Anti-Phospho-eIF4EBP1 (Thr37+Thr46) Polyclonal Antibody is a Rabbit antibody targeting Phospho-eIF4EBP1 (Thr37+Thr46). Anti-Phospho-eIF4EBP1 (Thr37+Thr46) Polyclonal Antibody can be used in IHC-P,IHC-Fr,ICC/IF,IF,FCM.

IgG

Human (predicted:Mouse,Rat,Dog,Pig,Cow,Horse,Rabbit,Goat)

1. Tissue/cell: human lung carcinoma; 4% Paraformaldehyde-fixed and paraffin-embedded;
Antigen retrieval: citrate buffer (0.01 M, pH 6.0), Boiling bathing for 15 min; Block endogenous peroxidase by 3% Hydrogen peroxide for 30 min; Blocking buffer (normal goat serum) at 37℃ for 20 min;
Incubation: Anti-phospho-eIF4EBP1 (Thr37+Thr46) Polyclonal Antibody, Unconjugated (TMAB-10639) 1: 200, overnight at 4°C, followed by conjugation to the secondary antibody and DAB staining
2. Blank control (black line): HepG2.
Primary Antibody (green line): Rabbit Anti-phospho-eIF4EBP1 (Thr37+Thr46) antibody (TMAB-10639)
Dilution: 1 μg/Test;
Secondary Antibody (white blue line): Goat anti-rabbit IgG-AF488
Dilution: 0.5 μg/Test.
Isotype control (orange line): Normal Rabbit IgG
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℃, 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 20,000 events was performed.
3. HepG2 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-eIF4EBP1 (Thr37+Thr46)) polyclonal Antibody, Unconjugated (TMAB-10639) 1:100, 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.

IHC-P: 1:100-500; IHC-Fr: 1:100-500; ICC/IF: 1:100-500; IF: 1:100-500; FCM: 1μg/Test

Polyclonal

KLH conjugated Synthesised phosphopeptide: human 4EBP1 around the phosphorylation site of Thr37/46

Human

EIF4EBP1

Eukaryotic translation initiation factor 4E-binding protein 1

Regulates eIF4E activity by preventing its assembly into the eIF4F complex. Mediates the regulation of protein translation by hormones, growth factors and other stimuli that signal through the MAP kinase and mTORC1 pathways.