ERK1/2 Recombinant Rabbit Monoclonal Antibody

• Catalog Number: orb704524
• Category: Antibodies
• Description: ERK1/2 Recombinant Rabbit Monoclonal Antibody
• Species/Host: Rabbit
• Clonality: Recombinant
• Clone Number: 3A12
• Tested applications: FC, ICC, IF, IHC-Fr, IHC-P, WB
• Predicted Reactivity: Zebrafish
• Reactivity: Human, Mouse, Rat
• Isotype: IgG
• Immunogen: KLH conjugated synthetic peptide derived from human ERK1/2
• Concentration: 1mg/ml
• Dilution range: WB=1:500-2000, 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: 42/44 kDa
• Target: MAPK3
• UniProt ID: P28482
• 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: ERK1 + ERK2; ERK 1; ERK 2; ERK-2; ERK1; ERK2; ERT1
• Note: For research use only
• Expiration Date: 12 months from date of receipt.

25 ug total protein per lane of various lysates probed with ERK1/2 monoclonal antibody, unconjugated (orb704524) at 1:1000 dilution and 4°C overnight incubation. Followed by conjugated secondary antibody incubation at r.t. for 60 min.

Blank control: Hela. Primary Antibody (green line): Rabbit Anti-ERK1/2 antibody (orb704524), dilution: 1 µg/10^6 cells, Isotype Control Antibody (orange line): Rabbit IgG. Secondary Antibody: Goat anti-rabbit IgG-AF647, 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 -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.

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 (ERK1 2) Monoclonal Antibody, Unconjugated (orb704524) at 1:200 overnight at 4°C, 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 37°C for 30 min, Antibody incubation with (ERK1 2) Monoclonal Antibody, Unconjugated (orb704524) at 1:200 overnight at 4°C, followed by operating according to SP Kit (Rabbit) instructionsand DAB staining.

Paraformaldehyde-fixed, paraffin embedded (rat 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 (ERK1 2) Monoclonal Antibody, Unconjugated (orb704524) at 1:200 overnight at 4°C, followed by operating according to SP Kit (Rabbit) instructionsand DAB staining.

Sample: Lane 1: Cerebrum (Mouse) Lysate at 40 ug, Lane 2: Pancreas (Mouse) Lysate at 40 ug, Lane 3: Large intestine (Mouse) Lysate at 40 ug, Lane 4: Lymph node (Mouse) Lysate at 40 ug, Lane 5: Cerebrum (Rat) Lysate at 40 ug, Lane 6: Pancreas (Rat) Lysate at 40 ug, Lane 7: Large intestine (Rat) Lysate at 40 ug, Lane 8: Lymph node (Rat) Lysate at 40 ug, Lane 9: Hela (Human) Cell Lysate at 30 ug, Lane 10: SW480 (Human) Cell Lysate at 30 ug, Lane 11: MCF-7 (Human) Cell Lysate at 30 ug, Lane 12: NIH/3T3 (Mouse) Cell Lysate at 30 ug, Lane 13: A549 (Human) Cell Lysate at 30 ug, Lane 14: A431 (Human) Cell Lysate at 30 ug, Primary: Anti-ERK1/2 (orb704524) at 1/1000 dilution, Secondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilution, Predicted band size: 44/42 kD, Observed band size: 44/42 kD.

Tissue/Cell: 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 (ERK1/2) monoclonal Antibody, Unconjugated (orb704524) 1:100, 90 minutes at 37°C, followed by a FITC conjugated Goat Anti-Rabbit IgG antibody at 37°C for 90 minutes, DAPI (blue) was used to stain the cell nuclei.