PRAS40 Antibody (JE30-60)

Western Blot: PRAS40 Antibody (JE30-60) [NBP3-32850]

Western Blot: PRAS40 Antibody (JE30-60) [NBP3-32850] - Western blot analysis of PRAS40 on different lysates with Rabbit anti-PRAS40 antibody (NBP3-32850) at 1/1,000 dilution.

Lane 1: HeLa cell lysate
Lane 2: MCF7 cell lysate
Lane 3: NIH/3T3 cell lysate
Lane 4: PC-12 cell lysate

Lysates/proteins at 40 ug/Lane.

Predicted band size: 27 kDa
Observed band size: 40 kDa

Exposure time: 1 minute 40 seconds;

4-20% SDS-PAGE gel.

Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (NBP3-32850) at 1/1,000 dilution was used in 5% NFDM/TBST at 4C overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody at 1:50,000 dilution was used for 1 hour at room temperature.

Immunohistochemistry: PRAS40 Antibody (JE30-60) [NBP3-32850]

Immunohistochemistry: PRAS40 Antibody (JE30-60) [NBP3-32850] - Immunohistochemical analysis of paraffin-embedded human brain tissue with Rabbit anti-PRAS40 antibody (NBP3-32850) at 1/200 dilution.

The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (NBP3-32850) at 1/200 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

Immunocytochemistry/ Immunofluorescence: PRAS40 Antibody (JE30-60) [NBP3-32850]

Immunocytochemistry/ Immunofluorescence: PRAS40 Antibody (JE30-60) [NBP3-32850] - Immunocytochemistry analysis of HeLa cells labeling PRAS40 with Rabbit anti-PRAS40 antibody (NBP3-32850) at 1/100 dilution.

Cells were fixed in 4% paraformaldehyde for 20 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 5 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Rabbit anti-PRAS40 antibody (NBP3-32850) at 1/100 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI.

Beta tubulin (red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor™ 594) was used as the secondary antibody at 1/1,000 dilution.