Phospho-eIF4E (Ser209) Rabbit Polyclonal Antibody

• Catalog Number: orb13390
• Category: Antibodies
• Description: Phospho-eIF4E (Ser209) Rabbit Polyclonal Antibody
• Species/Host: Rabbit
• Clonality: Polyclonal
• Tested applications: FC, IF, IHC-Fr, IHC-P, WB
• Predicted Reactivity: Bovine, Canine, Equine, Gallus, Porcine, Rabbit
• Reactivity: Human, Mouse, Rat, Zebrafish
• Isotype: IgG
• Immunogen: KLH conjugated Synthesised phosphopeptide derived from human eIF4E around the phosphorylation site of Ser209 SG(p-S)TT
• 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=0.2ug/test
• Form/Appearance: Liquid
• Conjugation: Unconjugated
• MW: 24 kDa
• Target: EIF4E
• UniProt ID: P06730
• RRID: AB_10751395
• 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: eIF4E(phospho S209); eIF4E(phospho Ser209); p-eIF4
• Note: For research use only
• Expiration Date: 12 months from date of receipt.

Blank control: Molt-4. Primary Antibody (green line): Rabbit Anti-phospho-eIF4E (Ser209) antibody (orb13390), Dilution: 0.2 µg/10^6 cells, Isotype Control Antibody (orange line): Rabbit IgG. Secondary Antibody: Goat anti-rabbit IgG-PE, Dilution: 0.2 µg/Test. Protocol, The cells were fixed with 4% PFA (10 min at room temperature) and then permeabilized with 20% PBST 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 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 (phospho-eIF4E (Ser209)) Polyclonal Antibody, Unconjugated (orb13390) at 1:400 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 (phospho-eIF4E (Ser209)) Polyclonal Antibody, Unconjugated (orb13390) at 1:400 overnight at 4°C, followed by operating according to SP Kit (Rabbit) instructionsand DAB staining.

Paraformaldehyde-fixed, paraffin embedded (rat testis), 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-eIF4E (Ser209)) Polyclonal Antibody, Unconjugated (orb13390) at 1:200 overnight at 4°C, followed by operating according to SP Kit (Rabbit) instructionsand DAB staining.

Sample: NIH/3T3 (Mouse) Cell Lysate at 30 ug, Primary: Anti-phospho-eIF4E (Ser209) (orb13390) at 1/500 dilution, Secondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilution, Predicted band size: 24 kD, Observed band size: 28 kD.