RORC Rabbit Polyclonal Antibody

• Catalog Number: orb6888
• Category: Antibodies
• Description: RORC Rabbit Polyclonal Antibody
• Species/Host: Rabbit
• Clonality: Polyclonal
• Tested applications: FC, ICC, IF, IHC-Fr, IHC-P, WB
• Predicted Reactivity: Bovine, Canine, Equine, Gallus, Porcine, Rabbit
• Reactivity: Human, Mouse, Rat
• Isotype: IgG
• Immunogen: KLH conjugated synthetic peptide derived from human ROR Gamma (81-140/518aa)
• Concentration: 1mg/ml
• Dilution range: WB=1:500-2000, IHC-P=1:100-500, IHC-F=1:100-500, ICC/IF=1:100, IF=1:100-500, Flow-Cyt=1ug/test
• Form/Appearance: Liquid
• Conjugation: Unconjugated
• MW: 57 kDa
• Target: RORC
• UniProt ID: P51449
• RRID: AB_10922116
• Storage: Maintain refrigerated at 2-8°C for up to 2 weeks. For long term storage store at -20°C in small aliquots to prevent freeze-thaw cycles.
• Buffer/Preservatives: 0.01M TBS (pH7.4) with 1% rAlbumin, 0.02% Proclin300 and 50% Glycerol.
• Alternative names: RORG_HUMAN; Nuclear receptor ROR-gamma; ROR gamma;
• Note: For research use only
• Expiration Date: 12 months from date of receipt.

References

Eleftheriadis, Theodoros et al. Differential effects of the two amino acid sensing systems, the GCN2 kinase and the mTOR complex 1, on primary human alloreactive CD4⁺ T-cells Int J Mol Med, 37, 1412-1420 (2016)

PubMed: 27035541

Applications

WB

Reactivity

Human

Eleftheriadis, Theodoros et al. In Mixed Lymphocyte Reaction, the Hypoxia-Inducible Factor Prolyl-Hydroxylase Inhibitor Roxadustat Suppresses Cellular and Humoral Alloimmunity Arch Immunol Ther Exp (Warsz), 68, 31 (2020)

PubMed: 33011826

Applications

WB

Reactivity

Human

Blank control (black line): HepG2. Primary Antibody (green line): Rabbit Anti-ROR Gamma antibody (orb6888), dilution: 1 ug/Test, Secondary Antibody (white blue line): Goat anti-rabbit IgG-AF488, dilution: 0.5 ug/Test. Isotype control (orange line): Normal Rabbit IgG, Protocol, The cells were fixed with 4% PFA (10 min at room temperature) and then permeabilized with 90% ice-cold methanol for 20 min at -20°C, The cells were then incubated in 5% BSA to block non-specific protein-protein interactions for 30 min at room temperature. Cells stained with Primary Antibody for 30 min at room temperature. The secondary antibody used for 40 min at room temperature. Acquisition of 20000 events was performed.

Blank control: Molt-4. Primary Antibody (green line): Rabbit Anti-ROR Gamma antibody (orb6888), dilution: 1 µg/10^6 cells, Isotype Control Antibody (orange line): Rabbit IgG. Secondary Antibody: Goat anti-rabbit IgG-AF647, dilution: 1 µg/Test. Protocol The cells were fixed with 4% PFA (10 min at room temperature) and then permeabilized with 90% ice-cold methanol for 20 min at -20°C. The cells were then incubated in 5% BSA to block non-specific protein-protein interactions for 30 min at at room temperature. Cells stained with Primary Antibody for 30 min at room temperature. The secondary antibody used for 40 min at room temperature. Acquisition of 20000 events was performed.

HepG2 cell, 4% Paraformaldehyde-fixed, Triton X-100 at room temperature for 20 min, Blocking buffer (normal goat serum) at 37°C for 20 min, Antibody incubation with (ROR Gamma) polyclonal Antibody, Unconjugated (orb6888) 1:100, 90 minutes at 37°C, followed by a conjugated Goat Anti-Rabbit IgG antibody at 37°C for 90 minutes, DAPI (blue) was used to stain the cell nuclei.

Paraformaldehyde-fixed, paraffin embedded (mouse intestines), Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min, Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes, Blocking buffer (normal goat serum) at 37°C for 30 min, Antibody incubation with (Gamma) Polyclonal Antibody, Unconjugated (orb6888) at 1:400 overnight at 4°C, followed by a conjugated secondary for 20 minutes and DAB staining.

Sample: Lane 1: Mouse Muscle Lysates, Lane 2: Rat Muscle Lysates, Primary: Anti-ROR Gamma (orb6888) at 1/1000 dilution, Secondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilution, Predicted band size: 57 kD, Observed band size: 50 kD.

Sample: Liver (Mouse) Lysate at 40 ug, Primary: Anti-ROR Gamma (orb6888) at 1/1000 dilution, Secondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilution, Predicted band size: 57 kD, Observed band size: 50 kD.

Sample: Liver (Mouse) Lysate at 40 ug, Primary: Anti-ROR Gamma (orb6888) at 1/300 dilution, Secondary: HRP conjugated Goat-Anti-rabbit IgG (orb572747) at 1/5000 dilution, Predicted band size: 57 kD, Observed band size: 57 kD.

Tissue/Cell: mouse colon tissue, 4% Paraformaldehyde-fixed and paraffin-embedded, Antigen retrieval: citrate buffer (0.01M, pH6.0), Boiling bathing for 15 min, Block endogenous peroxidase by 3% Hydrogen peroxide for 30 min, Blocking buffer (normal goat serum) at 37°C for 20 min, Incubation: Anti-ROR Gamma Polyclonal Antibody, Unconjugated (orb6888) 1:200, overnight at 4°C, followed by conjugation to the secondary antibody and DAB staining.

Tissue/Cell: mouse colon tissue, 4% Paraformaldehyde-fixed and paraffin-embedded, Antigen retrieval: citrate buffer (0.01M, pH6.0), Boiling bathing for 15 min, Block endogenous peroxidase by 3% Hydrogen peroxide for 30 min, Blocking buffer (normal goat serum) at 37°C for 20 min, Incubation: Anti-ROR Gamma Polyclonal Antibody, Unconjugated (orb6888) 1:200, overnight at 4°C, followed by conjugation to the secondary antibody and DAB staining.