p70 S6 Kinase Recombinant Rabbit Monoclonal Antibody [PSH13-50]
cat.: HA723539
| Product Type: | Recombinant Rabbit monoclonal IgG, primary antibodies |
|---|---|
| Species reactivity: | Human, Mouse, Rat, Monkey |
| Applications: | WB, IF-Cell, FC, IP |
| Clonality: | Monoclonal |
| Clone number: | PSH13-50 |
| Form: | Liquid |
| Storage condition: | Shipped at 4℃. Store at +4℃ short term (1-2 weeks). It is recommended to aliquot into single-use upon delivery. Store at -20℃ long term. |
| Storage buffer: | PBS (pH7.4), 0.1% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide. |
| Concentration: | 1ug/ul |
| Purification: | Protein A affinity purified. |
| Molecular weight: | Predicted band size: 59 kDa |
| Isotype: | IgG |
| Immunogen: | Recombinant protein within Human p70 S6 Kinase aa 1-75 and aa 454-525. |
| Positive control: | HeLa cell lysate, MCF7 cell lysate, 293T cell lysate, HL-60 cell lysate, Neuro-2a cell lysate, PC-12 cell lysate, COS-1 cell lysate, HeLa, Neuro-2a, C6. |
| Subcellular location: | Synapse, synaptosome, Mitochondrion outer membrane, Mitochondrion; Nucleus, Cytoplasm. |
| Recommended Dilutions:
WB IF-Cell FC IP |
1:2,000 1:100-1:250 1:1,000 1-2μg/sample |
| Uniprot #: | SwissProt: P23443 Human | Q8BSK8 Mouse | P67999 Rat |
| Alternative names: | 70 kDa ribosomal protein S6 kinase 1 KS6B1_HUMAN p70 alpha P70 beta 1 p70 ribosomal S6 kinase alpha p70 ribosomal S6 kinase beta 1 p70 S6 kinase alpha P70 S6 Kinase p70 S6 kinase, alpha 1 p70 S6 kinase, alpha 2 p70 S6K p70 S6K-alpha p70 S6KA p70(S6K) alpha p70(S6K)-alpha p70-alpha p70-S6K 1 p70-S6K P70S6K P70S6K1 p70S6Kb PS6K Ribosomal protein S6 kinase 70kDa polypeptide 1 Ribosomal protein S6 kinase beta 1 Ribosomal protein S6 kinase beta-1 Ribosomal protein S6 kinase I RPS6KB1 S6K S6K-beta-1 S6K1 Serine/threonine kinase 14 alpha Serine/threonine-protein kinase 14A STK14A |
Images
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Fig1:
Western blot analysis of p70 S6 Kinase on different lysates with Rabbit anti-p70 S6 Kinase antibody (HA723539) at 1/2,000 dilution. Lane 1: HeLa cell lysate Lane 2: MCF7 cell lysate Lane 3: 293T cell lysate Lane 4: HL-60 cell lysate Lane 5: Neuro-2a cell lysate Lane 6: PC-12 cell lysate Lane 7: COS-1 cell lysate Lysates/proteins at 20 µg/Lane. Predicted band size: 59 kDa Observed band size: 70 kDa Exposure time: 25 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 (HA723539) at 1/2,000 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. |
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Fig2:
Immunocytochemistry analysis of HeLa cells labeling p70 S6 Kinase with Rabbit anti-p70 S6 Kinase antibody (HA723539) at 1/250 dilution. Cells were fixed in 4% paraformaldehyde for 15 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 15 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 antibody (HA723539) at 1/250 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) 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 (HA601187, 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. |
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Fig3:
Immunocytochemistry analysis of Neuro-2a cells labeling p70 S6 Kinase with Rabbit anti-p70 S6 Kinase antibody (HA723539) at 1/100 dilution. Cells were fixed in 4% paraformaldehyde for 15 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 15 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 antibody (HA723539) at 1/100 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) 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 (HA601187, 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. |
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Fig4:
Immunocytochemistry analysis of C6 cells labeling p70 S6 Kinase with Rabbit anti-p70 S6 Kinase antibody (HA723539) at 1/100 dilution. Cells were fixed in 4% paraformaldehyde for 15 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 15 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 antibody (HA723539) at 1/100 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) 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 (HA601187, 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. |
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Fig5:
Flow cytometric analysis of Neuro-2a cells labeling p70 S6 Kinase. Cells were fixed and permeabilized. Then stained with the primary antibody (HA723539, 1/1,000) (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). |
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Fig6:
Flow cytometric analysis of C6 cells labeling p70 S6 Kinase. Cells were fixed and permeabilized. Then stained with the primary antibody (HA723539, 1/1,000) (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). |
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Fig7:
p70 S6 Kinase was immunoprecipitated from 0.2 mg HeLa cell lysate with HA723539 at 2 µg/10 µl beads. Western blot was performed from the immunoprecipitate using HA723539 at 1/5,000 dilution. HRP Conjugated Anti-Rabbit IgG for IP Nano-secondary antibody at 1/5,000 dilution was used for 1 hour at room temperature. Lane 1: HeLa cell lysate (input) Lane 2: HA723539 IP in HeLa cell lysate Lane 3: Rabbit IgG instead of HA723539 in HeLa cell lysate Blocking/Dilution buffer: 5% NFDM/TBST Exposure time: 3 minutes; ECL: K1802 |
Note: All products are “FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE”.