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- Table of Contents
PMP22 / Peripheral myelin protein 22 · Western blot design guide
Design a Western Blot for PMP22
Real validated PMP22 Western blot protocols, expected-band and isoform facts, troubleshooting for weak or shifted signal, and recommended anti-PMP22 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
PMP22 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 | ~17.9 kDa | |
| Gel | 15% (standard starting point) | |
| Negative control | Suggested KO / knockdown lysate |
Important caveats
Reasons your observed band may differ from the expected size.
| PTM | Glycosylated + Ubl conjugation | |
| Caveat | Modification-state controls | |
| Gene-set association | MSigDB Hallmark membership | |
| Isoform | 1 isoform(s) |
Section 1
Real Curated PMP22 Western Blot Protocols
The M00890 protocol combines labelled catalog values with standard starting conditions. Published comparisons retain their own sample, reagent and detection scope.
| Sample / lysate | 293T cell lysate (catalog M00890) |
| Gel % | 15% (standard starting point) |
| Load | 20–30 µg total protein per lane; optimize for abundance (standard starting point) |
| Transfer | Short semi-dry transfer; verify retention (standard starting point) |
| Membrane | 0.2 µm PVDF (standard starting point) |
| Blocking | 5% milk or 5% BSA in TBST (standard starting point) |
| Primary antibody | M00890; use the WB datasheet starting dilution (standard starting point) |
| Primary incubation | Overnight at 4 °C (standard starting point) |
| Secondary antibody | Species-matched HRP conjugate at validated dilution (standard starting point) |
| Secondary incubation | 1 h at room temperature (standard starting point) |
| Wash | 3 × 5 min in TBST (standard starting point) |
| Detection | ECL; bracket exposures to avoid saturation (standard starting point) |
Section 2
What Is the Expected PMP22 Western Blot Band Size?
PMP22 is predicted at 17.9 kDa; Asn41 N-linked glycosylation may affect migration, but no empirical band size or visible shift is established.
| Band near 17.9 kDa | Consistent with the predicted full-length PMP22 mass; confirm identity with antibody controls. |
| Band above 17.9 kDa | Could reflect N-linked glycosylation at Asn41; the site alone does not establish a visible shift. |
| Two nearby bands | Could reflect differing Asn41 glycosylation states, but distinct bands are not established. |
| Weak band in soluble lysate | Consistent with PMP22 localization to the cell membrane. |
💡Expected PMP22 appearancePMP22 has a predicted mass of 17.9 kDa, but no empirical band size is supplied; assess bands near that size with antibody controls, since Asn41 glycosylation may affect migration.
| UniProt predicted mass | Sets the 17.9 kDa full-length reference size. |
| Full-length 160-residue sequence | Underlies the predicted mass; no cleavage feature is listed. |
| Asn41 N-linked glycosylation | May increase apparent size if the site carries a glycan. |
| Asn41 glycan occupancy | An unoccupied site would lack that potential glycan-related increase; a separate band is not established. |
| Situation | Likely cause | Next action |
|---|---|---|
| No band in lysate | Membrane-associated PMP22 may be poorly recovered. | Check the membrane fraction and extraction conditions. |
| Band higher than expected | Asn41 N-linked glycosylation is a possible contributor. | Compare untreated and deglycosylated samples with identity controls. |
| Band lower than expected | The supplied features do not establish a smaller PMP22 product. | Verify band identity with a second antibody or a PMP22 control. |
| Multiple bands | Different Asn41 glycosylation states are possible but unproven. | Compare deglycosylated samples and verify band identity. |
| Weak or no signal | Cell membrane localization may limit recovery during extraction. | Check membrane protein recovery and antibody performance. |
Experiment evidence
Western blot images showing PMP22 band
M00890 · PMP22
← swipe / scroll for more images →
Real WB data behind the band-size expectations in this section.
Sample controls for PMP22 Western blot
🧪For positive controls for PMP22 in Western blot, you can use a positive tissue or cell line once HPA evidence identifies one.
Positive control: No high/medium HPA tissue identified
Negative control: Suggested KO / knockdown lysate
Loading controls: Run GAPDH, β-actin, and a total-protein stain alongside the samples.
⚠️Feasibility: HPA provides no tissue controls; as a multi-pass membrane protein, PMP22 may require effective membrane-protein extraction.
HPA tissue expression evidence for PMP22
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 |
|---|---|---|---|---|
| No high/medium HPA tissues identified in the supplied evidence. | ||||
Lower expression tissues · IHC evidence, not confirmed WB-negative controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| No lower-expression tissue rows available in the supplied evidence. | ||||
Section 3
Advanced PMP22 Western Blot Tips
Deeper troubleshooting and optimisation questions for PMP22, answered from its protein features.
How should PMP22 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 annotated PMP22 isoforms explain multiple bands?
Isoforms · The supplied record lists one isoform and no alternative sequence. It therefore provides no annotated isoform masses with which to assign additional bands.
Which PMP22 modification should I consider when interpreting bands?
PTM · UniProt annotates N-linked glycosylation at Asn41, using UniProt sequence numbering. Its keywords also include Ubl conjugation, but no modified residue is listed for that. Do not assign a particular band to either modification without experimental evidence.
Does this guide establish induction of PMP22?
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 PMP22 Western blot?
Transfer · PMP22 is a 17.9 kDa multi-pass membrane protein. Verify that your transfer conditions retain a protein of this size on the membrane, and check the post-transfer gel. The supplied features do not identify a preferred transfer method.
How should I optimize blocking for PMP22?
Blocking · Compare blocking conditions by PMP22 signal and background in your samples. Its membrane location and sequence do not specify a blocker or predict which condition will work best.
How should I quantify PMP22 if several bands appear?
Quantitation · Define which band or bands you will measure before comparing samples, and use the same definition throughout. The record supplies no observed band position; its Asn41 glycosylation annotation alone cannot establish which bands represent PMP22.
Why might PMP22 migrate differently from its predicted mass?
Interpretation · PMP22 has a predicted mass of 17.9 kDa and one annotated N-linked glycosylation site, UniProt Asn41. Glycosylation could affect migration, but no observed band position is supplied, and the annotation alone does not establish a visible shift.
Compare them with the predicted 17.9 kDa mass, while recognizing that no empirical band position is supplied. Asn41 glycosylation is a possible consideration, but the features do not prove a shift or identify extra bands. The record lists no alternative sequence to support an isoform assignment.
Compare them with the predicted 17.9 kDa mass, while recognizing that no empirical band position is supplied. Asn41 glycosylation is a possible consideration, but the features do not prove a shift or identify extra bands. The record lists no alternative sequence to support an isoform assignment.
Boster reagents
PMP22 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-PMP22 Monoclonal Antibody
Cat # M00890
The catalog reports one anti-PMP22 monoclonal antibody, M00890, with stated Human and Rat reactivity. Its Western blot image shows PMP22 expression in 293T cell lysate; the supplied evidence does not show a Rat blot.
Which to pick: M00890 is the only listed option and has a Western blot image from 293T cell lysate. For Rat samples, reactivity is listed, but no Rat Western blot example is supplied.
Source: BosterBio PMP22 gene-info card — filtered to Western-blot-capable antibodies; each card shows that product's actual WB validation figure.
References
- UniProt Consortium. UniProt entry Q01453.
- Human Protein Atlas. PMP22 tissue expression.
- PMC5338570 — target-verified WB comparison
- PMC11784151 — target-verified WB comparison
- PMC3110552 — target-verified WB comparison