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- Table of Contents
NRP2 / Neuropilin-2 · Western blot design guide
Design a Western Blot for NRP2
Real validated NRP2 Western blot protocols, expected-band and isoform facts, troubleshooting for weak or shifted signal, and recommended anti-NRP2 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
NRP2 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 | ~104.8 kDa | |
| Observed band | 105 kDa | |
| Gel | 8–10% (standard starting point) | |
| Positive control | Adipose tissue (IHC candidate; verify WB) +4 more | |
| Negative control | Appendix (IHC candidate; verify WB) |
Important caveats
Reasons your observed band may differ from the expected size.
| PTM | Glycosylated + Cleaved | |
| Caveat | Glycosylation-state controls | |
| Gene-set association | MSigDB Hallmark membership | |
| Isoform | 6 isoform(s) |
Section 1
Real Curated NRP2 Western Blot Protocols
The PA1771 protocol combines labelled catalog values with standard starting conditions. Published comparisons retain their own sample, reagent and detection scope.
| Sample / lysate | Rat Brain at 50ug, Rat Testis at 50ug Predicted bind size: 105KD Observed bind size: 105KD (catalog PA1771) |
| Gel % | 8–10% (standard starting point) |
| Load | 20–30 µg total protein per lane; optimize for abundance (standard starting point) |
| Transfer | Wet/tank transfer; optimize duration (standard starting point) |
| Membrane | 0.45 µm PVDF (standard starting point) |
| Blocking | 5% milk or 5% BSA in TBST (standard starting point) |
| Primary antibody | PA1771; 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 NRP2 Western Blot Band Size?
NRP2 is predicted at 104.8 kDa and observed at 105 kDa; the cause of this small difference is not established.
| Band near 105 kDa | Matches the empirical NRP2 band; confirm identity with antibody controls. |
| Band slightly below the full-length precursor | Could reflect cleavage of the 1–20 signal peptide; its migration is unmeasured. |
| Band above 105 kDa | Could reflect N-linked glycosylation, but the sites do not establish a visible shift. |
| Several bands at different positions | Could include NRP2 isoforms; distinct migration has not been demonstrated. |
💡Expected NRP2 appearanceNRP2 has a predicted mass of 104.8 kDa and an empirical band at 105 kDa; confirm band identity with ordinary antibody controls because the supplied features do not establish its migration mechanism.
| Predicted 104.8-kDa mass | Provides the sequence-based reference for the observed 105-kDa band. |
| N-linked glycosylation at Asn152 and Asn157 | May affect apparent size; a visible shift is not established. |
| N-linked glycosylation at Asn629 and Asn839 | May affect apparent size; a visible shift is not established. |
| Signal peptide at residues 1–20 | Cleavage could make the mature chain smaller than its precursor; migration is unmeasured. |
| Isoforms A22, A0, A17, B0, B5, and s9 | May differ in size, but distinct band positions are not established. |
| Situation | Likely cause | Next action |
|---|---|---|
| No band in lysate | Membrane NRP2 may be poorly extracted. | Check membrane extraction and a positive tissue lysate. |
| Band higher than expected | N-linked glycans may affect migration; the shift is unverified. | Compare treated and untreated samples and confirm band identity. |
| Band lower than expected | Signal-peptide cleavage could reduce size, but its band position is unknown. | Confirm identity with a second antibody and a positive control. |
| Broad smear instead of sharp band | Variable N-linked glycosylation is possible but unproven here. | Compare glycosidase-treated and untreated samples. |
| Multiple bands | NRP2 has six named isoforms, though separate bands are unverified. | Check isoform expression and antibody specificity. |
| Weak or no signal | Membrane extraction or low NRP2 abundance may limit detection. | Check extraction, loading, and a positive tissue lysate. |
Experiment evidence
Western blot images showing NRP2 band
PA1771 · NRP2
← swipe / scroll for more images →
Real WB data behind the band-size expectations in this section.
Sample controls for NRP2 Western blot
🧪HPA-IHC candidate guidance (verify in WB): For positive controls for NRP2 in Western blot, you can use adipose tissue, which HPA scores High.
Positive control: Adipose tissue (IHC candidate; verify WB)
Negative control: Appendix (IHC candidate; verify WB)
Loading controls: Run GAPDH, β-actin, and a total-protein stain alongside the samples.
⚠️Feasibility: Because NRP2 is membrane-associated, membrane-enriched lysate may improve detection.
HPA tissue expression evidence for NRP2
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 |
|---|---|---|---|---|
| Adipose tissue | adipocytes | High | Protein (IHC) | HPA → |
| Breast | adipocytes | High | Protein (IHC) | HPA → |
| Cerebral cortex | endothelial cells | High | Protein (IHC) | HPA → |
| Placenta | trophoblastic cells | High | Protein (IHC) | HPA → |
| Skin | keratinocytes | High | Protein (IHC) | HPA → |
Lower expression tissues · IHC evidence, not confirmed WB-negative controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| Appendix | glandular cells | Not detected | Protein (IHC) | HPA → |
| Bone marrow | hematopoietic cells | Not detected | Protein (IHC) | HPA → |
| Endometrium | cells in endometrial stroma | Not detected | Protein (IHC) | HPA → |
| Fallopian tube | glandular cells | Not detected | Protein (IHC) | HPA → |
| Liver | cholangiocytes | Not detected | Protein (IHC) | HPA → |
Section 3
Advanced NRP2 Western Blot Tips
Deeper troubleshooting and optimisation questions for NRP2, answered from its protein features.
How should NRP2 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 NRP2 isoforms produce different Western blot bands?
Isoforms · Six isoforms are listed: A22, A0, A17, B0, B5 and s9. Their sequence differences could affect band size or antibody recognition. In particular, s9 lacks canonical residues 556–931; a C-terminal antibody directed within that region would not recognize that sequence in s9.
Which NRP2 glycosylation sites matter when interpreting band shifts?
PTM · UniProt lists N-linked sites at Asn152, Asn157, Asn629 and Asn839, using canonical sequence coordinates. Glycosylation could affect migration, but the listed sites do not establish that a particular band has shifted.
Does this guide establish induction of NRP2?
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 NRP2?
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 PA1771 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 NRP2 bands be quantified across samples?
Quantitation · Use a consistently defined band and an antibody epitope present in the isoforms you intend to compare. Because NRP2 has six annotated isoforms and four N-linked glycosylation sites, document whether quantification includes one band or several; band intensity alone does not identify an isoform.
Why is the observed NRP2 band near its predicted mass?
Interpretation · The reported band is 105 kDa, close to the 104.8 kDa predicted mass. NRP2 has a signal peptide at residues 1–20 and four annotated N-linked glycosylation sites, but these features alone cannot explain the small difference or establish a visible shift.
Check the antibody epitope against the isoform being measured. A0 lacks canonical residues 809–830; A17 and B0 lack 809–813. B0 and B5 replace the canonical sequence at 814–931, and s9 replaces 548–555 and lacks 556–931. These are UniProt canonical coordinates, which may differ from antibody numbering.
First check whether the antibody epitope is retained in the relevant isoform. The listed sequence changes, especially s9's loss of residues 556–931, offer specific possibilities. N-linked glycosylation and six annotated disulfide bonds may also affect migration, but these features alone cannot assign an unexpected band.
NRP2 has a signal peptide at canonical residues 1–20. Keep that segment in mind when comparing the 104.8 kDa predicted mass with a mature protein band. The supplied features do not establish its precise effect on the observed 105 kDa band.
Check the antibody epitope against the isoform being measured. A0 lacks canonical residues 809–830; A17 and B0 lack 809–813. B0 and B5 replace the canonical sequence at 814–931, and s9 replaces 548–555 and lacks 556–931. These are UniProt canonical coordinates, which may differ from antibody numbering.
First check whether the antibody epitope is retained in the relevant isoform. The listed sequence changes, especially s9's loss of residues 556–931, offer specific possibilities. N-linked glycosylation and six annotated disulfide bonds may also affect migration, but these features alone cannot assign an unexpected band.
NRP2 has a signal peptide at canonical residues 1–20. Keep that segment in mind when comparing the 104.8 kDa predicted mass with a mature protein band. The supplied features do not establish its precise effect on the observed 105 kDa band.
Boster reagents
NRP2 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-Neuropilin-2 NRP2 Antibody Picoband®
Cat # PA1771
PA1771 is the sole listed anti-NRP2 antibody, with reported Human, Mouse, and Rat reactivity. Its WB image shows rat brain and testis lysates (50 µg per lane) at 0.5 µg/mL antibody, with a 105 kDa band. No Human or Mouse WB image is supplied.
Which to pick: Only PA1771 is listed. Its WB image supports choosing it for rat brain or testis lysates under the reported conditions. For Human or Mouse samples, reactivity is listed, but no corresponding WB image is provided.
Source: BosterBio NRP2 gene-info card — filtered to Western-blot-capable antibodies; each card shows that product's actual WB validation figure.
References
- UniProt Consortium. UniProt entry O60462.
- Human Protein Atlas. NRP2 tissue expression.
- PMC7509939 — target-verified WB comparison
- PMC12340832 — target-verified WB comparison
- PMC8917722 — target-verified WB comparison