This website uses cookies to ensure you get the best experience on our website.
- Table of Contents
GSTO1 / Glutathione S-transferase omega-1 · Western blot design guide
Design a Western Blot for GSTO1
Source-linked GSTO1 Western blot protocol options, expected-band and isoform facts, troubleshooting for weak or shifted signal, and recommended anti-GSTO1 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
GSTO1 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 | ~27.6 kDa | |
| Observed band | Approximately 28–30 kDa | |
| Gel | 10% (catalog A02461-1) | |
| Positive control | Liver (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 | Modification-state controls | |
| Gene-set association | MSigDB Hallmark membership | |
| Isoform | 3 isoform(s) |
Section 1
Source-Linked GSTO1 Western Blot Protocol Options
The A02461-1 protocol combines labelled catalog values with standard starting conditions. Published comparisons retain their own sample, reagent and detection scope.
| Sample / lysate | human Hela, human Jurkat, human PC-3 (catalog A02461-1) |
| Gel % | 10% (catalog A02461-1) |
| Load | 30 ug; reducing conditions (catalog A02461-1) |
| Transfer | a nitrocellulose membrane at 150 mA for 50-90 minutes (catalog A02461-1) |
| Membrane | nitrocellulose membrane (catalog A02461-1) |
| Blocking | 5% non-fat milk/TBS for 1.5 hour at RT (catalog A02461-1) |
| Primary antibody | A02461-1 · 1:1000 (catalog A02461-1) |
| Primary incubation | overnight at 4°C (catalog A02461-1) |
| Secondary antibody | goat anti-rabbit IgG-HRP, 1:5000 (catalog A02461-1) |
| Secondary incubation | 1.5 hour at RT (catalog A02461-1) |
| Wash | TBS-0.1%Tween 3 times with 5 minutes each (catalog A02461-1) |
| Detection | ECL (catalog A02461-1) |
Section 2
What Is the Expected GSTO1 Western Blot Band Size?
GSTO1 is predicted at 27.6 kDa and observed at approximately 28–30 kDa; the cause of the difference is not established.
| Band at approximately 28–30 kDa | Empirical GSTO1 band in reducing lysate blots; confirm identity with controls |
| Sharp band near 27.6 kDa | Consistent with the predicted GSTO1 monomer mass |
| Band near 55 kDa | Could represent a homodimer if it remains intact during electrophoresis |
| Several distinct bands | Could reflect isoforms 1, 2, and 3; their migration differences are unknown |
💡Expected GSTO1 appearanceGSTO1 has a predicted mass of 27.6 kDa and an empirical band at approximately 28–30 kDa; the cause of the difference is unestablished, so confirm band identity with appropriate controls.
| Predicted GSTO1 mass | Places the monomer near 27.6 kDa; empirical blots show approximately 28–30 kDa |
| Homodimer formation | Could yield a band near twice the monomer mass if the dimer survives electrophoresis |
| Splice isoform 1 | May migrate differently from other isoforms; its individual mass is not supplied |
| Splice isoform 2 | May migrate differently from other isoforms; its individual mass is not supplied |
| Splice isoform 3 | May migrate differently from other isoforms; its individual mass is not supplied |
| Situation | Likely cause | Next action |
|---|---|---|
| Band higher than expected | An intact homodimer is possible, but its survival during electrophoresis is unverified | Compare reducing and nonreducing samples and confirm band identity |
| Band lower than expected | An alternative isoform or sample degradation is possible | Compare with a validated GSTO1 control and repeat with fresh, protease-inhibited lysate |
| Multiple bands | GSTO1 has three splice isoforms, but their band positions are unknown | Check band specificity with GSTO1 depletion or an independent antibody |
| Weak or no signal | GSTO1 signal or antibody detection may be insufficient | Check lysate loading, transfer, and a positive lysate control |
| Fragments below expected size | Proteolysis during sample handling is possible | Prepare fresh lysate with protease inhibitors and compare with a validated control |
Experiment evidence
Western blot images showing GSTO1 band
A02461-1 · GSTO1
A02461-1 · GSTO1
M02461 · GSTO1
M02461-1 · GSTO1
← swipe / scroll for more images →
Real WB data behind the band-size expectations in this section.
Sample controls for GSTO1 Western blot
🧪HPA-IHC candidate guidance (verify in WB): For positive controls for GSTO1 in Western blot, you can use liver lysate, as HPA reports high expression in liver.
Positive control: Liver (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: A cytosolic liver signal is feasible; confirm that GSTO1 is absent from the adipose tissue lysate used as the negative control.
HPA tissue expression evidence for GSTO1
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 |
|---|---|---|---|---|
| Liver | hepatocytes | High | Protein (IHC) | HPA → |
| Soft tissue | chondrocytes | High | Protein (IHC) | HPA → |
| Adrenal gland | glandular cells | Medium | Protein (IHC) | HPA → |
| Caudate | neuronal cells | Medium | Protein (IHC) | HPA → |
| Lung | macrophages | 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 → |
| Appendix | glandular cells | Not detected | Protein (IHC) | HPA → |
| Bone marrow | hematopoietic cells | Not detected | Protein (IHC) | HPA → |
| Breast | adipocytes | Not detected | Protein (IHC) | HPA → |
| Bronchus | respiratory epithelial cells | Not detected | Protein (IHC) | HPA → |
Section 3
Advanced GSTO1 Western Blot Tips
Deeper troubleshooting and optimisation questions for GSTO1, answered from its protein features.
How should GSTO1 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 GSTO1 isoforms produce different bands?
Isoforms · Yes. UniProt lists three isoforms: isoform 3 lacks residues 1–28, and isoform 2 lacks residues 123–155, relative to the canonical sequence. These deletions could change band size, but the features do not establish where either isoform runs.
Check the antibody's stated epitope against the alternative sequences. An epitope within canonical residues 1–28 is absent from isoform 3; one within 123–155 is absent from isoform 2. The supplied features do not identify any antibody epitope.
Check the antibody's stated epitope against the alternative sequences. An epitope within canonical residues 1–28 is absent from isoform 3; one within 123–155 is absent from isoform 2. The supplied features do not identify any antibody epitope.
Which GSTO1 modifications matter when interpreting bands?
PTM · UniProt lists N-acetylserine at residue 2, N6-acetyllysine at 57, 143, 148 and 152, and phosphoserine at 129. These are canonical UniProt coordinates; antibody or paper numbering may differ. Their presence alone does not establish a visible band shift.
Does this guide establish induction of GSTO1?
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 GSTO1?
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 A02461-1 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 GSTO1 bands be quantified across samples?
Quantitation · Quantify the same band consistently across samples and report whether it is the approximately 28–30 kDa band. Because GSTO1 has three listed isoforms, a band measurement cannot automatically be treated as the total amount of every isoform.
Why does GSTO1 run at 28–30 kDa instead of 27.6 kDa?
Interpretation · The reported band is approximately 28–30 kDa, close to the predicted 27.6 kDa. The listed modifications do not establish a visible shift or explain the difference.
UniProt describes GSTO1 as a homodimer, but that annotation alone cannot identify a higher Western blot band. The supplied features do not establish whether a dimer survives the blot conditions.
Consider the two listed sequence deletions when comparing possible isoform sizes: residues 1–28 are missing in isoform 3, and 123–155 are missing in isoform 2. Size alone cannot assign an unexpected band to either isoform.
UniProt describes GSTO1 as a homodimer, but that annotation alone cannot identify a higher Western blot band. The supplied features do not establish whether a dimer survives the blot conditions.
Consider the two listed sequence deletions when comparing possible isoform sizes: residues 1–28 are missing in isoform 3, and 123–155 are missing in isoform 2. Size alone cannot assign an unexpected band to either isoform.
Boster reagents
GSTO1 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-GSTO1 Antibody
Cat # A02461-1
Real WB data
Anti-GSTO1 Rabbit Monoclonal Antibody
Cat # M02461
Real WB data
Anti-GSTO1 Rabbit Monoclonal Antibody
Cat # M02461-1
Three the supplier anti-GSTO1 antibodies have WB images. A02461-1 shows an approximately 28 kDa band in human cell and rat tissue lysates under reported conditions. The monoclonal images show HepG2 or HeLa lysate; the supplied captions provide less experimental detail.
Which to pick: Choose A02461-1 for the most detailed WB example, including human and rat samples; mouse reactivity is listed but no mouse lane is shown. Choose M02461 for its HepG2 example or M02461-1 for its HeLa example. Both are rabbit monoclonals.
Source: BosterBio GSTO1 gene-info card — filtered to Western-blot-capable antibodies; each card shows that product's actual WB validation figure.
References
- UniProt Consortium. UniProt entry P78417.
- Human Protein Atlas. GSTO1 tissue expression.
- PMC6966599 — target-verified WB comparison
- PMC9521632 — target-verified WB comparison
- PMC10471458 — target-verified WB comparison