Anti-eIF4EBP1 (phospho T37) antibody [EPR28384-73]

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-eIF4EBP1 (phospho T37) antibody [EPR28384-73] (AB322664)

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized EIF4EBP1 KO HAP1 (EIF4EBP1 knockout human chronic myelogenous leukemia near-haploid cell) cells labelling eIF4EBP1 (phospho T37) with ab322664 at 1/50 (10.22 ug/ml) dilution, followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 2ug/ml dilution (Green).

Confocal image showing cytoplasmic and nuclear staining in parental HAP1 cell line (shown in green), the signal decreased after LY294002 (50 uM) for 2 h. The counterstain was observed in magenta. Nuclear DNA was labelled with DAPI (shown in blue). Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).

ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor® 594) was used to counterstain tubulin at 1/200 2.5ug/ml dilution (Magenta). The Nuclear counterstain was DAPI (Blue).

Secondary antibody only control : Secondary antibody is ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/ 1000 2ug/ml dilution.

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-eIF4EBP1 (phospho T37) antibody [EPR28384-73] (AB322664)

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized F9 (mouse embryonal carcinoma epithelial cell) cells labelling eIF4EBP1 (phospho T37) with ab322664 at 1/50 (10.22 ug/ml) dilution, followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 2ug/ml dilution (Green).

Confocal image showing cytoplasmic and nuclear staining in F9 cell line (shown in green), the signal both decreased after LY294002 (50 uM) for 2h or phosphatase treatment at 37℃ for 2h. The counterstain was observed in magenta. Nuclear DNA was labelled with DAPI (shown in blue). Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).

ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor® 594) was used to counterstain tubulin at 1/ 200 2.5ug/ml dilution (Magenta). The Nuclear counterstain was DAPI (Blue).

Secondary antibody only control : Secondary antibody is ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/ 1000 2ug/ml dilution.

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-eIF4EBP1 (phospho T37) antibody [EPR28384-73] (AB322664)

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized LNCaP (human prostate carcinoma epithelial cell) cells labelling eIF4EBP1 (phospho T37) with ab322664 at 1/50 (10.22 ug/ml) dilution, followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/ 1000 2ug/ml dilution (Green).

Confocal image showing cytoplasmic and nuclear staining in LNCaP cell line (shown in green), the signal both decreased after LY294002 (50 uM) for 2h or phosphatase treatment at 37℃ for 2h. The counterstain was observed in magenta. Nuclear DNA was labelled with DAPI (shown in blue). Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).

ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor® 594) was used to counterstain tubulin at 1/ 200 2.5ug/ml dilution (Magenta). The Nuclear counterstain was DAPI (Blue).

Secondary antibody only control : Secondary antibody is ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/ 1000 2ug/ml dilution.

• Dot

Supplier Data

Dot Blot - Anti-eIF4EBP1 (phospho T37) antibody [EPR28384-73] (AB322664)

Dot blot analysis of eIF4EBP1 (phospho T37) using ab322664 at 1000 (0.511 ug/ml) followed by a Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (ab97051) at 1 : 100,000 dilution.

Lane1 : Human eIF4EBP1 (phospho T37) peptide
Lane2 : Human eIF4EBP1 non-phospho control peptide for lane 1
Lane3 : Human eIF4EBP1 (phospho T46) peptide
Lane4 : Human eIF4EBP1 (phospho T36) peptide
Lane5 : Human eIF4EBP2 (phospho T37) peptide
Lane6 : Human eIF4EBP3 (phospho T23) peptide

Blocking and diluting buffer and concentration : 5% NFDM/TBST.

When tested in dot blot this antibody recognizes eIF4EBP1 phosphorylated at Thr37 and does not cross-react with eIF4EBP1 phosphorylated at Thr46 or Thr36. Antibody may cross-react with eIF4EBP2 and eIF4EBP3 when phosphorylated at equivalent sites.

All lanes:

Dot Blot - Anti-eIF4EBP1 (phospho T37) antibody [EPR28384-73] (ab322664) at 1/1000 dilution

Lane 1:

Human eIF4EBP1 (phospho T37) peptide a

Lane 2:

Human eIF4EBP1 non-phospho control peptide for lane 1

Lane 3:

Human eIF4EBP1 (phospho T46) peptide b

Lane 4:

Human eIF4EBP1 (phospho T36) peptide c

Lane 5:

Human eIF4EBP2 (phospho T37) peptide d

Lane 6:

Human eIF4EBP3 (phospho T23) peptide e

Secondary

All lanes:

Dot Blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/100000 dilution

false

Exposure time: 180s