eIF4EBP1 (Thr37 + Thr46) Antibody

Paraformaldehyde-fixed, paraffin embedded mouse small intestine tissue; Antigen retrieval by boiling in sodium citrate buffer(pH6) for 15min; Block endogenous peroxidase by 3% hydrogen peroxide for 30 minutes; Blocking buffer (normal goat serum) at 37\u00b0C for 20min; Antibody incubation with Rabbit Anti-eIF4EBP1 Polyclonal Antibody, Unconjugated (bs-3019R) at 1:400 overnight at 4\u00b0C, followed by a conjugated secondary and DAB staining\r\nImage on the left is the same conditions, with the peptide incubated with the antibody prior to staining.

Mouse ovary and Rat ovary lysates probed with eIF4EBP1 (Thr37+Thr46) Polyclonal Antibody, unconjugated (bs-3019R) at 1:300 overnight at 4\u00b0C followed by a conjugated secondary antibody at 1:10000 for 90 minutes at 37\u00b0C.

HepG2 cells were\u00a0fixed with 4% PFA for 10min at room temperature,permeabilized with\u00a090% ice-cold methanol for 20 min at -20\u2103,\u00a0and incubated in 5% BSA blocking buffer\u00a0for 30 min at room temperature. Cells were then stained with phospho-eIF4EBP1 (Thr37 + Thr46) Antibody(bs-3019R)at 1:100 dilution\u00a0in blocking buffer and\u00a0incubated for 30 min at\u00a0room temperature,\u00a0washed twice with 2%BSA in PBS,\u00a0followed by\u00a0secondary antibody incubation\u00a0for 40 min\u00a0at\u00a0room temperature. Acquisitions of 20,000 events were performed.\u00a0Cells stained with primary antibody\u00a0(green), and isotype control (orange).

HepG2 cell; 4% Paraformaldehyde-fixed; Triton X-100 at room temperature for 20 min; Blocking buffer (normal goat serum, C-0005) at 37\u00b0C for 20 min; Antibody incubation with (phospho-eIF4EBP1 (Thr37 + Thr46)) polyclonal Antibody, Unconjugated (bs-3019R) 1:100, 90 minutes at 37\u00b0C; followed by a conjugated Goat Anti-Rabbit IgG antibody at 37\u00b0C for 90 minutes, DAPI (blue, C02-04002) was used to stain the cell nuclei.

Hela cell; 4% Paraformaldehyde-fixed; Triton X-100 at room temperature for 20 min; Blocking buffer (normal goat serum, C-0005) at 37\u00b0C for 20 min; Antibody incubation with (phospho-eIF4EBP1 (Thr37 + Thr46)) polyclonal Antibody, Unconjugated (bs-3019R) 1:100, 90 minutes at 37\u00b0C; followed by a conjugated Goat Anti-Rabbit IgG antibody at 37\u00b0C for 90 minutes, DAPI (blue, C02-04002) was used to stain the cell nuclei.