ERK1 + ERK2 Rabbit Polyclonal Antibody

• Catalog Number: orb10604
• Category: Antibodies
• Description: Rabbit Anti-ERK1 + ERK2 antibody
• Species/Host: Rabbit
• Clonality: Polyclonal
• Tested applications: FC, IF, IHC-Fr, IHC-P, WB
• Predicted Reactivity: Bovine, Canine, Equine, Gallus, Goat, Porcine, Rabbit, Sheep
• Reactivity: Human, Mouse, Rat
• Isotype: IgG
• Immunogen: KLH conjugated synthetic peptide derived from human ERK2 (301-358/358aa)
• Antibody Type: Primary Antibody
• Concentration: 1mg/ml
• Dilution range: WB=1:500-2000, IHC-P=1:100-500, IHC-F=1:100-500, IF=1:100-500, Flow-Cyt=1μg/Test
• Form/Appearance: Liquid
• Conjugation: Unconjugated
• MW: 42 kDa
• Target: MAPK3
• UniProt ID: P28482
• RRID: AB_10749244
• 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: ERK 1/2; ERK 1; ERK 2; ERK-2; ERK1; ERK2; ERT1; ER
• Note: For research use only
• Expiration Date: 12 months from date of receipt.

References

Yu, Wu et al. BEX4 upregulation alters Sertoli cell growth properties and protein expression profiles: An explanation for cadmium-induced testicular Sertoli cell injury J Biochem Mol Toxicol, 31, (2017)

PubMed: 28295929

Blank control: Hep G2 cells (blue). Primary Antibody: Rabbit Anti-ERK1 + ERK2 antibody (orb10604), Dilution: 1 µg in 100 µl 1X PBS containing 0.5% BSA, Isotype Control Antibody: Rabbit IgG (orange), used under the same conditions, Secondary Antibody: Goat anti-rabbit IgG-PE (white blue), Dilution: 1:200 in 1 X PBS containing 0.5% BSA. Protocol, The cells were fixed with 2% paraformaldehyde (10 min), then permeabilized with 90% ice-cold methanol for 30 min on ice. Primary antibody (orb10604, 1 µg/1x10^6 cells) were incubated for 30 min on the ice, followed by 1 X PBS containing 0.5% BSA + 1 0% goat serum (15 min) to block non-specific protein-protein interactions. Then the Goat Anti-rabbit IgG/PE antibody was added into the blocking buffer mentioned above to react with the primary antibody at 1/200 dilution for 30 min on ice. 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 + ERK2) Polyclonal Antibody, Unconjugated (orb10604) at 1:400 overnight at 4°C, followed by operating according to SP Kit (Rabbit) instructionsand DAB staining.

Sample: Brain (Rat) Lysate at 30 ug, Heart (Rat) lysate at 30 ug, Primary: Anti-ERK2/MAPK1 (orb10604) at 1/200 dilution, Secondary: HRP conjugated Goat-Anti-rabbit IgG (orb572747) at 1/3000 dilution, Predicted band size: 42 kD, Observed band size: 42 kD.

Sample: Lane 1: A431 (Human) Cell Lysate at 30 ug, Lane 2: MCF-7 (Human) Cell Lysate at 30 ug, Lane 3: Huvec (Human) Cell Lysate at 30 ug, Primary: Anti-ERK1 + ERK2 (orb10604) at 1/1000 dilution, Secondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilution, Predicted band size: 44/42 kD, Observed band size: 42/40 kD.

Sample: Lane 1: Cerebrum (Mouse) Lysate at 40 ug, Lane 2: Cerebrum (Rat) Lysate at 40 ug, Lane 3: Lymph node (Mouse) Lysate at 40 ug, Lane 4: Lymph node (Rat) Lysate at 40 ug, Primary: Anti-ERK1 + ERK2 (orb10604) at 1/1000 dilution, Secondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilution, Predicted band size: 44/42 kD, Observed band size: 40 kD.

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-ERK2/MAPK1 Polyclonal Antibody, Unconjugated (orb10604) 1:200, overnight at 4°C, followed by conjugation to the secondary antibody and DAB staining.