Phospho-AKT1 (T450) Recombinant Rabbit Monoclonal Antibody [SD08-12]
cat.: ET1612-73
| Product Type: | Recombinant Rabbit monoclonal IgG, primary antibodies |
|---|---|
| Species reactivity: | Human, Mouse, Rat |
| Applications: | WB, IHC-P, IP |
| Clonality: | Monoclonal |
| Clone number: | SD08-12 |
| Form: | Liquid |
| Storage condition: | Store at +4℃ after thawing. Aliquot store at -20℃ or -80℃. 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: 56 kDa |
| Isotype: | IgG |
| Immunogen: | Synthetic phospho-peptide corresponding to residues surrounding Thr450 of human AKT1. |
| Positive control: | MCF-7 cell lysates, NIH-3T3 cell lysates, human colon carcinoma tissue, human breast carcinoma tissue, mouse lung tissue. |
| Subcellular location: | Cytoplasm, Nucleus, Cell membrane. |
| Recommended Dilutions:
WB IHC-P IP |
1:500-1:2,000 1:50-1:200 Use at an assay dependent concentration. |
| Uniprot #: | SwissProt: P31749 Human | P31750 Mouse | P47196 Rat |
| Alternative names: | AKT 1 AKT 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 |
Images
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Fig1: Western blot analysis of Phospho-AKT1 (T450) on MCF-7 cell lysates. Proteins were transferred to a PVDF membrane and blocked with 5% BSA in PBS for 1 hour at room temperature. The primary antibody (ET1612-73, 1/500) was used in 5% BSA at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1:5,000 dilution was used for 1 hour at room temperature. |
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Fig2:
Western blot analysis of Phospho-AKT1 (T450) on NIH-3T3 cell lysates. Lane 1: NIH-3T3 cells, whole cell lysate, 10ug/lane Lane 2: NIH-3T3 cells treated with 2.8ug/ul lambda-PP for 30 minutes, whole cell lysates, 10ug/lane All lanes : Anti-Phospho-AKT1 (T450) antibody (ET1612-73 ) at 1:500 dilution. Anti-GAPDH antibody (ET1601-4) at 1:10,000 dilution. Goat Anti-Rabbit IgG H&L (HRP) (HA1001) at 1/200,000 dilution. Predicted band size: 56 kDa Observed band size: 60 kDa Blocking and diluting buffer: 5% BSA. Exposure time: 2 minutes |
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Fig3:
Western blot analysis of Phospho-AKT1 (T450) on MCF-7 cell lysates. Lane 1: MCF-7 cells, whole cell lysate, 10 ug/lane. Lane 2: MCF-7 cells treated with 2.8 μg/ul lambda-PP for 30 minutes, whole cell lysates, 10 ug/lane. Proteins were transferred to a PVDF membrane and blocked with 5% BSA in PBS for 1 hour at room temperature. The primary antibody Anti-Phospho-AKT1 (T450) antibody (ET1612-73, 1/1,000) , Anti-AKT1 antibody (ET1609-47, 1/5,000) and Anti-GAPDH antibody (ET1601-4, 1/10,000)was used in 5% BSA at room temperature for 2 hours. Goat Anti-Rabbit IgG H&L (HRP) Secondary Antibody (HA1001) at 1:200,000 dilution was used for 1 hour at room temperature. Predicted band size: 56 kDa Observed band size: 60 kDa Exposure time: 2 minutes |
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Fig4:
Immunohistochemical analysis of paraffin-embedded human colon carcinoma tissue with Rabbit anti-Phospho-AKT1 (T450) antibody (ET1612-73) at 1/50 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 (ET1612-73) at 1/50 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. |
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Fig5:
Immunohistochemical analysis of paraffin-embedded human breast carcinoma tissue with Rabbit anti-Phospho-AKT1 (T450) antibody (ET1612-73) at 1/50 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 (ET1612-73) at 1/50 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. |
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Fig6:
Immunohistochemical analysis of paraffin-embedded mouse lung tissue with Rabbit anti-Phospho-AKT1 (T450) antibody (ET1612-73) at 1/200 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 (ET1612-73) at 1/200 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”.