Phospho-JNK1/2/3 (T183+T183+T221) Recombinant Rabbit Monoclonal Antibody

• Catalog Number: orb704515
• Category: Antibodies
• Description: Phospho-JNK1/2/3 (T183+T183+T221) Recombinant Rabbit Monoclonal Antibody
• Species/Host: Rabbit
• Clonality: Recombinant
• Clone Number: 6C10
• Tested applications: FC, ICC, IF, IHC-Fr, IHC-P, WB
• Predicted Reactivity: Mouse, Rat
• Reactivity: Human, Mouse, Rat
• Isotype: IgG
• Immunogen: KLH conjugated Synthesised phosphopeptide derived from human JNK1/JNK2/JNK3 around the phosphorylation site of T183/T183/T221 MM(p-T)PY
• Concentration: 1mg/ml
• Dilution range: WB=1:200-1000, IHC-P=1:100-500, IHC-F=1:100-500, ICC/IF=1:20-50, IF=1:100-500, Flow-Cyt=1:20-100
• Form/Appearance: Liquid
• Conjugation: Unconjugated
• MW: 42 kDa
• Target: MAPK8
• UniProt ID: P45983
• 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: JNK1 + JNK2 + JNK3 (phospho T183 + T183 + T221); J
• Note: For research use only
• Expiration Date: 12 months from date of receipt.

Sample: Lane 1: NIH/3T3 cell lysate, treated with Anisomycin, Lane 2: NIH/3T3 cell lysate, untreated, Primary: Anti-Phospho-JNK1/2/3 (T183+T183+T221) (orb704515) at 1:500 dilution, Secondary: Goat Anti-Rabbit IgG - HRP at 1:5000 dilution, Predicted band size: 42 kD, Observed band size: 54/46 kD.

Blank control: Hela. Primary Antibody (green line): Rabbit Anti-Phospho-JNK1/2/3 (T183+T183+T221) antibody (orb704515), dilution: 1:100, Isotype Control Antibody (orange line): Rabbit IgG. Secondary Antibody: Goat anti-rabbit IgG-AF488, dilution: 1:1000. 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-JNK1/2/3 (T183+T183+T221)) monoclonal Antibody, Unconjugated (orb704515) 1:50, 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.

HUVEC 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-JNK1/2/3 (T183+T183+T221)) monoclonal Antibody, Unconjugated (orb704515) 1:50, 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.

NIH/3T3 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-JNK1/2/3 (T183+T183+T221)) monoclonal Antibody, Unconjugated (orb704515) 1:50, 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 gastric 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 37°C for 30 min, Antibody incubation with (Phospho-JNK123 (T183+T183+T221)) Monoclonal Antibody, Unconjugated (orb704515) at 1:200 overnight at 4°C, followed by operating according to SP Kit (Rabbit) instructionsand DAB staining.

Paraformaldehyde-fixed, paraffin embedded (human prostate), 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-JNK123 (T183+T183+T221)) Monoclonal Antibody, Unconjugated (orb704515) at 1:200 overnight at 4°C, followed by operating according to SP Kit (Rabbit) instructionsand DAB staining.

Paraformaldehyde-fixed, paraffin embedded (human Uterine Corpus Cancer), 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-JNK123 (T183+T183+T221)) Monoclonal Antibody, Unconjugated (orb704515) at 1:200 overnight at 4°C, followed by operating according to SP Kit (Rabbit) instructionsand DAB staining.

Paraformaldehyde-fixed, paraffin embedded (rat kidney), 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-JNK123 (T183+T183+T221)) Monoclonal Antibody, Unconjugated (orb704515) at 1:200 overnight at 4°C, followed by operating according to SP Kit (Rabbit) instructionsand DAB staining.

Paraformaldehyde-fixed, paraffin embedded Human Heart, Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min, Antibody incubation with Phospho-JNK1 2 3 (T183+T183+T221) Monoclonal Antibody, Unconjugated (orb704515) at 1:200 overnight at 4°C, followed by conjugation to the SP Kit (Rabbit) and DAB staining.

Paraformaldehyde-fixed, paraffin embedded Human Liver, Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min, Antibody incubation with Phospho-JNK1 2 3 (T183+T183+T221) Monoclonal Antibody, Unconjugated (orb704515) at 1:200 overnight at 4°C, followed by conjugation to the SP Kit (Rabbit) and DAB staining.

Paraformaldehyde-fixed, paraffin embedded Rat Colon, Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min, Antibody incubation with Phospho-JNK1 2 3 (T183+T183+T221) Monoclonal Antibody, Unconjugated (orb704515) at 1:200 overnight at 4°C, followed by conjugation to the SP Kit (Rabbit) and DAB staining.

Paraformaldehyde-fixed, paraffin embedded Rat Heart, Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min, Antibody incubation with Phospho-JNK1 2 3 (T183+T183+T221) Monoclonal Antibody, Unconjugated (orb704515) at 1:200 overnight at 4°C, followed by conjugation to the SP Kit (Rabbit) and DAB staining.