P70 S6 Kinase beta Recombinant Rabbit Monoclonal Antibody [JE35-69]
cat.: HA721354
Product Type: Recombinant Rabbit monoclonal IgG, primary antibodies
Species reactivity: Human, Mouse
Applications: WB, IF-Cell, FC, IHC-P
Clonality: Monoclonal
Clone number: JE35-69
Form: Liquid
Storage condition: Store at +4℃ after thawing. Aliquot store at -20℃. Avoid repeated freeze / thaw cycles.
Storage buffer: 1*TBS (pH7.4), 0.05% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide.
Concentration: 1ug/ul
Purification: Protein A affinity purified.
Molecular weight: Predicted band size: 53 kDa
Isotype: IgG
Immunogen: Recombinant protein within Human P70 S6 Kinase beta aa 1-150 / 482.
Positive control: HeLa cell lysate, Jurkat cell lysate, HEK-293 cell lysate, mouse liver tissue lysate, mouse kidney tissue lysate, HeLa, MCF7, human colon tissue.
Subcellular location: Cytoplasm, Nucleus.
Recommended Dilutions:
  WB
  IF-Cell
  FC
  IHC-P
1:1,000
1:100
1:500-1:1,000
1:1,000
Uniprot #: SwissProt: Q9UBS0 Human | Q9Z1M4 Mouse
Alternative names: 70 kDa ribosomal protein S6 kinase 2 EC 2.7.11.1 KS6B2_HUMAN p70 beta p70 ribosomal S6 kinase beta p70 S6 kinase beta p70 S6K-beta p70 S6KB p70 S6Kbeta p70(S6K) beta p70-beta p70-S6K 2 P70S6K2 p70S6Kb Ribosomal protein S6 kinase 70kDa, polypeptide 2 Ribosomal protein S6 kinase B2 Ribosomal protein S6 kinase beta 2 Ribosomal protein S6 kinase beta-2 Rps6kb2 S6 kinase related kinase S6 kinase-related kinase S6K beta 2 S6K beta S6K-beta S6K-beta-2 S6K2 Serine/threonine protein kinase 14 beta Serine/threonine-protein kinase 14B SRK STK14B
Images
HA721354_1.jpg Fig1: 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.
HA721354_2.jpg Fig2: 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.
HA721354_3.jpg Fig3: 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.
HA721354_4.jpg Fig4: 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).
HA721354_5.jpg Fig5: 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.
Note: All products are “FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE”.