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- Table of Contents
GSTM3 / Glutathione S-transferase Mu 3 · Western blot design guide
Design a Western Blot for GSTM3
Real validated GSTM3 Western blot protocols, expected-band and isoform facts, troubleshooting for weak or shifted signal, and recommended anti-GSTM3 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
GSTM3 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 | ~26.6 kDa | |
| Observed band | ~27 kDa | |
| Gel | 12% (catalog A03298-1) | |
| Positive control | Kidney (IHC candidate; verify WB) +3 more | |
| Negative control | Adipose tissue (IHC candidate; verify WB) |
Important caveats
Reasons your observed band may differ from the expected size.
| PTM | Ubl conjugation | |
| Caveat | Modification-state controls | |
| Gene-set association | MSigDB Hallmark membership | |
| Isoform | 1 isoform(s) |
Section 1
Real Curated GSTM3 Western Blot Protocols
The A03298-1 protocol combines labelled catalog values with standard starting conditions. Published comparisons retain their own sample, reagent and detection scope.
| Sample / lysate | human Hela, human MCF-7, human SiHa (catalog A03298-1) |
| Gel % | 12% (catalog A03298-1) |
| Load | 30 ug; reducing conditions (catalog A03298-1) |
| Transfer | a nitrocellulose membrane at 150 mA for 50-90 minutes (catalog A03298-1) |
| Membrane | nitrocellulose membrane (catalog A03298-1) |
| Blocking | 5% non-fat milk/TBS for 1.5 hour at RT (catalog A03298-1) |
| Primary antibody | A03298-1 · 0.5 μg/mL (catalog A03298-1) |
| Primary incubation | overnight at 4°C (catalog A03298-1) |
| Secondary antibody | goat anti-rabbit IgG-HRP, 1:5000 (catalog A03298-1) |
| Secondary incubation | 1.5 hour at RT (catalog A03298-1) |
| Wash | TBS-0.1%Tween 3 times with 5 minutes each (catalog A03298-1) |
| Detection | ECL (catalog A03298-1) |
Section 2
What Is the Expected GSTM3 Western Blot Band Size?
GSTM3 is predicted at 26.6 kDa and observed near 27 kDa under reducing conditions; the small difference has no established cause in the supplied evidence.
| Single band near 27 kDa | GSTM3 monomer, consistent with the empirical reducing-condition band |
| Band near 53 kDa | possible retained GSTM3 homodimer; confirm its identity |
| Bands near 27 and 53 kDa | possible monomer and retained homodimer; confirm both bands |
| 27 kDa band with no 53 kDa band | consistent with dissociation of the homodimer during SDS-PAGE |
💡Expected GSTM3 appearanceUniProt predicts a 26.6 kDa monomer, and antibody QC shows a band at approximately 27 kDa under reducing conditions; confirm band identity with appropriate controls.
| Predicted monomer mass | 26.6 kDa from UniProt, consistent with the observed band near 27 kDa |
| 225-residue protein sequence | underlies the predicted 26.6 kDa monomer mass |
| Homodimer formation | could produce a band near twice the monomer mass if the dimer survives electrophoresis |
| Homodimer dissociation during SDS-PAGE | favors the monomer band near 27 kDa |
| Situation | Likely cause | Next action |
|---|---|---|
| No band in lysate | cytoplasmic GSTM3 may be below detection in the loaded sample | check lysate loading and include a GSTM3-positive control |
| Band higher than expected | a GSTM3 homodimer may have survived electrophoresis | compare denaturing conditions and verify band identity |
| Band lower than expected | identity or sample integrity is uncertain; no cleavage feature is listed | check sample handling and verify the band with an independent antibody |
| Multiple bands | possible monomer and retained homodimer, or nonspecific signals | compare denaturing conditions and use a GSTM3-specific control |
| Weak or no signal | insufficient detectable cytoplasmic GSTM3 or antibody signal | check loading, transfer, and a positive-control lysate |
Experiment evidence
Western blot images showing GSTM3 band
A03298-1 · GSTM3
← swipe / scroll for more images →
Real WB data behind the band-size expectations in this section.
Sample controls for GSTM3 Western blot
🧪HPA-IHC candidate guidance (verify in WB): For positive controls for GSTM3 in Western blot, you can use kidney lysate, which shows high HPA expression.
Positive control: Kidney (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: Cytoplasmic localization and an HPA not-detected tissue make lysate controls feasible.
HPA tissue expression evidence for GSTM3
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 |
|---|---|---|---|---|
| Kidney | distal tubules | High | Protein (IHC) | HPA → |
| Epididymis | glandular cells | Medium | Protein (IHC) | HPA → |
| Seminal vesicle | glandular cells | Medium | Protein (IHC) | HPA → |
| Testis | elongated or late spermatids | 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 → |
| Adrenal gland | glandular cells | 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 → |
Section 3
Advanced GSTM3 Western Blot Tips
Deeper troubleshooting and optimisation questions for GSTM3, answered from its protein features.
How should GSTM3 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 a second band represent a GSTM3 isoform?
Isoforms · The supplied record lists one isoform and no alternative sequence. These features do not support assigning a second band to a documented GSTM3 isoform.
Do GSTM3 modifications explain a shifted band?
PTM · The record includes Ubl conjugation and isopeptide bond keywords but lists no modified-residue coordinates. Those keywords alone do not establish a visible shift or identify a particular shifted band.
Does this guide establish induction of GSTM3?
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 GSTM3?
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 A03298-1 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.
Which band should be quantified for GSTM3?
Quantitation · Quantify a consistently identified band near the observed 27 kDa position. The predicted mass is 26.6 kDa; the supplied features do not establish that other bands are GSTM3.
How does the observed band compare with GSTM3’s predicted mass?
Interpretation · The observed band is approximately 27 kDa, close to the predicted 26.6 kDa for the 225-residue protein. This small difference does not require a band-shift explanation.
GSTM3 is annotated as a homodimer. If a higher band appears, consider that property during interpretation, but the annotation alone does not show that the band is a dimer.
Use the approximately 27 kDa observed band and 26.6 kDa predicted mass as reference points. The record lists one isoform and identifies GSTM3 as a homodimer, but neither feature alone assigns an unexpected band. Its cytoplasmic location may also help assess whether the sample fraction is appropriate.
GSTM3 is annotated as a homodimer. If a higher band appears, consider that property during interpretation, but the annotation alone does not show that the band is a dimer.
Use the approximately 27 kDa observed band and 26.6 kDa predicted mass as reference points. The record lists one isoform and identifies GSTM3 as a homodimer, but neither feature alone assigns an unexpected band. Its cytoplasmic location may also help assess whether the sample fraction is appropriate.
Boster reagents
GSTM3 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-GSTM3 Antibody Picoband®
Cat # A03298-1
The catalog reports one anti-GSTM3 antibody for Western blot, A03298-1, with reported human and mouse reactivity. Its product blot reports a band at approximately 27 kDa using human cell and mouse testis lysates. No independent publication evidence is supplied.
Which to pick: A03298-1 is the only listed option and has a WB image. The documented samples are HeLa, MCF-7, SiHa and THP-1 whole-cell lysates, plus mouse testis lysate; these examples do not establish validation across all human or mouse samples.
Source: BosterBio GSTM3 gene-info card — filtered to Western-blot-capable antibodies; each card shows that product's actual WB validation figure.
References
- UniProt Consortium. UniProt entry P21266.
- Human Protein Atlas. GSTM3 tissue expression.
- PMC7466085 — target-verified WB comparison
- PMC7485563 — target-verified WB comparison
- PMC10912431 — target-verified WB comparison