Phospho-AKT2 (S474) Recombinant Rabbit Monoclonal Antibody [PSH09-46]
cat.: HA751293
| Product Type: | Recombinant Rabbit monoclonal IgG, primary antibodies |
|---|---|
| Species reactivity: | Human, Mouse, Rat |
| Applications: | WB, FC |
| Clonality: | Monoclonal |
| Clone number: | PSH09-46 |
| Form: | Liquid |
| Storage condition: | Store at +4℃ after thawing. Aliquot store at -20℃. Avoid repeated freeze / thaw cycles. |
| Storage buffer: | 1*PBS (pH7.4). |
| Concentration: | 1ug/ul |
| Purification: | Protein A affinity purified. |
| Molecular weight: | Predicted band size: 56 kDa |
| Isotype: | IgG |
| Immunogen: | Synthetic phospho-peptide corresponding to residues surrounding Ser474 of human AKT2. |
| Positive control: | MCF7 cell lysate, MCF7 treated with 50ng/mL Calyculin A for 45 minutes cell lysate, HEK-293 cell lysate, HEK-293 treated with 50μM LY294002 for 6 hours cell lysate, NIH/3T3 treated with 100ng/mL PDGF for 1 hour cell lysate, C6 cell lysate, C6 treated with 100ng/mL Calyculin A for 1 hour cell lysate, MCF7 cells treated with 50ng/mL Calyculin A for 45 minutes, C6 cells treated with 100ng/mL Calyculin A for 1 hours. |
| Subcellular location: | Cytoplasm, Nucleus, Cell membrane, Early endosome. |
| Recommended Dilutions:
WB FC |
1:2,000 1:1,000 |
| Uniprot #: | SwissProt: P31751 Human | Q60823 Mouse | P47197 Rat |
| Alternative names: | AKT Akt2 AKT2_HUMAN HIHGHH Murine thymoma viral (v-akt) homolog 2 murine thymoma viral (v-akt) homolog-2 Oncogene AKT2 protein kinase B beta PKB PKB beta PKBB PKBBETA PRKBB Protein kinase Akt 2 Protein kinase Akt-2 Protein kinase B beta RAC beta rac protein kinase beta RAC-BETA RAC-beta serine/threonine-protein kinase RAC-PK-beta RACbeta v akt murine thymoma viral oncogene homolog 2 V-AKT murine thymoma viral oncogene homolog 2 |
Images
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Fig1:
Western blot analysis of Phospho-AKT2 (S474) on different lysates with Rabbit anti-Phospho-AKT2 (S474) antibody (HA751293) at 1/2,000 dilution. Lane 1: MCF7 cell lysate Lane 2: MCF7 treated with 50ng/mL Calyculin A for 45 minutes cell lysate Lane 3: HEK-293 cell lysate Lane 4: HEK-293 treated with 50μM LY294002 for 6 hours cell lysate Lane 5: NIH/3T3 cell lysate Lane 6: NIH/3T3 treated with 100ng/mL PDGF for 1 hour cell lysate Lane 7: C6 cell lysate Lane 8: C6 treated with 100ng/mL Calyculin A for 1 hour cell lysate Lysates/proteins at 30 µg/Lane. Predicted band size: 56 kDa Observed band size: 56 kDa Exposure time: Lane 1-6: 3 minutes; Lane 7-8: 12 seconds; ECL: K1802; 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 (HA751293) 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:
Flow cytometric analysis of MCF7 cells (left) / MCF7 cells treated with 50ng/mL Calyculin A for 45 minutes (right) labeling Phospho-AKT2 (S474). Cells were fixed and permeabilized. Then stained with the primary antibody (HA751293, 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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Fig3:
Flow cytometric analysis of C6 cells (left) / C6 cells treated with 100ng/mL Calyculin A for 1 hours (right) labeling Phospho-AKT2 (S474). Cells were fixed and permeabilized. Then stained with the primary antibody (HA751293, 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). |
Note: All products are “FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE”.