P70 S6 Kinase beta Recombinant Rabbit Monoclonal Antibody [JE35-69]

Western blot analysis of P70 S6 Kinase beta on different lysates with Rabbit anti-P70 S6 Kinase beta antibody (HA721354) at 1/1,000 dilution.

Lane 1: HeLa cell lysate, 20 µg/Lane
Lane 2: Jurkat cell lysate, 20 µg/Lane
Lane 3: HEK-293 cell lysate, 20 µg/Lane
Lane 4: Mouse liver tissue lysate, 40 µg/Lane
Lane 5: Mouse kidney tissue lysate, 40 µg/Lane

Predicted band size: 53 kDa
Observed band size: 53 kDa

Exposure time: 1 minute;

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 (HA721354) at 1/1,000 dilution was used in 5% NFDM/TBST at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1:100,000 dilution was used for 1 hour at room temperature.

☑ Knockdown (KD)

Western blot analysis of P70 S6 Kinase beta on different lysates with Rabbit anti-P70 S6 Kinase beta antibody (HA721354) at 1/1,000 dilution.

Lane 1: A549-si NT cell lysate
Lane 2: A549-si P70 S6 Kinase beta cell lysate

Lysates/proteins at 10 µg/Lane.

Predicted band size: 53 kDa
Observed band size: 53 kDa

Exposure time: 45 seconds; ECL: K1801;

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 (HA721354) at 1/1,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature.

Immunocytochemistry analysis of HeLa cells labeling P70 S6 Kinase beta with Rabbit anti-P70 S6 Kinase beta antibody (HA721354) at 1/100 dilution.

Cells were fixed in 4% paraformaldehyde for 10 minutes at 37 ℃, permeabilized with 0.05% Triton X-100 in PBS for 20 minutes, and then blocked with 2% negative goat serum for 30 minutes at room temperature. Cells were then incubated with Rabbit anti-P70 S6 Kinase beta antibody (HA721354) at 1/100 dilution in 2% negative goat serum overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) was used as the secondary antibody at 1/1,000 dilution. Nuclear DNA was labelled in blue with DAPI.

Beta tubulin (M1305-2, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor™ 594, HA1126) was used as the secondary antibody at 1/1,000 dilution.

Flow cytometric analysis of MCF7 cells labeling P70 S6 Kinase beta.

Cells were fixed and permeabilized. Then stained with the primary antibody (HA721354, 1ug/ml) (red) compared with Rabbit IgG Isotype Control (green). After incubation of the primary antibody at +4℃ for an hour, the cells were stained with a iFluor™ 488 conjugate-Goat anti-Rabbit IgG Secondary antibody (HA1121) at 1/1,000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black).

Immunohistochemical analysis of paraffin-embedded human colon tissue with Rabbit anti-P70 S6 Kinase beta antibody (HA721354) at 1/1,000 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 (HA721354) 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.