p70 S6 Kinase Recombinant Rabbit Monoclonal Antibody [JE03-20]

Western blot analysis of p70 S6 Kinase on different lysates with Rabbit anti-p70 S6 Kinase antibody (HA722520) at 1/1,000 dilution.

Lane 1: MCF7 cell lysate
Lane 2: A549 cell lysate
Lane 3: HepG2 cell lysate
Lane 4: NIH/3T3 cell lysate
Lane 5: C6 cell lysate
Lane 6: Mouse brain tissue lysate
Lane 7: Rat brain tissue lysate

Lysates/proteins at 20 µg/Lane.

Predicted band size: 59 kDa
Observed band size: 59 kDa

Exposure time: 10 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 (HA722520) 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.

Immunohistochemical analysis of paraffin-embedded human brain tissue with Rabbit anti-p70 S6 Kinase antibody (HA722520) 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 (HA722520) 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.

Immunohistochemical analysis of paraffin-embedded mouse brain tissue with Rabbit anti-p70 S6 Kinase antibody (HA722520) 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 (HA722520) 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.

Immunohistochemical analysis of paraffin-embedded rat brain tissue with Rabbit anti-p70 S6 Kinase antibody (HA722520) 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 (HA722520) 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.

p70 S6 Kinase was immunoprecipitated from 0.2 mg C6 cell lysate with HA722520 at 2 µg/10 µl beads. Western blot was performed from the immunoprecipitate using HA722520 at 1/1,000 dilution. Anti-Rabbit IgG for IP Nano-secondary antibody (NBI01H) at 1/5,000 dilution was used for 1 hour at room temperature.

Lane 1: C6 cell lysate (input)
Lane 2: HA722520 IP in C6 cell lysate
Lane 3: Rabbit IgG instead of HA722520 in C6 cell lysate

Blocking/Dilution buffer: 5% NFDM/TBST
Exposure time: 20 seconds; ECL: K1801