Phospho-AKT (T308) Recombinant Rabbit Monoclonal Antibody [PSH08-10]
cat.: HA722951
| Product Type: | Recombinant Rabbit monoclonal IgG, primary antibodies |
|---|---|
| Species reactivity: | Human |
| Applications: | WB, IF-Cell, FC |
| Clonality: | Monoclonal |
| Clone number: | PSH08-10 |
| Form: | Liquid |
| Storage condition: | Store at +4℃ after thawing. Aliquot store at -20℃. Avoid repeated freeze / thaw cycles. |
| 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: 56 kDa |
| Isotype: | IgG |
| Immunogen: | Synthetic phospho-peptide corresponding to residues surrounding Thr308 of Human AKT1. |
| Positive control: | Jurkat treated with 100nM Calyculin A for 30 minutes cell lysate, HeLa treated with 100ng/mL Calyculin A for 30 minutes cell lysate, 293T treated with 100nM Calyculin A for 15 minutes cell lysate, Jurkat cells treated with 100nM Calyculin A for 30 minutes. |
| Subcellular location: | Cytoplasm, Nucleus, Cell membrane. |
| Recommended Dilutions:
WB IF-Cell FC |
1:2,000 1:100 1:1,000 |
| Uniprot #: | SwissProt: P31749 Human |
| Alternative names: | AKT1 C AKT PKB PRKBA RAC alpha RAC alpha serine/threonine-protein kinase RAC RAC PK alpha v akt murine thymoma viral oncogene homolog 1 |
Images
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Fig1:
Western blot analysis of Phospho-AKT (T308) on different lysates with Rabbit anti-Phospho-AKT (T308) antibody (HA722951) at 1/2,000 dilution. Lane 1: Jurkat cell lysate Lane 2: Jurkat treated with 100nM Calyculin A for 30 minutes cell lysate Lane 3: HeLa cell lysate Lane 4: HeLa treated with 100ng/mL Calyculin A for 30 minutes cell lysate Lane 5: 293T cell lysate Lane 6: 293T treated with 100nM Calyculin A for 15 minutes cell lysate Lysates/proteins at 30 µg/Lane. Predicted band size: 56 kDa Observed band size: 56 kDa Exposure time: 3 minutes; 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 (HA722951) 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. |
|
Fig2:
Immunocytochemistry analysis of Jurkat cells treated with or without 100nM Calyculin A for 30 minutes labeling Phospho-AKT (T308) with Rabbit anti-Phospho-AKT (T308) antibody (HA722951) at 1/100 dilution. Cells were fixed in 4% paraformaldehyde for 20 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 5 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-Phospho-AKT (T308) antibody (HA722951) 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. |
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Fig3:
Flow cytometric analysis of Jurkat cells treated with or without 100nM Calyculin A for 30 minutes labeling Phospho-AKT (T308). Cells were fixed and permeabilized. Then stained with the primary antibody (HA722951, 1μg/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). |
Note: All products are “FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE”.