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- Table of Contents
RORC / Nuclear receptor ROR-gamma · Western blot design guide
Design a Western Blot for RORC
Real validated RORC Western blot protocols, expected-band and isoform facts, troubleshooting for weak or shifted signal, and recommended anti-RORC 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
RORC 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 | ~58.2 kDa | |
| Observed band | ~58 kDa | |
| Gel | 5–20% (catalog A00444-3) | |
| 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
Real Curated RORC Western Blot Protocols
The A00444-3 protocol combines labelled catalog values with standard starting conditions. Published comparisons retain their own sample, reagent and detection scope.
| Sample / lysate | human HCCT, human MOLT-4 (catalog A00444-3) |
| Gel % | 5–20% (catalog A00444-3) |
| Load | 30 ug; reducing conditions (catalog A00444-3) |
| Transfer | a nitrocellulose membrane at 150 mA for 50-90 minutes (catalog A00444-3) |
| Membrane | nitrocellulose membrane (catalog A00444-3) |
| Blocking | 5% non-fat milk/TBS for 1.5 hour at RT (catalog A00444-3) |
| Primary antibody | A00444-3 · 0.5 μg/mL (catalog A00444-3) |
| Primary incubation | overnight at 4°C (catalog A00444-3) |
| Secondary antibody | goat anti-rabbit IgG-HRP, 1:5000 (catalog A00444-3) |
| Secondary incubation | 1.5 hour at RT (catalog A00444-3) |
| Wash | TBS-0.1%Tween 3 times with 5 minutes each (catalog A00444-3) |
| Detection | ECL (catalog A00444-3) |
Section 2
What Is the Expected RORC Western Blot Band Size?
RORC is predicted at 58.2 kDa and observed near 58 kDa; the small difference has no established cause, and isoform migration is unknown.
| Band near 58 kDa | consistent with the observed RORC band and the 58.2 kDa prediction |
| Band at another position | could reflect isoform 1 or 2, but its identity needs confirmation |
| Two bands at different positions | could represent both named isoforms if each band is confirmed |
| Little or no band in a cytoplasmic fraction | consistent with RORC's nuclear location |
💡Expected RORC appearanceRORC is predicted at 58.2 kDa and has an empirical band near 58 kDa; confirm band identity with antibody and sample controls because isoform migration is not established.
| UniProt predicted mass | places the reference protein near 58.2 kDa |
| 518-residue reference sequence | defines the sequence underlying the predicted 58.2 kDa mass |
| Isoform 1 | may have a different apparent size from isoform 2; its mass is not supplied |
| Isoform 2 | may have a different apparent size from isoform 1; its mass is not supplied |
| Situation | Likely cause | Next action |
|---|---|---|
| No band in lysate | nuclear RORC may be poorly recovered | check nuclear extraction and a positive control |
| Band higher than expected | band identity is uncertain; no size-raising modification is listed | compare a positive control and verify antibody specificity |
| Band lower than expected | an isoform or an unrelated band may be detected | compare isoform expression and verify band identity |
| Multiple bands | isoforms 1 and 2 are listed, but their migration is unknown | check band identity with isoform-aware controls |
| Weak or no signal | RORC abundance or nuclear recovery may be low | check a positive control and nuclear extract |
| Fragments below expected size | sample degradation is possible; no cleavage feature is listed | prepare fresh lysate and compare with a positive control |
Experiment evidence
Western blot images showing RORC band
A00444-3 · RORC
← swipe / scroll for more images →
Real WB data behind the band-size expectations in this section.
Sample controls for RORC Western blot
🧪For positive controls for RORC in Western blot, you can use no HPA-supported tissue or cell sample because no HPA data were supplied.
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: HPA data are unavailable, so tissue controls cannot be selected; RORC is nuclear.
HPA tissue expression evidence for RORC
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 RORC Western Blot Tips
Deeper troubleshooting and optimisation questions for RORC, answered from its protein features.
How should RORC 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 RORC isoforms produce different bands?
Isoforms · UniProt lists two isoforms. Relative to isoform 1, isoform 2 lacks residues 1–21 and changes residues 22–24 from HTS to MRT. This sequence difference could affect migration, but the features do not establish a visible separation.
Check the antibody epitope against the isoform sequences. An epitope within isoform 1 residues 1–21 is absent from isoform 2; residues 22–24 also differ. UniProt coordinates here refer to isoform 1.
Check the antibody epitope against the isoform sequences. An epitope within isoform 1 residues 1–21 is absent from isoform 2; residues 22–24 also differ. UniProt coordinates here refer to isoform 1.
Do the supplied modifications explain a shifted RORC band?
PTM · No modified residues or glycosylation sites are listed in the supplied features. These data do not establish a modification-driven shift or explain a difference between apparent and predicted mass.
Does this guide establish induction of RORC?
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 RORC?
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 A00444-3 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 RORC bands be quantified across samples?
Quantitation · Use the same sample fraction and band selection across samples. Because RORC has two annotated isoforms, define whether quantitation includes one resolved band or both before comparing signals.
Does the ~58 kDa band match predicted RORC mass?
Interpretation · The observed ~58 kDa band is close to the supplied 58.2 kDa predicted mass. That agreement supports the assignment but does not establish band identity on its own.
RORC is annotated as nuclear. A nuclear fraction can help assess a suspected RORC band; account for fraction choice when comparing samples.
Compare its position with the ~58 kDa observed band and consider the two annotated isoforms. Their sequence differences make an additional band possible but do not identify it; band position alone cannot establish its identity.
RORC is annotated as nuclear. A nuclear fraction can help assess a suspected RORC band; account for fraction choice when comparing samples.
Compare its position with the ~58 kDa observed band and consider the two annotated isoforms. Their sequence differences make an additional band possible but do not identify it; band position alone cannot establish its identity.
Boster reagents
RORC 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-ROR gamma/RORC Antibody Picoband®
Cat # A00444-3
The catalog reports one anti-RORC antibody, A00444-3, with a Western blot image showing an approximately 58 kDa band in human HCCT tissue and MOLT-4 cell lysates. The supplied evidence is limited to this reported blot.
Which to pick: A00444-3 is the only listed option. Its stated reactivity is human, and its WB image uses human HCCT tissue and MOLT-4 cell lysates at 0.5 μg/mL primary antibody.
Source: BosterBio RORC gene-info card — filtered to Western-blot-capable antibodies; each card shows that product's actual WB validation figure.
References
- UniProt Consortium. UniProt entry P51449.
- Human Protein Atlas. RORC tissue expression.
- PMC12837284 — target-verified WB comparison