cells | 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 → |
| Bone marrow | hematopoietic cells | Not detected | Protein (IHC) | HPA → |
| Breast | adipocytes | Not detected | Protein (IHC) | HPA → |
| Cervix | glandular cells | Not detected | Protein (IHC) | HPA → |
| Liver | cholangiocytes | Not detected | Protein (IHC) | HPA → |
Section 3
Advanced GPT2 Western Blot Tips
Deeper troubleshooting and optimisation questions for GPT2, answered from its protein features.
How should GPT2 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 GPT2 isoforms produce different bands?
Isoforms · UniProt lists isoforms 1 and 2. Isoform 2 lacks residues 1–100 of the canonical sequence, so it may run below full-length GPT2. The supplied features do not establish its apparent band position.
Isoform 2 lacks canonical residues 1–100. An antibody whose epitope lies in that region could detect isoform 1 while missing isoform 2. Check the antibody's mapped epitope before interpreting a missing isoform band.
If separate bands are resolved and verified, quantify each isoform consistently. Because isoform 2 lacks canonical residues 1–100, confirm that the antibody detects both isoforms before comparing or combining their signals.
Isoform 2 lacks canonical residues 1–100. An antibody whose epitope lies in that region could detect isoform 1 while missing isoform 2. Check the antibody's mapped epitope before interpreting a missing isoform band.
If separate bands are resolved and verified, quantify each isoform consistently. Because isoform 2 lacks canonical residues 1–100, confirm that the antibody detects both isoforms before comparing or combining their signals.
Which GPT2 modifications matter when interpreting bands?
PTM · In UniProt canonical numbering, lysine 341 carries pyridoxal phosphate, while lysines 415, 505, and 512 are acetylated. These sites do not, by themselves, establish a visible Western-blot shift. Check the numbering convention before comparing them with antibody or paper annotations.
The supplied UniProt features list no glycosylation sites, signal peptide, or propeptide. They therefore provide no basis for assigning an unexpected band to glycosylation or precursor cleavage. Verify band identity before attributing it to a GPT2 modification.
The supplied UniProt features list no glycosylation sites, signal peptide, or propeptide. They therefore provide no basis for assigning an unexpected band to glycosylation or precursor cleavage. Verify band identity before attributing it to a GPT2 modification.
Does this guide establish induction of GPT2?
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 GPT2?
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 A07547-1 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 GPT2 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.
How does GPT2's observed band compare with its predicted mass?
Interpretation · The reported band near 58 kDa closely matches the 57.9 kDa predicted mass. The listed modifications alone do not establish a visible shift or explain a mass difference.
UniProt describes GPT2 as a homodimer, so dimer association is one possibility to investigate. That feature does not establish that a dimer survives the Western-blot conditions or identify a particular higher band.
UniProt describes GPT2 as a homodimer, so dimer association is one possibility to investigate. That feature does not establish that a dimer survives the Western-blot conditions or identify a particular higher band.
Boster reagents
GPT2 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-GPT2 Antibody Picoband®
Cat # A07547-1
The catalog reports one anti-GPT2 antibody, A07547-1, for Western blotting. Its validation image shows an approximately 58 kDa band in human cell, rat tissue, and mouse tissue lysates. The supplied evidence covers these samples under the reported conditions.
Which to pick: A07547-1 is the only listed option. Its WB image includes human HepG2, A431, and 293T cells, plus rat and mouse kidney and liver tissues; choose it when those reported contexts are relevant to your experiment.
Source: BosterBio GPT2 gene-info card — filtered to Western-blot-capable antibodies; each card shows that product's actual WB validation figure.
References
- UniProt Consortium. UniProt entry Q8TD30.
- Human Protein Atlas. GPT2 tissue expression.
- PMC8889437 — target-verified WB comparison
- PMC10363803 — target-verified WB comparison
- PMC12664720 — target-verified WB comparison