Phospho-PKC alpha (Thr638) Recombinant Rabbit Monoclonal Antibody, Clone: [4B3], Unconjugated

Catalog Number: BYT-ORB559135
Article Name: Phospho-PKC alpha (Thr638) Recombinant Rabbit Monoclonal Antibody, Clone: [4B3], Unconjugated
Biozol Catalog Number: BYT-ORB559135
Supplier Catalog Number: orb559135
Alternative Catalog Number: BYT-ORB559135-100,BYT-ORB559135-50
Manufacturer: Biorbyt
Host: Rabbit
Category: Antikörper
Application: FC, ICC, IF, IHC-Fr, IHC-P, WB
Species Reactivity: Human, Mouse, Rat
Immunogen: KLH conjugated synthesised phosphopeptide derived from human PKC alpha around the phosphorylation site of Thr638 VL(p-T)PP
Conjugation: Unconjugated
Alternative Names: PKC alpha(T638), PKC(phospho-Thr641), phospho-PKC alpha/beta II(Thr638/641), phospho-PKC alpha/beta II(Thr638+641), PKC alpha, AAG6, Aging associated gene 6, PICK 1, PKC A, PKC alpha, PKCA, PRKACA, PRKC A, PRKCA, Protein Kinase C alpha, Protein kinase C alpha type, KPCB_HUMAN, PKC alpha.
Phospho-PKC alpha (Thr638) Recombinant Rabbit Monoclonal Antibody
Clonality: Recombinant
Concentration: 1mg/ml
Clone Designation: [4B3]
Molecular Weight: 77 kDa
UniProt: P17252
Buffer: 0.01M TBS (pH7.4) with 1% rAlbumin, 0.02% Proclin300 and 50% Glycerol.
Form: Liquid
Target: PRKCA
Application Dilute: WB=1:500-2000, IHC-P=1:50-200, IHC-F=1:50-200, ICC/IF=1:50-200, IF=1:50-200, Flow-Cyt=1:50-100
Application Notes: Modification: Phosphorylated
Blank control: MCF7. Primary Antibody (green line): Rabbit Anti-Phospho-PKC alpha (Thr638) antibody (orb559135), dilution: 2 µg/10 6 cells, Isotype Control Antibody (orange line): Rabbit IgG. Secondary Antibody: Goat anti-rabbit IgG-AF488, 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 -20C. 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 37C for 20 min, Antibody incubation with (Phospho-PKC alpha (T638) in) monoclonal Antibody, Unconjugated (orb559135) 1:50, 90 minutes at 37C, followed by a conjugated Goat Anti-Rabbit IgG antibody at 37C for 90 minutes, DAPI (blue) was used to stain the cell nuclei.
MCF-7 cell, 4% Paraformaldehyde-fixed, Triton X-100 at room temperature for 20 min, Blocking buffer (normal goat serum) at 37C for 20 min, Antibody incubation with (Phospho-PKC alpha (T638) in) monoclonal Antibody, Unconjugated (orb559135) 1:50, 90 minutes at 37C, followed by a conjugated Goat Anti-Rabbit IgG antibody at 37C for 90 minutes, DAPI (blue) was used to stain the cell nuclei.
Paraformaldehyde-fixed, paraffin embedded (human breast carcinoma), 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 37C for 30 min, Antibody incubation with (Phospho-PKC alpha (Thr638)) Monoclonal Antibody, Unconjugated (orb559135) at 1:50 overnight at 4C, followed by operating according to SP Kit (Rabbit) instructionsand 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 37C for 30 min, Antibody incubation with (Phospho-PKC alpha (Thr638)) Monoclonal Antibody, Unconjugated (orb559135) at 1:50 overnight at 4C, followed by operating according to SP Kit (Rabbit) instructionsand DAB staining.
Paraformaldehyde-fixed, paraffin embedded (rat 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 37C for 30 min, Antibody incubation with (Phospho-PKC alpha (Thr638)) Monoclonal Antibody, Unconjugated (orb559135) at 1:50 overnight at 4C, followed by operating according to SP Kit (Rabbit) instructionsand DAB staining.
Sample: Lane 1: 293 cell lysate, Lane 2: Hela cell lysate, Lane 3: Jurkat cell lysate, Primary: Anti-Phospho-PKC alpha (Thr638) (orb559135) at 1/500 dilution, Secondary: Goat Anti-Rabbit IgG - HRP at 1:5000 dilution, Predicted band size: 77 kD, Observed band size: 77 kD.
SH-SY5Y cell, 4% Paraformaldehyde-fixed, Triton X-100 at room temperature for 20 min, Blocking buffer (normal goat serum) at 37C for 20 min, Antibody incubation with (Phospho-PKC alpha (T638)) monoclonal Antibody, Unconjugated (orb559135) 1:50, 90 minutes at 37C, followed by a conjugated Goat Anti-Rabbit IgG antibody at 37C for 90 minutes, DAPI (blue) was used to stain the cell nuclei.