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- Table of Contents
GPX2 / Glutathione peroxidase 2 · Western blot design guide
Design a Western Blot for GPX2
Real validated GPX2 Western blot protocols, expected-band and isoform facts, troubleshooting for weak or shifted signal, and recommended anti-GPX2 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
GPX2 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 | ~22 kDa | |
| Observed band | ~22 kDa | |
| Gel | 12% (catalog M03837) | |
| Positive control | Duodenum (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 | — | |
| Caveat | Reducing conditions | |
| Gene-set association | MSigDB Hallmark membership | |
| Isoform | 1 isoform(s) |
Section 1
Real Curated GPX2 Western Blot Protocols
The M03837 protocol combines labelled catalog values with standard starting conditions. Published comparisons retain their own sample, reagent and detection scope.
| Sample / lysate | human OE19, human A549, human HepG2 (catalog M03837) |
| Gel % | 12% (catalog M03837) |
| Load | 30 ug; reducing conditions (catalog M03837) |
| Transfer | a nitrocellulose membrane at 150 mA for 50-90 minutes (catalog M03837) |
| Membrane | nitrocellulose membrane (catalog M03837) |
| Blocking | 5% non-fat milk/TBS for 1.5 hour at RT (catalog M03837) |
| Primary antibody | M03837 · 1:500 (catalog M03837) |
| Primary incubation | overnight at 4°C (catalog M03837) |
| Secondary antibody | goat anti-rabbit IgG-HRP, 1:5000 (catalog M03837) |
| Secondary incubation | 1.5 hour at RT (catalog M03837) |
| Wash | TBS-0.1%Tween 3 times with 5 minutes each (catalog M03837) |
| Detection | ECL (catalog M03837) |
Section 2
What Is the Expected GPX2 Western Blot Band Size?
GPX2 is predicted at 22 kDa and observed at ~22 kDa in reducing whole-cell blots; the supplied evidence establishes no size difference.
| Band at ~22 kDa | Matches the empirical GPX2 band and predicted monomer mass |
| Single sharp band near 22 kDa | Consistent with the one listed GPX2 isoform |
| ~22 kDa band in whole-cell lysate | Consistent with cytosolic GPX2 |
| Higher molecular weight band after incomplete denaturation | Could reflect retained GPX2 homotetramer; identity needs confirmation |
💡Expected GPX2 appearanceGPX2 has a predicted monomer mass of 22 kDa and an observed band at ~22 kDa in reducing whole-cell blots; confirm identity with appropriate positive and negative controls.
| UniProt predicted monomer mass | Places the expected monomer band near the observed ~22 kDa |
| Calculated sequence mass of 21,954 Da | Rounds to approximately 22 kDa |
| 190-residue GPX2 chain | Provides the sequence underlying the predicted 22 kDa mass |
| Homotetramer | Could appear above the monomer if the complex persists during electrophoresis; no such band is established here |
| Situation | Likely cause | Next action |
|---|---|---|
| No band in lysate | Cytosolic GPX2 may be below detection in the tested lysate | Check loading and antibody performance with a GPX2-positive lysate |
| Band higher than expected | Incomplete denaturation could retain the GPX2 homotetramer, or the band could be nonspecific | Check denaturation and confirm identity with an independent antibody or GPX2 depletion |
| Band lower than expected | Possible proteolysis or nonspecific binding; no cleavage feature is listed | Prepare fresh lysate with protease inhibitors and confirm band identity |
| Multiple bands | Possible incomplete denaturation, proteolysis, or nonspecific binding | Compare fully denatured samples and use GPX2 depletion to identify specific bands |
| Weak or no signal | GPX2 abundance or assay sensitivity may be insufficient | Verify loading and transfer, then compare with a GPX2-positive lysate |
Experiment evidence
Western blot images showing GPX2 band
M03837 · GPX2
M03837 · GPX2
← swipe / scroll for more images →
Real WB data behind the band-size expectations in this section.
Sample controls for GPX2 Western blot
🧪HPA-IHC candidate guidance (verify in WB): For positive controls for GPX2 in Western blot, you can use duodenum lysate, which shows high expression in HPA.
Positive control: Duodenum (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: Cytosolic GPX2 should be accessible in tissue lysates, with adipose tissue providing an HPA not-detected comparison.
HPA tissue expression evidence for GPX2
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 |
|---|---|---|---|---|
| Duodenum | paneth cells | High | Protein (IHC) | HPA → |
| Kidney | distal tubules | High | Protein (IHC) | HPA → |
| Small intestine | paneth cells | High | Protein (IHC) | HPA → |
| Urinary bladder | urothelial cells | High | Protein (IHC) | HPA → |
| Appendix | enterocytes | 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 → |
| 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 GPX2 Western Blot Tips
Deeper troubleshooting and optimisation questions for GPX2, answered from its protein features.
Where should the GPX2 band appear?
Band shift · The predicted mass and observed band are both about 22 kDa. Identify the main GPX2 band near that position, while allowing for small differences in apparent migration.
Can GPX2 isoforms explain additional bands?
Isoforms · The supplied UniProt record lists one isoform and no alternative sequence. It does not support assigning additional bands to GPX2 isoforms.
Do listed modifications explain a GPX2 band shift?
PTM · No modified residues or glycosylation sites are listed. The sequence contains selenocysteine, but its presence alone does not establish a visible shift or explain a mass difference.
Does this guide establish induction of GPX2?
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.
What transfer method to use for GPX2 Western blot?
Transfer · Optimize transfer for a protein near 22 kDa and verify that the band is retained on the membrane. The supplied features do not establish a preference for a particular transfer method.
How should blocking be optimized?
Blocking · Standard workflow guidance: follow the M03837 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 GPX2 bands be quantified?
Quantitation · Quantify the band near 22 kDa using consistent sample preparation and normalization across lanes. GPX2 is listed as cytosolic, so keep the sampled fraction consistent when comparing abundance.
How should unexpected high-mass GPX2 bands be interpreted?
Interpretation · GPX2 is reported as a homotetramer, but that feature alone does not identify a high-mass band. Compare denaturing conditions and band behavior before attributing an additional band to GPX2 oligomers.
Boster reagents
GPX2 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-Glutathione Peroxidase 2 Rabbit Monoclonal Antibody
Cat # M03837
M03837 is a rabbit monoclonal anti-GPX2 antibody listed for human samples. Its Western blot image reports a band near the expected 22 kDa in OE19, A549, HepG2, and RT4 whole cell lysates. Evidence beyond these tested samples is not supplied.
Which to pick: M03837 is the only listed GPX2 antibody. For human Western blots, it has a reported validation image using four human cell lysates at a 1:500 primary dilution. Those examples do not establish performance in every human sample.
Source: BosterBio GPX2 gene-info card — filtered to Western-blot-capable antibodies; each card shows that product's actual WB validation figure.
References
- UniProt Consortium. UniProt entry P18283.
- Human Protein Atlas. GPX2 tissue expression.
- PMC9313920 — target-verified WB comparison
- PMC6883322 — target-verified WB comparison
- PMC5787487 — target-verified WB comparison