Phospho-IKK beta (Ser466) Rabbit Polyclonal Antibody

• Catalog Number: orb5518
• Category: Antibodies
• Description: Phospho-IKK beta (Ser466) Rabbit Polyclonal Antibody
• Species/Host: Rabbit
• Clonality: Polyclonal
• Tested applications: FC, ICC, IF, IHC-Fr, IHC-P
• Predicted Reactivity: Bovine, Canine, Porcine, Rabbit
• Reactivity: Human, Mouse, Rat
• Isotype: IgG
• Immunogen: KLH conjugated Synthesised phosphopeptide derived from human IKK beta around the phosphorylation site of Ser466 CL(p-S)KM
• Antibody Type: Primary Antibody
• 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=1ug/Test
• Form/Appearance: Liquid
• Conjugation: Unconjugated
• MW: 87 kDa
• Target: IKBKB
• UniProt ID: O14920
• RRID: AB_10935693
• 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 beta(phospho S466); IKK beta(phospho Ser466);
• Note: For research use only
• Expiration Date: 12 months from date of receipt.

A549 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 beta (Ser466)) polyclonal Antibody, Unconjugated (orb5518) 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.

Blank control: A549. Primary Antibody (green line): Rabbit Anti-phospho-IKK beta (Ser466) antibody (orb5518), 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 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.

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 beta (Ser466)) Polyclonal Antibody, Unconjugated (orb5518) at 1:500 overnight at 4°C, followed by a conjugated secondary for 20 minutes and DAB staining.

Tissue/Cell: rat brain tissue, 4% Paraformaldehyde-fixed and paraffin-embedded, Antigen retrieval: citrate buffer (0.01M, pH 6.0), 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-phospho-IKK beta (Ser466) Polyclonal Antibody, Unconjugated (orb5518) 1:200, overnight at 4°C, followed by conjugation to the secondary antibody and DAB staining.