Phospho-IKK alpha (Thr23) Rabbit Polyclonal Antibody

• Catalog Number: orb5512
• Category: Antibodies
• Description: Phospho-IKK alpha (Thr23) Rabbit Polyclonal Antibody
• Species/Host: Rabbit
• Clonality: Polyclonal
• Tested applications: FC, ICC, IF, IHC-Fr, IHC-P
• Predicted Reactivity: Bovine, Canine, Gallus, Porcine, Rabbit, Rat
• Reactivity: Human, Mouse
• Isotype: IgG
• Immunogen: KLH conjugated Synthesised phosphopeptide derived from human IKK alpha around the phosphorylation site of Thr23 LG(p-T)GG
• Concentration: 1mg/ml
• Dilution range: IHC-P=1:100-500, IHC-F=1:100-500, ICC/IF=1:100, IF=1:100-500, Flow-Cyt=2ug/Test
• Form/Appearance: Liquid
• Conjugation: Unconjugated
• MW: 85 kDa
• Target: CHUK
• UniProt ID: O15111
• RRID: AB_10924144
• 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: IKK alpha (phospho T23); p-IKK alpha (phospho T23)
• Note: For research use only
• Expiration Date: 12 months from date of receipt.

Blank control (black line): U251. Primary Antibody (green line): Rabbit Anti-Phospho-IKK alpha (Thr23) antibody (orb5512), dilution: 2 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.

Hela 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 (Phospho-IKK alpha (Thr23)) polyclonal Antibody, Unconjugated (orb5512) 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 (Human glioma), 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 (Phospho-IKK alpha (Thr23)) Polyclonal Antibody, Unconjugated (orb5512) at 1:400 overnight at 4°C, followed by a conjugated secondary antibody for 20 minutes and DAB staining.

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 (Phospho-IKK alpha (Thr23)) Polyclonal Antibody, Unconjugated (orb5512) at 1:400 overnight at 4°C, followed by a conjugated secondary antibody for 20 minutes and DAB staining.