AKT1 Recombinant Rabbit Monoclonal Antibody [ST05-09]
cat.: ET1609-47
| Product Type: | Recombinant Rabbit monoclonal IgG, primary antibodies |
|---|---|
| Species reactivity: | Human, Mouse, Rat |
| Applications: | WB, IHC-P, IP, FC, IHC-Fr, IF-Cell |
| Clonality: | Monoclonal |
| Clone number: | ST05-09 |
| Form: | Liquid |
| Storage condition: | Shipped at 4℃. Store at +4℃ short term (1-2 weeks). Store at -20℃ long term. |
| 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 peptide within C-terminal human AKT1. |
| Positive control: | A549 cell lysate, C6 cell lysate, PC-12 cell lysate, NIH/3T3 cell lysates, mouse brain tissue, mouse kidney tissue, human kidney tissue, mouse prostate tissue, Hela, mouse hippocampus tissue, mouse cerebral cortex tissue, MCF7, C6. |
| Subcellular location: | Cell membrane, Cytoplasm, Membrane, Nucleus. |
| Recommended Dilutions:
WB IHC-P FC IP IHC-Fr IF-Cell |
1:5,000-1:10,000 1:2,000-1:5,000 1:1,000 1-2μg/sample 1:200 1:100 |
| 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 AKT1 on different lysates with Rabbit anti-AKT1 antibody (ET1609-47) at 1/5,000 dilution. Lane 1: C6 (Rat glioma cell) cell lysate Lane 2: PC-12 (Rat pheochromocytoma cell (undifferentiated)) cell lysate Lysates/proteins at 15 µg/Lane. Exposure time: 43 seconds; ECL: K1801 HCT 116 is a negative control (PMID: 32344898). Blocking: 5% NFDM/TBST, 1 hour at room temperature Primary antibody: ET1609-47, 1/5,000 in 5% NFDM/TBST, overnight at 4 ℃ 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 |
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Fig2:
Western blot analysis of AKT1 on different lysates with Rabbit anti-AKT1 antibody (ET1609-47) at 1/5,000 dilution. Lane 1: A549 (Human lung adenocarcinoma cell) cell lysate Lane 2: NIH/3T3 (Mouse fibroblast) cell lysate Lysates/proteins at 10 µg/Lane. Exposure time: 1 minute; ECL: K1801 HCT 116 is a negative control (PMID: 32344898). Blocking: 5% NFDM/TBST, 1 hour at room temperature Primary antibody: ET1609-47, 1/5,000 in 5% NFDM/TBST, 2 hours at room temperature 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 |
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Fig3:
Immunohistochemical analysis of paraffin-embedded mouse brain tissue with Rabbit anti-AKT1 antibody (ET1609-47) at 1/5,000 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 (ET1609-47) at 1/5,000 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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Fig4:
Immunohistochemical analysis of paraffin-embedded mouse kidney tissue with Rabbit anti-AKT1 antibody (ET1609-47) at 1/5,000 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 (ET1609-47) at 1/5,000 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:
Application: IHC-Fr Species: Mouse Site: Hippocampus Sample: Frozen section Antibody concentration: 1/200 Antigen retrieval: The section was pre-treated using 1% SDS buffer (in PBS, pH 7.4) for 5 minutes at room temperature. |
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Fig6:
Application: IHC-Fr Species: Mouse Site: Cerebral cortex Sample: Frozen section Antibody concentration: 1/200 Antigen retrieval: The section was pre-treated using 1% SDS buffer (in PBS, pH 7.4) for 5 minutes at room temperature. |
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Fig7:
Immunocytochemistry analysis of MCF7 cells labeling AKT1 with Rabbit anti-AKT1 antibody (ET1609-47) 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-AKT1 antibody (ET1609-47) 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 (M1305-2, 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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Fig8:
Immunocytochemistry analysis of C6 cells labeling AKT1 with Rabbit anti-AKT1 antibody (ET1609-47) 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-AKT1 antibody (ET1609-47) 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 (M1305-2, 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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Fig9:
Flow cytometric analysis of MCF7 cells labeling AKT1. Cells were fixed and permeabilized. Then stained with the primary antibody (ET1609-47, 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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Fig10:
AKT1 was immunoprecipitated from 0.2 mg MCF7 cell lysate with ET1609-47 at 2 µg/10 µl beads. Western blot was performed from the immunoprecipitate using ET1609-47 at 1/1,000 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: MCF7 cell lysate (input) Lane 2: ET1609-47 IP in MCF7 cell lysate Lane 3: Rabbit IgG instead of ET1609-47 in MCF7 cell lysate Blocking/Dilution buffer: 5% NFDM/TBST Exposure time: 23 seconds; ECL: K1801 |
Note: All products are “FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE”.