Phospho-BTK (Tyr223) Rabbit Polyclonal Antibody

• Catalog Number: orb5651
• Category: Antibodies
• Description: Phospho-BTK (Tyr223) Rabbit Polyclonal Antibody
• Species/Host: Rabbit
• Clonality: Polyclonal
• Tested applications: ELISA, FC, IF, IHC-Fr, IHC-P
• Predicted Reactivity: Bovine, Equine, Porcine, Rat
• Reactivity: Human, Mouse, Rabbit
• Isotype: IgG
• Immunogen: KLH conjugated Synthesised phosphopeptide derived from human Btk around the phosphorylation site of Tyr223 AL(p-Y)DY
• Antibody Type: Primary Antibody
• Concentration: 1mg/ml
• Dilution range: IHC-P=1:100-500, IHC-F=1:100-500, IF=1:100-500, Flow-Cyt=1ug/test, ELISA=1:5000-10000
• Form/Appearance: Liquid
• Conjugation: Unconjugated
• MW: 76 kDa
• Target: BTK
• UniProt ID: Q06187
• RRID: AB_10932256
• 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: BTK(phospho Y223); p-BTK(phospho Y223); Agammaglob
• Note: For research use only
• Expiration Date: 12 months from date of receipt.

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-BTK (Tyr223)) Polyclonal Antibody, Unconjugated (orb5651) at 1:400 overnight at 4°C, followed by operating according to SP Kit (Rabbit) instructions and DAB staining.

Paraformaldehyde-fixed, paraffin embedded (Mouse colon), 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-BTK (Tyr223)) Polyclonal Antibody, Unconjugated (orb5651) at 1:400 overnight at 4°C, followed by operating according to SP Kit (Rabbit) instructions and DAB staining.

Paraformaldehyde-fixed, paraffin embedded (Mouse lung), 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-BTK (Tyr223)) Polyclonal Antibody, Unconjugated (orb5651) at 1:400 overnight at 4°C, followed by operating according to SP Kit (Rabbit) instructions and DAB staining.

phosphopeptide, non phosphopeptide.

Tissue/Cell: human lung carcinoma, 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-Phospho-BTK (Tyr223) Polyclonal Antibody, Unconjugated (orb5651) 1:200, overnight at 4°C, followed by conjugation to the secondary antibody and DAB staining.

Tissue/Cell: mouse spleen 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-Phospho-BTK (Tyr223) Polyclonal Antibody, Unconjugated (orb5651) 1:200, overnight at 4°C, followed by conjugation to the secondary antibody and DAB staining.

U-937 cells were fixed with 4% PFA for 10 min at room temperature, permeabilized with 90% ice-cold methanol for 20 min at room temperature, and incubated in 5% BSA blocking buffer for 30 min at room temperature. Cells were then stained with Phospho-BTK (Tyr223) Antibody (orb5651) at 1:100 dilution in blocking buffer and incubated for 30 min at room temperature, washed twice with 2% BSA in PBS, followed by secondary antibody incubation for 40 min at room temperature. Acquisitions of 20000 events were performed. Cells stained with primary antibody (green), and isotype control (orange).