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- Table of Contents
PLA2G2A / Phospholipase A2, membrane associated · Western blot design guide
Design a Western Blot for PLA2G2A
Source-linked PLA2G2A Western blot protocol options, expected-band and isoform facts, troubleshooting for weak or shifted signal, and recommended anti-PLA2G2A 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
PLA2G2A 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 | ~16.1 kDa | |
| Observed band | ~16 kDa | |
| Gel | 5–20% (catalog M02259-1) | |
| 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 | Cleaved | |
| Caveat | Signal-peptide cleavage | |
| Gene-set association | MSigDB Hallmark membership | |
| Isoform | 1 isoform(s) |
Section 1
Source-Linked PLA2G2A Western Blot Protocol Options
The M02259-1 protocol combines labelled catalog values with standard starting conditions. Published comparisons retain their own sample, reagent and detection scope.
| Sample / lysate | human A549, human HepG2, human RT4, human Hacat, rat small intestine, rat C6, mouse small intestine, mouse EL-4 (catalog M02259-1) |
| Gel % | 5–20% (catalog M02259-1) |
| Load | 30 ug; reducing conditions (catalog M02259-1) |
| Transfer | a nitrocellulose membrane at 150 mA for 50-90 minutes (catalog M02259-1) |
| Membrane | nitrocellulose membrane (catalog M02259-1) |
| Blocking | 5% non-fat milk/TBS for 1.5 hour at RT (catalog M02259-1) |
| Primary antibody | M02259-1 · 1:500 (catalog M02259-1) |
| Primary incubation | overnight at 4°C (catalog M02259-1) |
| Secondary antibody | goat anti-rabbit IgG-HRP, 1:500 (catalog M02259-1) |
| Secondary incubation | 1.5 hour at RT (catalog M02259-1) |
| Wash | TBS-0.1%Tween 3 times with 5 minutes each (catalog M02259-1) |
| Detection | ECL (catalog M02259-1) |
Section 2
What Is the Expected PLA2G2A Western Blot Band Size?
UniProt predicts 16.1 kDa and reducing blots show ~16 kDa; the evidence does not establish a cause for their small difference.
| Band at ~16 kDa | matches the reported PLA2G2A band in reducing lysate blots |
| Band below the precursor | could reflect signal-peptide cleavage; confirm its identity |
| Little or no band in whole-cell lysate | may reflect secretion or sample-dependent abundance |
| Band in a membrane fraction | consistent with peripheral membrane localization |
💡Expected PLA2G2A appearanceUniProt predicts a 16.1 kDa precursor, while reducing lysate blots report a ~16 kDa band; confirm identity with antibody specificity and appropriate sample controls.
| UniProt predicted mass | places the 144-residue precursor near 16.1 kDa |
| 144-residue precursor | provides the full-length reference for comparing a cleaved product |
| Signal peptide at residues 1–20 | cleavage could yield a smaller mature protein, but its migration is not supplied |
| Intrachain disulfide bonds | may affect migration under different reduction conditions without establishing a doubled mass |
| Situation | Likely cause | Next action |
|---|---|---|
| No band in lysate | PLA2G2A can be secreted | check conditioned medium alongside lysate |
| Band higher than expected | identity or reduction-dependent migration is uncertain | compare fully reducing conditions and an antibody specificity control |
| Band lower than expected | signal-peptide cleavage is possible | check antibody epitope coverage and compare precursor and mature controls |
| Multiple bands | precursor processing or unrelated antibody binding may contribute | compare relevant sample fractions and validate band identity |
| Weak or no signal | protein may be scarce in the sampled lysate | test conditioned medium or a suitable positive-control sample |
| Fragments below expected size | sample degradation or processing may be involved | use fresh samples with protease inhibitors and verify band identity |
Experiment evidence
Western blot images showing PLA2G2A band
M02259-1 · PLA2G2A
← swipe / scroll for more images →
Real WB data behind the band-size expectations in this section.
Sample controls for PLA2G2A Western blot
🧪HPA-IHC candidate guidance (verify in WB): For positive controls for PLA2G2A in Western blot, you can use duodenum tissue, which HPA reports as highly positive.
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: PLA2G2A is secreted, so whole-cell lysates may give weak signal; consider an extracellular fraction.
HPA tissue expression evidence for PLA2G2A
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 → |
| Small intestine | paneth cells | High | Protein (IHC) | HPA → |
| Appendix | germinal center cells | Medium | Protein (IHC) | HPA → |
| Colon | enterocytes | Medium | Protein (IHC) | HPA → |
| Prostate | glandular cells | 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 PLA2G2A Western Blot Tips
Deeper troubleshooting and optimisation questions for PLA2G2A, answered from its protein features.
How should PLA2G2A 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.
Are alternative isoforms expected to produce additional PLA2G2A bands?
Isoforms · The supplied record lists one isoform and no alternative sequence. It provides no isoform-based explanation for additional bands; assess those bands separately.
Can annotated modifications explain extra PLA2G2A bands?
PTM · No glycosylation sites or modified residues are listed. The record does list seven disulfide bonds, which make reducing conditions relevant when comparing lanes. These features alone do not establish the cause of an extra band or a visible shift.
Does this guide establish induction of PLA2G2A?
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 PLA2G2A Western blot?
Transfer · PLA2G2A is predicted at 16.1 kDa, so choose transfer conditions and a membrane suitable for retaining small proteins. Check transfer efficiency and avoid conditions that let the band pass through the membrane.
How should blocking be optimized?
Blocking · Standard workflow guidance: follow the M02259-1 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 fractions should be compared when quantifying PLA2G2A?
Quantitation · PLA2G2A is annotated as secreted and as a peripheral protein at the cell and mitochondrial outer membranes. Consider medium and relevant cellular fractions, and account for their different sample inputs when comparing signal.
Does the ~16 kDa band match PLA2G2A's predicted mass?
Interpretation · The observed ~16 kDa band is close to the 16.1 kDa prediction for the 144-residue sequence. PLA2G2A also has a signal peptide at UniProt positions 1–20, so processing could change its mass. Band position alone cannot establish whether the signal peptide was removed.
The annotated signal peptide spans UniProt positions 1–20. Compare the ~16 kDa band with the predicted full-length mass of 16.1 kDa, but do not assign a processed form from migration alone.
Check whether the ~16 kDa band is present across the sampled compartments and compare reducing conditions, given the seven annotated disulfide bonds. The single listed isoform and lack of annotated glycosylation or modified residues do not identify other bands. Avoid assigning them to processing or modification based on size alone.
The annotated signal peptide spans UniProt positions 1–20. Compare the ~16 kDa band with the predicted full-length mass of 16.1 kDa, but do not assign a processed form from migration alone.
Check whether the ~16 kDa band is present across the sampled compartments and compare reducing conditions, given the seven annotated disulfide bonds. The single listed isoform and lack of annotated glycosylation or modified residues do not identify other bands. Avoid assigning them to processing or modification based on size alone.
Boster reagents
PLA2G2A 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-PLA2G2A Rabbit Monoclonal Antibody
Cat # M02259-1
M02259-1 is a rabbit monoclonal anti-PLA2G2A antibody listed for human, mouse, and rat Western blots. Its product image reports a band near the expected 16 kDa in the specified lysates. The supplied evidence is a product blot; independent validation is not provided.
Which to pick: M02259-1 is the only listed option. Its Western blot image includes human cell, rat cell and intestine, and mouse cell and intestine lysates, using a 1:500 primary antibody dilution. Match your planned sample and conditions to those examples.
Source: BosterBio PLA2G2A gene-info card — filtered to Western-blot-capable antibodies; each card shows that product's actual WB validation figure.
References
- UniProt Consortium. UniProt entry P14555.
- Human Protein Atlas. PLA2G2A tissue expression.
- PMC5225502 — target-verified WB comparison
- PMC12998596 — target-verified WB comparison
- PMC4687730 — target-verified WB comparison