Phospho-IKB alpha (Tyr42) Rabbit Polyclonal Antibody

• Catalog Number: orb5520
• Category: Antibodies
• Description: Phospho-IKB alpha (Tyr42) Rabbit Polyclonal Antibody
• Species/Host: Rabbit
• Clonality: Polyclonal
• Tested applications: FC, IF, IHC-Fr, IHC-P, WB
• Predicted Reactivity: Bovine, Porcine, Rabbit, Sheep
• Reactivity: Human, Mouse
• Isotype: IgG
• Immunogen: KLH conjugated Synthesised phosphopeptide derived from human IKB alpha around the phosphorylation site of Tyr42 EE(p-Y)EQ
• Concentration: 1mg/ml
• Dilution range: WB=1:500-2000, IHC-P=1:100-500, IHC-F=1:100-500, IF=1:100-500, Flow-Cyt=1μg/Test
• Form/Appearance: Liquid
• Conjugation: Unconjugated
• MW: 35 kDa
• Target: NFKBIA
• UniProt ID: P25963
• RRID: AB_10926773
• 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: IKB alpha (phospho Y42); p-IKB alpha (phospho Y42)
• Note: For research use only
• Expiration Date: 12 months from date of receipt.

Blank control (blue line): Hela (fixed with 70% methanol (Overnight at 4°C) and then permeabilized with 90% ice-cold methanol for 20 min at -20°C). Primary Antibody (green line): Rabbit Anti-phospho-IKB alpha (Tyr42) antibody (orb5520), dilution: 1 µg/10^6 cells, Isotype Control Antibody (orange line): Rabbit IgG. Secondary Antibody (white blue line): Goat anti-rabbit IgG-PE, dilution: 1 µg/Test.

Blank control (blue line): Jurkat (fixed with 2% paraformaldehyde (10 min), then permeabilized with 90% ice-cold methanol for 30 min on ice). Primary Antibody (green line): Rabbit Anti-phospho-IKB alpha (Tyr42) antibody (orb5520), dilution: 1 µg/10^6 cells, Isotype Control Antibody (orange line): Rabbit IgG. Secondary Antibody (white blue line): Goat anti-rabbit IgG-FITC, dilution: 1 µg/Test.

Paraformaldehyde-fixed, paraffin embedded (Mouse brain), 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 (p-IKB alpha (Tyr42)) Polyclonal Antibody, Unconjugated (orb5520) at 1:400 overnight at 4°C, followed by operating according to SP Kit (Rabbit) instructionsand DAB staining.

Sample: Lane 1: HepG2 (Human) Cell Lysate at 30 ug, Lane 2: Hela (Human) Cell Lysate at 30 ug, Primary: Anti-phospho-IKB alpha (Tyr42) (orb5520) at 1/1000 dilution, Secondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilution, Predicted band size: 39 kD, Observed band size: 37 kD.

Sample: Raji (Human) Lysate at 40 ug, Primary: Anti-phospho-IKB alpha (Tyr42) (orb5520) at 1/300 dilution, Secondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilution, Predicted band size: 35 kD, Observed band size: 35 kD.

Sample: Thymus (Mouse) Lysate at 40 ug, Primary: Anti-phospho-IKB alpha (Tyr42) (orb5520) at 1/300 dilution, Secondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilution, Predicted band size: 35 kD, Observed band size: 35 kD.