p70 S6 Kinase/S6K [p Thr389] Antibody (PSH02-23)

Immunohistochemistry: p70 S6 Kinase/S6K [p Thr389] Antibody (PSH02-23) [NBP3-32783]

Immunohistochemistry: p70 S6 Kinase/S6K [p Thr389] Antibody (PSH02-23) [NBP3-32783] - Immunohistochemical analysis of paraffin-embedded human breast carcinoma tissue with Rabbit anti-p70 S6 Kinase/S6K antibody (NBP3-32783) at 1/1,000 dilution.

The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) for 2 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 (NBP3-32783) at 1/1,000 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: p70 S6 Kinase/S6K [p Thr389] Antibody (PSH02-23) [NBP3-32783]

Immunocytochemistry/ Immunofluorescence: p70 S6 Kinase/S6K [p Thr389] Antibody (PSH02-23) [NBP3-32783] - Immunocytochemistry analysis of MCF7 cells treated with or without 20% FBS overnight then add 100nM Calyculin A for 30 minutes labeling p70 S6 Kinase/S6K with Rabbit anti-p70 S6 Kinase/S6K antibody (NBP3-32783) 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-p70 S6 Kinase/S6K antibody (NBP3-32783) 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.

Western Blot: p70 S6 Kinase/S6K [p Thr389] Antibody (PSH02-23) [NBP3-32783]

Western Blot: p70 S6 Kinase/S6K [p Thr389] Antibody (PSH02-23) [NBP3-32783] - Western blot analysis of p70 S6 Kinase/S6K on different lysates with Rabbit anti-p70 S6 Kinase/S6K antibody (NBP3-32783) at 1/2,000 dilution.
Lane 1: MCF7 cell lysate
Lane 2: MCF7 treated with 20% FBS overnight then add 100nM Calyculin A for 30 minutes cell lysate
Lane 3: NIH/3T3 cell lysate
Lane 4: NIH/3T3 treated with 20% FBS overnight then add 100nM Calyculin A for 30 minutes cell lysate

Lysates/proteins at 15 µg/Lane.

Predicted band size: 59 kDa
Observed band size: 70/85 kDa

Exposure time:
Lane 1-4 (left): 1 minute 2 seconds;
Lane 1-4 (right): 3 minutes;

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 at 1/2,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody at 1/50,000 dilution was used for 1 hour at room temperature.