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- Table of Contents
NDP / Norrin · Western blot design guide
Design a Western Blot for NDP
Real validated NDP Western blot protocols, expected-band and isoform facts, troubleshooting for weak or shifted signal, and recommended anti-NDP 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
NDP 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 | ~15 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 | Cleaved + Disulfide-linked | |
| Caveat | Reducing-condition controls | |
| Gene-set association | MSigDB Hallmark membership | |
| Isoform | 1 isoform(s) |
Section 1
Real Curated NDP Western Blot Protocols
The A01045 protocol combines labelled catalog values with standard starting conditions. Published comparisons retain their own sample, reagent and detection scope.
| Sample / lysate | Jurkat cell lysate (catalog A01045) |
| 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 | A01045 · (A) 1 and (B) 2 μg/mL (catalog A01045) |
| 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 NDP Western Blot Band Size?
Norrin’s full-length precursor is predicted at 15 kDa; secretion, signal-peptide cleavage, and disulfide-linked dimerization could affect the blot, but no migration is empirically established.
| Band near 15 kDa | Consistent with the predicted full-length precursor; confirm identity with controls |
| Band below 15 kDa | Could reflect removal of the 1–24 signal peptide |
| Band near twice the monomer mass under nonreducing conditions | Consistent with the disulfide-linked homodimer |
| Little or no band in whole-cell lysate | Norrin is secreted |
💡Expected NDP appearanceThe full-length precursor is predicted at 15 kDa; signal-peptide cleavage may yield a smaller mature protein, and a disulfide-linked dimer may migrate higher under nonreducing conditions, but no empirical band size is supplied, so confirm identity with controls.
| Predicted full-length precursor mass | Provides a 15 kDa reference, not a measured band position |
| Signal peptide at residues 1–24 | Its cleavage yields a mature protein smaller than the precursor |
| Disulfide-linked homodimer | May yield a band near twice the monomer mass under nonreducing conditions |
| Reduction of interchain disulfides | Can shift a dimer band to monomer-sized material |
| Situation | Likely cause | Next action |
|---|---|---|
| No band in lysate | Norrin is secreted | Check conditioned medium and use an appropriate positive control |
| Band higher than expected | Disulfide-linked homodimer may persist | Compare reducing and nonreducing samples |
| Band lower than expected | Signal-peptide cleavage may remove residues 1–24 | Compare precursor and secreted samples with an identity control |
| Multiple bands | Precursor, mature protein, or incompletely reduced dimer may coexist | Compare sample fractions and reducing conditions, then verify identity |
| Weak or no signal | Secreted Norrin may be scarce in the tested fraction | Check conditioned medium and verify antibody performance with a positive control |
Experiment evidence
Western blot images showing NDP band
A01045 · NDP
← swipe / scroll for more images →
Real WB data behind the band-size expectations in this section.
Sample controls for NDP Western blot
🧪For positive controls for NDP in Western blot, you can use conditioned medium from an NDP-expressing sample; the supplied HPA data identify no positive tissue or cell line.
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 such as Ponceau alongside the samples.
⚠️Feasibility: NDP is secreted, so conditioned medium may give a stronger signal than whole-cell lysate.
HPA tissue expression evidence for NDP
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 NDP Western Blot Tips
Deeper troubleshooting and optimisation questions for NDP, answered from its protein features.
How should NDP 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.
Should multiple Norrin bands be assigned to isoforms?
Isoforms · The supplied UniProt record lists one isoform and no alternative sequence. Do not assign additional bands to annotated isoforms on this evidence alone.
Do annotated modifications prove a band shift?
PTM · The linked UniProt record describes protein features. A modification annotation alone does not demonstrate a visible shift; retain any condition or experimental qualifier attached to it.
Does this guide establish induction of NDP?
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 NDP Western blot?
Transfer · Norrin is predicted to be 15 kDa before signal-peptide cleavage. Choose and verify transfer conditions that retain low-mass proteins, checking the relevant ladder region and transferred protein on the membrane.
How should blocking be optimized?
Blocking · Standard workflow guidance: follow the A01045 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 Norrin bands be quantified across samples?
Quantitation · Keep sample reduction conditions consistent: Norrin is a disulfide-linked homodimer, so different preparation conditions may alter the band pattern. Quantify the same identified band across samples and report whether the samples came from medium or lysate.
Why might Norrin migrate below its predicted 15 kDa?
Interpretation · The 15 kDa prediction refers to the 133-residue sequence. UniProt lists a signal peptide at residues 1–24, so secreted Norrin may lack that segment. No observed band size is supplied; cleavage alone does not establish its apparent mass.
Norrin forms a disulfide-linked homodimer and has seven annotated disulfide bonds. Compare reduced and nonreduced samples to assess whether a higher band is consistent with the dimer. These features do not guarantee a particular migration pattern.
UniProt identifies Norrin as secreted. Examine conditioned medium when testing for secreted protein; a cell lysate alone may not represent the released pool.
A disulfide-linked homodimer is one possibility, especially under nonreducing conditions. Compare reduced and nonreduced samples before assigning it. The supplied record lists no glycosylation sites or modified residues, and no observed band size is available.
Norrin forms a disulfide-linked homodimer and has seven annotated disulfide bonds. Compare reduced and nonreduced samples to assess whether a higher band is consistent with the dimer. These features do not guarantee a particular migration pattern.
UniProt identifies Norrin as secreted. Examine conditioned medium when testing for secreted protein; a cell lysate alone may not represent the released pool.
A disulfide-linked homodimer is one possibility, especially under nonreducing conditions. Compare reduced and nonreduced samples before assigning it. The supplied record lists no glycosylation sites or modified residues, and no observed band size is available.
Boster reagents
NDP 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-Norrin NDP Antibody
Cat # A01045
A01045 is a human-reactive anti-NDP antibody with a Western blot image using Jurkat cell lysate at 1 and 2 μg/mL. The supplied evidence shows this sample and these conditions only; broader validation is not documented.
Which to pick: A01045 is the only listed NDP antibody. It has a Western blot image from Jurkat cell lysate, so use that sample and the reported 1–2 μg/mL concentrations as a starting point.
Source: BosterBio NDP gene-info card — filtered to Western-blot-capable antibodies; each card shows that product's actual WB validation figure.
References
- UniProt Consortium. UniProt entry Q00604.
- Human Protein Atlas. NDP tissue expression.
- PMC10981164 — target-verified WB comparison