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- Table of Contents
NR2F2 / COUP transcription factor 2 · Western blot design guide
Design a Western Blot for NR2F2
Real validated NR2F2 Western blot protocols, expected-band and isoform facts, troubleshooting for weak or shifted signal, and recommended anti-NR2F2 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
NR2F2 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 | ~45.6 kDa | |
| Gel | 12–15% (standard starting point) | |
| Negative control | Suggested KO / knockdown lysate |
Important caveats
Reasons your observed band may differ from the expected size.
| PTM | Phosphorylated | |
| Caveat | Phosphorylation-state controls | |
| Gene-set association | MSigDB C7 membership | |
| Isoform | 3 isoform(s) |
Section 1
Real Curated NR2F2 Western Blot Protocols
The A02420T7 protocol combines labelled catalog values with standard starting conditions. Published comparisons retain their own sample, reagent and detection scope.
| Sample / lysate | MCF-7 (40ug), HEK293T (20ug), H9C2 (20ug) (catalog A02420T7) |
| Gel % | 12–15% (standard starting point) |
| Load | 20–30 µg total protein per lane; optimize for abundance (standard starting point) |
| Transfer | Standard semi-dry transfer; verify efficiency (standard starting point) |
| Membrane | 0.45 µm PVDF (standard starting point) |
| Blocking | 5% milk or 5% BSA in TBST (standard starting point) |
| Primary antibody | A02420T7; 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 NR2F2 Western Blot Band Size?
NR2F2 is predicted at 45.6 kDa; isoforms and Thr51 phosphorylation are annotated, but their effects on band migration have not been demonstrated.
| Band near 45.6 kDa | Consistent with predicted NR2F2 mass; confirm identity with a depletion control |
| Several bands at different positions | Isoforms 1, 2 and 3 are annotated, but distinct migration is unproven |
| Single band despite three annotated isoforms | The isoforms may be unresolved or not all detected |
| Band offset from 45.6 kDa | Thr51 phosphorylation is annotated, but its migration effect is unknown |
💡Expected NR2F2 appearanceNR2F2 has a predicted mass of 45.6 kDa; no empirical band is supplied, so assess any band near that size with molecular weight markers and an NR2F2 depletion control.
| UniProt predicted mass | Sets a 45.6 kDa reference for NR2F2 |
| Splice isoform 1 | May migrate differently from other isoforms; its size is unspecified |
| Splice isoform 2 | May migrate differently from other isoforms; its size is unspecified |
| Splice isoform 3 | May migrate differently from other isoforms; its size is unspecified |
| Situation | Likely cause | Next action |
|---|---|---|
| No band in lysate | NR2F2 is nuclear and may be poorly recovered | Check nuclear enrichment and lysate recovery |
| Band higher than expected | The migration difference is unassigned by the supplied features | Check molecular weight markers and test NR2F2 depletion |
| Band lower than expected | The migration difference is unassigned; isoform sizes are unspecified | Test NR2F2 depletion and compare antibody recognition of isoforms |
| Multiple bands | Three splice isoforms are annotated, but band identities are unconfirmed | Use NR2F2 depletion to identify specific bands |
| Weak or no signal | A nuclear target may be diluted in whole-cell lysate | Check nuclear enrichment and include a positive control lysate |
Experiment evidence
Western blot images showing NR2F2 band
A02420T7 · NR2F2
A02420-1 · NR2F2
← swipe / scroll for more images →
Real WB data behind the band-size expectations in this section.
Sample controls for NR2F2 Western blot
🧪For positive controls for NR2F2 in Western blot, you can use a validated NR2F2-expressing sample once identified, since the supplied HPA record lists no positive candidate.
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: NR2F2 is nuclear, but the supplied HPA record has no tissue data to establish positive or negative controls.
HPA tissue expression evidence for NR2F2
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 NR2F2 Western Blot Tips
Deeper troubleshooting and optimisation questions for NR2F2, answered from its protein features.
How should NR2F2 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 NR2F2 isoforms produce different bands?
Isoforms · UniProt lists isoforms 1, 2 and 3. Isoform 3 lacks canonical residues 1–153; isoform 2 replaces residues 1–147 with a different sequence. Their sequence differences could affect migration, but the supplied features do not establish which bands are visible.
Check the antibody epitope against the isoform sequences. An epitope within canonical residues 1–153 is absent from isoform 3; one within residues 1–147 may also be altered in isoform 2. An antibody against an unchanged region is better suited to comparing the listed isoforms.
Check the antibody epitope against the isoform sequences. An epitope within canonical residues 1–153 is absent from isoform 3; one within residues 1–147 may also be altered in isoform 2. An antibody against an unchanged region is better suited to comparing the listed isoforms.
Can phosphorylation explain an NR2F2 band shift?
PTM · UniProt lists phosphothreonine at canonical residue 51. That residue lies in the region missing from isoform 3 and replaced in isoform 2. Use canonical UniProt numbering when comparing site claims. The listed modification alone does not establish a visible shift or explain any apparent mass difference.
Does this guide establish induction of NR2F2?
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 NR2F2?
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 A02420T7 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 should be kept consistent when quantifying NR2F2?
Quantitation · NR2F2 is listed in the nucleus, so use the same sample fraction across comparisons. Record which isoforms the antibody can detect before combining bands in one measurement. If measuring phosphorylation, distinguish a phosphothreonine 51 signal from total NR2F2 and specify that residue numbering refers to the canonical UniProt sequence.
Should NR2F2 migrate at its predicted 45.6 kDa?
Interpretation · 45.6 kDa is the predicted mass of canonical NR2F2. No observed band position is supplied, so a difference between apparent and predicted mass cannot be established. Use 45.6 kDa as a reference, not a required migration position.
First compare the antibody epitope with the three listed isoforms and their altered N-terminal sequences. NR2F2 binds DNA as a homodimer or with NR2F6 and interacts with other proteins, but those features do not establish that a higher band is a stable complex on a Western blot. Band identity needs independent validation.
First compare the antibody epitope with the three listed isoforms and their altered N-terminal sequences. NR2F2 binds DNA as a homodimer or with NR2F6 and interacts with other proteins, but those features do not establish that a higher band is a stable complex on a Western blot. Band identity needs independent validation.
Boster reagents
NR2F2 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-ARP-1 (T7) NR2F2 Antibody
Cat # A02420T7
Real WB data
Anti-ARP-1 NR2F2 Antibody
Cat # A02420-1
Two the supplier anti-NR2F2 antibodies are listed for Western blot, both with reported human, mouse, and rat reactivity and WB images. The supplied caption for A02420-1 does not identify its tested samples or antibody dilution.
Which to pick: Choose A02420T7 when its documented conditions fit your experiment: a 1:500 dilution with MCF-7, HEK293T, H9C2, and MEF whole-cell lysates. A02420-1 also has a WB image, but the supplied description gives less detail for planning.
Source: BosterBio NR2F2 gene-info card — filtered to Western-blot-capable antibodies; each card shows that product's actual WB validation figure.
References
- UniProt Consortium. UniProt entry P24468.
- Human Protein Atlas. NR2F2 tissue expression.
- PMC7336611 — target-verified WB comparison
- PMC9448590 — target-verified WB comparison
- PMC13551882 — target-verified WB comparison