Phospho-AKT (S473) Recombinant Rabbit Monoclonal Antibody [PSH24-43]
cat.: HA751941
| Product Type: | Recombinant Rabbit monoclonal IgG, primary antibodies |
|---|---|
| Species reactivity: | Human, Mouse, Rat |
| Applications: | WB, IP |
| Clonality: | Monoclonal |
| Clone number: | PSH24-43 |
| Form: | Liquid |
| Storage condition: | Store at +4℃ after thawing. Aliquot store at -20℃. Avoid repeated freeze / thaw cycles. |
| Storage buffer: | 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 Ser473 of human Akt1. |
| Positive control: | 293T (Human embryonic kidney cells) cell lysate, 293T treated with 100 nM Calyculin A for 15 minutes cell lysate, C2C12 treated with 100 nM Insulin for 30 minutes cell lysate, C6 (Rat glioma cell) cell lysate, C6 treated with 50 ng/mL Calyculin A for 45 minutes cell lysate. |
| Subcellular location: | Cytoplasm, Nucleus, Cell membrane. |
| Recommended Dilutions:
WB IP |
1:1,000-1:2,000 1-2μg/sample |
| Uniprot #: | SwissProt: P31749 Human | P31751 Human | Q9Y243 Human | P31750 Mouse | Q60823 Mouse | Q9WUA6 Mouse | P47196 Rat | P47197 Rat | Q63484 Rat |
| Alternative names: | AKT1 AKT1_HUMAN MGC99656 PKB PKB-ALPHA PRKBA Protein Kinase B Alpha Protein kinase B Proto-oncogene c-Akt RAC Alpha RAC RAC-alpha serine/threonine-protein kinase RAC-PK-alpha akt s473 |
Images
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Fig1:
Western blot analysis of Phospho-AKT (S473) on different lysates with Rabbit anti-Phospho-AKT (S473) antibody (HA751941) at 1/1,000 dilution.
Lane 1: 293T (Human embryonic kidney cells) cell lysate Lane 2: 293T treated with 100 nM Calyculin A for 15 minutes cell lysate Lane 3: C2C12 (Mouse myoblasts) cell lysate Lane 4: C2C12 treated with 100 nM Insulin for 30 minutes cell lysate Lysates/proteins at 15 µg/Lane. Exposure time: 20 seconds; ECL: K1801 Blocking: 5% NFDM/TBST, 1 hour at room temperature Primary antibody: HA751941, 1/1,000 in primary antibody dilution buffer (K1803), overnight at 4 °C Secondary antibody: Goat anti-Rabbit IgG-HRP (HA1001), 1/50,000 in 5% NFDM/TBST, 1 hour at room temperature Predicted band size: 56 kDa Observed band size: 56 kDa |
|
Fig2:
Western blot analysis of Phospho-AKT (S473) on different lysates with Rabbit anti-Phospho-AKT (S473) antibody (HA751941) at 1/2,000 dilution.
Lane 1: C6 (Rat glioma cell) cell lysate Lane 2: C6 treated with 50 ng/mL Calyculin A for 45 minutes cell lysate Lysates/proteins at 10 µg/Lane. Exposure time: 17 seconds; ECL: K1801 Blocking: 5% NFDM/TBST, 1 hour at room temperature Primary antibody: HA751941, 1/2,000 in primary antibody dilution buffer (K1803), overnight at 4 °C Secondary antibody: Goat anti-Rabbit IgG-HRP (HA1001), 1/50,000 in 5% NFDM/TBST, 1 hour at room temperature Predicted band size: 56 kDa Observed band size: 56 kDa |
|
Fig3:
Immunoprecipitation (IP)
Phospho-AKT (S473) was immunoprecipitated in 0.2 mg NIH/3T3 (Mouse fibroblast) treated with 100 nM Calyculin A for 30 minutes cell lysate with HA751941 at 2 µg/10 µl beads. Western blot was performed from the immunoprecipitate using HA751941 at 1/5,000 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: NIH/3T3 treated with Calyculin A cell lysate (input) Lane 2: HA751941 IP in NIH/3T3 treated with Calyculin A cell lysate Lane 3: Rabbit IgG instead of HA751941 in NIH/3T3 treated with Calyculin A cell lysate Exposure time: 11 seconds Blocking: 5% NFDM/TBST, 1 hour at room temperature Primary dilution: HA751941, 1/2,000 in primary antibody dilution buffer (K1803), 2 hours at room temperature Predicted band size: 56 kDa Observed band size: 56 kDa |
Note: All products are “FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE”.