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- Table of Contents
PLP2 / Proteolipid protein 2 · Western blot design guide
Design a Western Blot for PLP2
Real validated PLP2 Western blot protocols, expected-band and isoform facts, troubleshooting for weak or shifted signal, and recommended anti-PLP2 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
PLP2 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 | ~16.7 kDa | |
| Observed band | ~17 kDa | |
| Gel | 5–20% (catalog A06255-1) | |
| Positive control | Cerebellum (IHC candidate; verify WB) +4 more | |
| Negative control | Ovary (IHC candidate; verify WB) |
Important caveats
Reasons your observed band may differ from the expected size.
| PTM | Glycosylated | |
| Caveat | Glycosylation-state controls | |
| Gene-set association | MSigDB C7 membership | |
| Isoform | 2 isoform(s) |
Section 1
Real Curated PLP2 Western Blot Protocols
The A06255-1 protocol combines labelled catalog values with standard starting conditions. Published comparisons retain their own sample, reagent and detection scope.
| Sample / lysate | human COLO320, human A431 (catalog A06255-1) |
| Gel % | 5–20% (catalog A06255-1) |
| Load | 30 ug; reducing conditions (catalog A06255-1) |
| Transfer | a nitrocellulose membrane at 150 mA for 50-90 minutes (catalog A06255-1) |
| Membrane | nitrocellulose membrane (catalog A06255-1) |
| Blocking | 5% non-fat milk/TBS for 1.5 hour at RT (catalog A06255-1) |
| Primary antibody | A06255-1 · 0.5 μg/mL (catalog A06255-1) |
| Primary incubation | overnight at 4°C (catalog A06255-1) |
| Secondary antibody | goat anti-rabbit IgG-HRP, 1:5000 (catalog A06255-1) |
| Secondary incubation | 1.5 hour at RT (catalog A06255-1) |
| Wash | TBS-0.1%Tween 3 times with 5 minutes each (catalog A06255-1) |
| Detection | ECL (catalog A06255-1) |
Section 2
What Is the Expected PLP2 Western Blot Band Size?
PLP2 is predicted at 16.7 kDa and observed at ~17 kDa; the cause of their small difference is not established.
| Band at ~17 kDa | Empirical PLP2 band in reducing whole-cell lysates, close to the 16.7 kDa predicted mass |
| Band above ~17 kDa | N-linked glycosylation at Asn18 or Asn108 could affect migration; a visible shift is unproven |
| Several bands | PLP2 has isoforms 1 and 2, but distinct band positions are unestablished |
| Faint or absent band in whole-cell lysate | PLP2 is a multi-pass membrane protein that may be poorly recovered |
💡Expected PLP2 appearancePLP2 has a predicted mass of 16.7 kDa and an empirical band at ~17 kDa in reducing whole-cell lysates; confirm band identity with antibody specificity controls.
| UniProt predicted mass | 16.7 kDa predicted; a PLP2 band was observed at ~17 kDa |
| N-linked glycosylation at Asn18 | Could alter apparent size; no site-specific shift is established |
| N-linked glycosylation at Asn108 | Could alter apparent size; no site-specific shift is established |
| Isoforms 1 and 2 | May differ in size, but their relative masses and band separation are unknown |
| Situation | Likely cause | Next action |
|---|---|---|
| No band in lysate | Multi-pass membrane PLP2 may be poorly extracted | Check membrane solubilization and test a membrane-enriched fraction |
| Band higher than expected | N-linked glycosylation could affect migration, but its effect here is unproven | Compare treated and untreated samples after N-glycan removal and verify band identity |
| Band lower than expected | Isoform identity or band specificity is uncertain | Use an isoform-aware reference and antibody specificity control |
| Broad smear instead of sharp band | Variation in N-linked glycans is possible but unestablished | Compare samples before and after N-glycan removal |
| Multiple bands | Isoforms 1 and 2 are annotated, but distinct migration is unproven | Check antibody specificity and compare isoform-aware references |
| Weak or no signal | Membrane PLP2 may be poorly recovered | Check extraction and sample loading with a membrane protein control |
Experiment evidence
Western blot images showing PLP2 band
A06255-1 · PLP2
← swipe / scroll for more images →
Real WB data behind the band-size expectations in this section.
Sample controls for PLP2 Western blot
🧪HPA-IHC candidate guidance (verify in WB): For positive controls for PLP2 in Western blot, you can use cerebellum lysate, which has high HPA expression.
Positive control: Cerebellum (IHC candidate; verify WB)
Negative control: Ovary (IHC candidate; verify WB)
Loading controls: Run GAPDH, β-actin, and a total-protein stain alongside the samples.
⚠️Feasibility: As PLP2 is a multi-pass membrane protein, membrane-enriched lysate may improve detection.
HPA tissue expression evidence for PLP2
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 |
|---|---|---|---|---|
| Cerebellum | Purkinje cells | High | Protein (IHC) | HPA → |
| Duodenum | glandular cells | High | Protein (IHC) | HPA → |
| Gallbladder | glandular cells | High | Protein (IHC) | HPA → |
| Kidney | cells in tubules | High | Protein (IHC) | HPA → |
| Liver | hepatocytes | High | Protein (IHC) | HPA → |
Lower expression tissues · IHC evidence, not confirmed WB-negative controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| Ovary | ovarian stroma cells | Not detected | Protein (IHC) | HPA → |
| Bronchus | respiratory epithelial cells | Low | Protein (IHC) | HPA → |
| Endometrium | glandular cells | Low | Protein (IHC) | HPA → |
| Epididymis | glandular cells | Low | Protein (IHC) | HPA → |
| Esophagus | squamous epithelial cells | Low | Protein (IHC) | HPA → |
Section 3
Advanced PLP2 Western Blot Tips
Deeper troubleshooting and optimisation questions for PLP2, answered from its protein features.
How should PLP2 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.
Can PLP2 isoforms produce different bands?
Isoforms · PLP2 has isoforms 1 and 2. In isoform 2, the canonical sequence at residues 117–152 is replaced by a different sequence. This may alter migration, but the features do not establish distinct band positions.
The isoforms differ in the region corresponding to canonical residues 117–152. An antibody recognizing that region may detect the isoforms differently. Check the antibody's stated epitope before interpreting a missing isoform band; the coordinates here refer to the UniProt canonical sequence.
The isoforms differ in the region corresponding to canonical residues 117–152. An antibody recognizing that region may detect the isoforms differently. Check the antibody's stated epitope before interpreting a missing isoform band; the coordinates here refer to the UniProt canonical sequence.
Could glycosylation affect PLP2 migration?
PTM · UniProt lists N-linked glycosylation at Asn18 and Asn108, using canonical sequence coordinates. These sites could affect migration, but their annotation alone does not establish a visible shift.
Does this guide establish induction of PLP2?
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 PLP2 Western blot?
Transfer · PLP2 is a small, multi-pass membrane protein with a predicted mass of 16.7 kDa. Optimize transfer for retention of a protein near 17 kDa and verify recovery on the membrane. The supplied features do not specify one transfer method.
How should blocking be optimized?
Blocking · Standard workflow guidance: follow the A06255-1 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.
How should PLP2 bands be quantified?
Quantitation · PLP2 has two isoforms and two annotated glycosylation sites. Define which band or bands are being measured, then apply that choice consistently across samples. The annotations alone do not identify which molecular form produces each band.
How should the ~17 kDa band compare with predicted PLP2 mass?
Interpretation · The observed ~17 kDa band is close to the 16.7 kDa predicted mass. The listed features do not establish a cause for any small difference in migration.
Consider the isoform 2 sequence change at canonical residues 117–152 and N-linked glycosylation at Asn18 and Asn108. Neither annotation establishes the identity of an unexpected band or proves that a visible shift should occur.
Consider the isoform 2 sequence change at canonical residues 117–152 and N-linked glycosylation at Asn18 and Asn108. Neither annotation establishes the identity of an unexpected band or proves that a visible shift should occur.
Boster reagents
PLP2 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-PLP2 Antibody Picoband®
Cat # A06255-1
The catalog reports one anti-PLP2 Western blot antibody, A06255-1, with stated human reactivity. Its WB image reports a band near the expected 17 kDa in human COLO320 and A431 whole-cell lysates under reducing conditions. Evidence is limited to these reported samples and conditions.
Which to pick: A06255-1 is the only listed option and has a WB image. For similar human lysates, its reported 0.5 μg/mL primary antibody concentration is a starting point; performance in other samples is not shown.
Source: BosterBio PLP2 gene-info card — filtered to Western-blot-capable antibodies; each card shows that product's actual WB validation figure.
References
- UniProt Consortium. UniProt entry Q04941.
- Human Protein Atlas. PLP2 tissue expression.
- PMC7077595 — target-verified WB comparison
- PMC10548289 — target-verified WB comparison
- PMC4616445 — target-verified WB comparison