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- Table of Contents
PRDX2 / Peroxiredoxin-2 · Western blot design guide
Design a Western Blot for PRDX2
Real validated PRDX2 Western blot protocols, expected-band and isoform facts, troubleshooting for weak or shifted signal, and recommended anti-PRDX2 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
PRDX2 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 | ~21.9 kDa | |
| Observed band | ~22 kDa | |
| Gel | 5–20% (catalog A01982-2) | |
| Positive control | Adrenal gland (IHC candidate; verify WB) +4 more | |
| Negative control | Oral mucosa (IHC candidate; verify WB) |
Important caveats
Reasons your observed band may differ from the expected size.
| PTM | Phosphorylated + Acetylated | |
| Caveat | Redox-state controls | |
| Gene-set association | MSigDB Hallmark membership | |
| Isoform | 2 isoform(s) |
Section 1
Real Curated PRDX2 Western Blot Protocols
The A01982-2 protocol combines labelled catalog values with standard starting conditions. Published comparisons retain their own sample, reagent and detection scope.
| Sample / lysate | human HepG2, human Hela, human 293T (catalog A01982-2) |
| Gel % | 5–20% (catalog A01982-2) |
| Load | 30 ug; reducing conditions (catalog A01982-2) |
| Transfer | a nitrocellulose membrane at 150 mA for 50-90 minutes (catalog A01982-2) |
| Membrane | nitrocellulose membrane (catalog A01982-2) |
| Blocking | 5% non-fat milk/TBS for 1.5 hour at RT (catalog A01982-2) |
| Primary antibody | A01982-2 · 0.5 μg/mL (catalog A01982-2) |
| Primary incubation | overnight at 4°C (catalog A01982-2) |
| Secondary antibody | goat anti-rabbit IgG-HRP, 1:5000 (catalog A01982-2) |
| Secondary incubation | 1.5 hour at RT (catalog A01982-2) |
| Wash | TBS-0.1%Tween 3 times with 5 minutes each (catalog A01982-2) |
| Detection | ECL (catalog A01982-2) |
Section 2
What Is the Expected PRDX2 Western Blot Band Size?
PRDX2 is predicted at 21.9 kDa and observed near 22 kDa; the cause of the small difference is not established.
| Band near 22 kDa | Empirical PRDX2 band, consistent with the 21.9 kDa predicted monomer |
| Band near 44 kDa under nonreducing conditions | Possible oxidation-dependent, disulfide-linked PRDX2 homodimer |
| Bands near 22 and 44 kDa | Possible mixture of monomer and disulfide-linked homodimer |
| Additional bands at different sizes | Could reflect isoforms 1 and 2; distinct migration is unverified |
💡Expected PRDX2 appearanceA ~22 kDa band was observed in reducing whole-cell lysates, close to the 21.9 kDa predicted monomer; confirm band identity with appropriate controls.
| Predicted monomer mass | 21.9 kDa predicted; the observed reducing-condition band is ~22 kDa |
| Disulfide at residue 51 | Can contribute to an oxidation-dependent homodimer near twice the monomer mass |
| Disulfide at residue 172 | Can contribute to an oxidation-dependent homodimer near twice the monomer mass |
| Isoforms 1 and 2 | May differ in size, but their individual masses and migration are unspecified |
| Situation | Likely cause | Next action |
|---|---|---|
| No band in lysate | Cytoplasmic PRDX2 may be below detection or poorly recovered | Check lysate loading and a PRDX2-positive control |
| Band higher than expected | Oxidation-dependent disulfide-linked homodimer or incomplete reduction | Compare reducing and nonreducing lanes |
| Band lower than expected | Identity of the smaller band is unestablished | Check antibody specificity with PRDX2 depletion or another antibody |
| Multiple bands | Monomer and disulfide-linked homodimer may coexist; isoform migration is unverified | Compare reducing and nonreducing lanes and verify band identity |
| Weak or no signal | Insufficient detectable PRDX2 or weak antibody detection | Check loading, transfer, and a PRDX2-positive control |
| Fragments below expected size | Fragment identity is unestablished | Check sample integrity and verify bands with another PRDX2 antibody |
Experiment evidence
Western blot images showing PRDX2 band
A01982-2 · PRDX2
PA1834 · PRDX2
M01982 · PRDX2
← swipe / scroll for more images →
Real WB data behind the band-size expectations in this section.
Sample controls for PRDX2 Western blot
🧪HPA-IHC candidate guidance (verify in WB): For positive controls for PRDX2 in Western blot, you can use adrenal gland tissue, which HPA scores as high.
Positive control: Adrenal gland (IHC candidate; verify WB)
Negative control: Oral mucosa (IHC candidate; verify WB)
Loading controls: Run GAPDH, β-actin, and a total-protein stain alongside the samples.
⚠️Feasibility: Cytoplasmic PRDX2 can be assessed in tissue lysates, with HPA reporting no detection in oral mucosa.
HPA tissue expression evidence for PRDX2
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 | High | Protein (IHC) | HPA → |
| Bone marrow | hematopoietic cells | High | Protein (IHC) | HPA → |
| Caudate | neuronal cells | High | Protein (IHC) | HPA → |
| Cerebellum | cells in molecular layer | High | Protein (IHC) | HPA → |
| Cerebral cortex | neuronal cells | High | Protein (IHC) | HPA → |
Lower expression tissues · IHC evidence, not confirmed WB-negative controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| Oral mucosa | squamous epithelial cells | Not detected | Protein (IHC) | HPA → |
| Vagina | squamous epithelial cells | Not detected | Protein (IHC) | HPA → |
| Cervix | glandular cells | Low | Protein (IHC) | HPA → |
| Adipose tissue | adipocytes | Medium | Protein (IHC) | HPA → |
| Appendix | glandular cells | Medium | Protein (IHC) | HPA → |
Section 3
Advanced PRDX2 Western Blot Tips
Deeper troubleshooting and optimisation questions for PRDX2, answered from its protein features.
How should PRDX2 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 PRDX2 isoforms produce different bands?
Isoforms · Two isoforms are listed. Isoform 2 replaces canonical residues 87–198 with a different sequence, so an antibody targeting that region may recognize the isoforms differently. The supplied features do not establish where isoform 2 migrates.
Isoform 2 replaces canonical residues 87–198. Therefore, canonical sites 112, 182, and 196 should not be assigned the same coordinates or modifications in isoform 2 without sequence mapping. The listed N-acetylalanine at position 2 lies outside the replacement.
Isoform 2 replaces canonical residues 87–198. Therefore, canonical sites 112, 182, and 196 should not be assigned the same coordinates or modifications in isoform 2 without sequence mapping. The listed N-acetylalanine at position 2 lies outside the replacement.
Which PRDX2 modifications could affect band interpretation?
PTM · In UniProt canonical coordinates, PRDX2 has N-acetylalanine 2, phosphoserine 112, phosphothreonine 182, and N6-acetyllysine 196. These sites identify possible sources of molecular variation, but their presence does not establish a visible band shift. Check the numbering convention when comparing antibody or paper annotations.
Does this guide establish induction of PRDX2?
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 PRDX2?
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 A01982-2 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 PRDX2 bands be quantified across samples?
Quantitation · Use consistent reducing conditions when comparing the approximately 22 kDa band across samples: PRDX2 can form disulfide-linked dimers upon oxidation. If higher bands are measured, report them separately and identify them before combining their signal with the 22 kDa band.
Why is the PRDX2 band near 22 kDa?
Interpretation · The observed band at about 22 kDa is close to the predicted 21.9 kDa for canonical PRDX2. The listed modifications alone do not establish a visible shift or explain a mass difference.
PRDX2 can form a disulfide-linked homodimer upon oxidation, and five homodimers can assemble into a decamer. A higher band under nonreducing conditions may therefore warrant comparison with a reducing sample. The features do not establish the identity of any particular higher band.
Compare reducing and nonreducing samples to assess whether a higher band is consistent with oxidation-linked dimerization. Also check whether the antibody recognizes the region replaced in isoform 2 (canonical residues 87–198). Neither feature alone identifies an unexpected band.
PRDX2 can form a disulfide-linked homodimer upon oxidation, and five homodimers can assemble into a decamer. A higher band under nonreducing conditions may therefore warrant comparison with a reducing sample. The features do not establish the identity of any particular higher band.
Compare reducing and nonreducing samples to assess whether a higher band is consistent with oxidation-linked dimerization. Also check whether the antibody recognizes the region replaced in isoform 2 (canonical residues 87–198). Neither feature alone identifies an unexpected band.
Boster reagents
PRDX2 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-peroxiredoxin 2/PRDX2 Antibody Picoband®
Cat # A01982-2
Real WB data
Anti-Peroxiredoxin 2/PRDX2 Antibody Picoband®
Cat # PA1834
Real WB data
Anti-Peroxiredoxin 2 Rabbit Monoclonal Antibody
Cat # M01982
Three the supplier anti-PRDX2 antibodies have Western blot images. A01982-2 shows human cell lysates; PA1834 shows rat tissues and human cell lysates; M01982 shows human, rat, and mouse cell lysates. Approximately 22 kDa bands are reported for A01982-2 and M01982.
Which to pick: For human samples, all three have pictured WB examples. For rat tissue, PA1834 shows brain and kidney lysates; M01982 shows rat RH35 cells. For mouse samples, M01982 shows Neuro-2a lysate, while PA1834 lists mouse reactivity without a mouse lane in its caption.
Source: BosterBio PRDX2 gene-info card — filtered to Western-blot-capable antibodies; each card shows that product's actual WB validation figure.
References
- UniProt Consortium. UniProt entry P32119.
- Human Protein Atlas. PRDX2 tissue expression.
- PMC11375325 — target-verified WB comparison
- PMC6811995 — target-verified WB comparison
- PMC6339520 — target-verified WB comparison