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NR0B1 / Nuclear receptor subfamily 0 group B member 1 · Western blot design guide
Design a Western Blot for NR0B1
Source-linked NR0B1 Western blot protocol options, expected-band and isoform facts, troubleshooting for weak or shifted signal, and recommended anti-NR0B1 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
NR0B1 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 | ~51.7 kDa | |
| Observed band | ~48 kDa | |
| Gel | 5–20% (catalog A01521-2) | |
| Negative control | Suggested KO / knockdown lysate |
Important caveats
Reasons your observed band may differ from the expected size.
| PTM | — | |
| Caveat | — | |
| Gene-set association | MSigDB C7 membership | |
| Isoform | 2 isoform(s) |
Section 1
Source-Linked NR0B1 Western Blot Protocol Options
The A01521-2 protocol combines labelled catalog values with standard starting conditions. Published comparisons retain their own sample, reagent and detection scope.
| Sample / lysate | human MCF-7, human Hela, human Jurkat, human U87 (catalog A01521-2) |
| Gel % | 5–20% (catalog A01521-2) |
| Load | 30 ug; reducing conditions (catalog A01521-2) |
| Transfer | a nitrocellulose membrane at 150 mA for 50-90 minutes (catalog A01521-2) |
| Membrane | nitrocellulose membrane (catalog A01521-2) |
| Blocking | 5% non-fat milk/TBS for 1.5 hour at RT (catalog A01521-2) |
| Primary antibody | A01521-2 · 0.5 μg/mL (catalog A01521-2) |
| Primary incubation | overnight at 4°C (catalog A01521-2) |
| Secondary antibody | goat anti-rabbit IgG-HRP, 1:5000 (catalog A01521-2) |
| Secondary incubation | 1.5 hour at RT (catalog A01521-2) |
| Wash | TBS-0.1%Tween 3 times with 5 minutes each (catalog A01521-2) |
| Detection | ECL (catalog A01521-2) |
Section 2
What Is the Expected NR0B1 Western Blot Band Size?
NR0B1 is predicted at 51.7 kDa and observed near 48 kDa; the cause of the difference is not established.
| Band near 48 kDa | Empirical NR0B1 band in reducing whole-cell lysates; confirm identity with controls |
| Band near 51.7 kDa | Near the predicted protein mass; identity requires confirmation |
| Higher band near twice the monomer size | Could reflect an NR0B1 homodimer if it survives sample preparation |
| Multiple bands at different sizes | Could reflect isoforms 1 and 2, but their migration is not established |
| Weak band in one cell fraction | NR0B1 shuttles between nucleus and cytoplasm |
💡Expected NR0B1 appearanceNR0B1 has a predicted mass of 51.7 kDa, while antibody QC shows a band near 48 kDa in reducing whole-cell lysates; confirm band identity with appropriate controls.
| Predicted NR0B1 mass | 51.7 kDa by sequence, versus the empirical band near 48 kDa; the difference is unexplained |
| Splice isoform 1 | May differ in size from isoform 2; its mass and migration are unspecified |
| Splice isoform 2 | May differ in size from isoform 1; its mass and migration are unspecified |
| NR0B1 homodimer | Could appear near twice the monomer size if it remains intact during electrophoresis |
| Situation | Likely cause | Next action |
|---|---|---|
| No band in lysate | NR0B1 may be poorly recovered from the nuclear fraction | Check nuclear extraction and include a positive lysate control |
| Band higher than expected | An NR0B1 homodimer may persist during sample preparation | Compare reducing and nonreducing samples and confirm band identity |
| Band lower than expected | The observed 48 kDa band is below the 51.7 kDa prediction for an unestablished reason | Compare with a positive control and verify antibody specificity |
| Multiple bands | Isoforms 1 and 2 are documented, but distinct bands are not established | Use isoform-aware controls to assign bands |
| Weak or no signal | NR0B1 may partition between nucleus and cytoplasm | Check both fractions and include a positive lysate control |
Experiment evidence
Western blot images showing NR0B1 band
A01521-2 · NR0B1
M01521 · NR0B1
← swipe / scroll for more images →
Real WB data behind the band-size expectations in this section.
Sample controls for NR0B1 Western blot
🧪For positive controls for NR0B1 in Western blot, you can use an independently validated NR0B1-positive lysate; the supplied HPA evidence identifies 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: HPA provides no tissue candidates, so a positive lysate needs independent validation and a knockdown or KO is the practical negative control.
HPA tissue expression evidence for NR0B1
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 NR0B1 Western Blot Tips
Deeper troubleshooting and optimisation questions for NR0B1, answered from its protein features.
How should NR0B1 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.
How might NR0B1 isoforms affect band interpretation?
Isoforms · UniProt lists two isoforms. In isoform 2, residues 390–400 change from DVPGLQCVKYI to GKGKENDCNHH, and residues 401–470 are missing. These are UniProt coordinates. Isoform 2 could produce a smaller band, but these features alone do not identify the observed 48 kDa band.
Check the antibody epitope against the UniProt sequence changes. An epitope entirely within residues 401–470 of isoform 1 is absent from isoform 2; an epitope spanning residues 390–400 also differs between them.
Check the antibody epitope against the UniProt sequence changes. An epitope entirely within residues 401–470 of isoform 1 is absent from isoform 2; an epitope spanning residues 390–400 also differs between them.
Could a modification explain an unexpected NR0B1 band?
PTM · The supplied UniProt features list no modified residues or glycosylation sites. Do not assign an unexpected band to a specific modification from this record. The listed isoform sequence changes are another possibility to examine, but they do not establish band identity.
Does this guide establish induction of NR0B1?
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 NR0B1?
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 A01521-2 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.
What matters when quantifying NR0B1 across fractions?
Quantitation · NR0B1 shuttles between nucleus and cytoplasm, so specify whether measurements come from whole-cell lysate or a fraction. For comparisons across fractions, account for fraction loading and recovery before interpreting signal differences as changes in protein abundance.
Why is the observed NR0B1 band below its predicted mass?
Interpretation · The observed band is about 48 kDa, versus a predicted 51.7 kDa for the 470-residue canonical protein. The supplied features do not establish the cause of this difference; confirm band identity before assigning it to an isoform or modification.
NR0B1 is reported in both the nucleus and cytoplasm and shuttles between them. A signal in either fraction is consistent with those locations. Compare fractions when assessing localization; a change in one fraction alone need not reflect a change in total NR0B1.
UniProt reports NR0B1 homodimers and interactions with other proteins. Those interactions make a complex worth considering, but they do not establish that a higher Western blot band contains one. Verify its identity before assigning it to a dimer or binding partner.
NR0B1 is reported in both the nucleus and cytoplasm and shuttles between them. A signal in either fraction is consistent with those locations. Compare fractions when assessing localization; a change in one fraction alone need not reflect a change in total NR0B1.
UniProt reports NR0B1 homodimers and interactions with other proteins. Those interactions make a complex worth considering, but they do not establish that a higher Western blot band contains one. Verify its identity before assigning it to a dimer or binding partner.
Boster reagents
NR0B1 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-DAX-1/NR0B1 Antibody Picoband®
Cat # A01521-2
Real WB data
Anti-NR0B1 / DAX1 Monoclonal Antibody
Cat # M01521
Both listed anti-NR0B1 antibodies have Western blot images from human cell lysates, with reported bands near the expected 48 kDa. The supplied evidence covers the named lysates and product images; no independent publication evidence is provided.
Which to pick: Choose A01521-2 for its reported WB results in MCF-7, HeLa, Jurkat, or U87 lysates, or M01521 for A549 or RT4 lysates. Both list human reactivity and have WB images; select based on your sample context and preferred polyclonal or monoclonal format.
Source: BosterBio NR0B1 gene-info card — filtered to Western-blot-capable antibodies; each card shows that product's actual WB validation figure.
References
- UniProt Consortium. UniProt entry P51843.
- Human Protein Atlas. NR0B1 tissue expression.
- PMC8345183 — target-verified WB comparison
- PMC4525203 — target-verified WB comparison
- PMC12391575 — target-verified WB comparison