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
Clonality: Monoclonal
Clone number: ST05-09
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 peptide within C-terminal human AKT1.
Positive control: HeLa cell lysate, A549 cell lysate, C6 cell lysate, PC-12 cell lysate, SH-SY5Y cell lysates, 商品名 was done on Hela.
Subcellular location: Cell membrane, Cytoplasm, Membrane, Nucleus.
Recommended Dilutions:
  WB
  IHC-P
  FC
  IP
  IHC-Fr

1:2,000-1:5,000
1:50-1:200
1:50-1:100
Use at an assay dependent concentration.
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
ET1609-47_1.jpg Fig1: Western blot analysis of AKT1 on different lysates with Rabbit anti-AKT1 antibody (ET1609-47) at 1/5,000 dilution.

Lane 1: HeLa cell lysate
Lane 2: A549 cell lysate
Lane 3: C6 cell lysate
Lane 4: PC-12 cell lysate

Lysates/proteins at 15 µg/Lane.

Predicted band size: 56 kDa
Observed band size: 56 kDa

Exposure time: 43 seconds;

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 (ET1609-47) at 1/5,000 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.
ET1609-47_2.jpg Fig2: Western blot analysis of AKT1 on SH-SY5Y cell lysates with Rabbit anti-AKT1 antibody (ET1609-47) at 1/500 dilution.

Lysates/proteins at 10 µg/Lane.

Predicted band size: 56 kDa
Observed band size: 56 kDa

Exposure time: 2 minutes;

10% 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 (ET1609-47) at 1/500 dilution was used in 5% NFDM/TBST at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1:300,000 dilution was used for 1 hour at room temperature.
ET1609-47_3.jpg Fig3: Immunohistochemical analysis of paraffin-embedded human kidney tissue using anti-AKT1 antibody.

The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 8.0-8.4) for 20 minutes.The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1609-47, 1/50) for 30 minutes 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.
ET1609-47_4.jpg Fig4: Immunohistochemical analysis of paraffin-embedded mouse brain tissue using anti-AKT1 antibody.

The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 8.0-8.4) for 20 minutes.The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1609-47, 1/50) for 30 minutes 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.
ET1609-47_5.jpg Fig5: Immunohistochemical analysis of paraffin-embedded mouse prostate tissue using anti-AKT1 antibody.

The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 8.0-8.4) for 20 minutes.The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1609-47, 1/50) for 30 minutes 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.
ET1609-47_6.jpg Fig6: Immunohistochemical analysis of paraffin-embedded mouse kidney tissue using anti-AKT1 antibody.

The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 8.0-8.4) for 20 minutes.The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1609-47, 1/50) for 30 minutes 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.
ET1609-47_7.jpg Fig7: Flow cytometric analysis of AKT1 was done on Hela cells. The cells were fixed, permeabilized and stained with the primary antibody (ET1609-47, 1/50) (blue). After incubation of the primary antibody at room temperature for an hour, the cells were stained with a Alexa Fluor 488-conjugated Goat anti-Rabbit IgG Secondary antibody at 1/1000 dilution for 30 minutes.Unlabelled sample was used as a control (cells without incubation with primary antibody; red).
ET1609-47_8.jpg Fig8: Immunofluorescence analysis of frozen mouse hippocampus tissue labeling AKT1 with Rabbit anti-AKT1 antibody (ET1609-47).

The tissues were blocked in 3% BSA for 30 minutes at room temperature, washed with PBS, and then probed with the primary antibody (ET1609-47, green) at 1/100 dilution overnight at 4℃, washed with PBS. Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) was used as the secondary antibody at 1/200 dilution. Nuclei were counterstained with DAPI (blue). Image acquisition was performed with KFBIO KF-FL-400 Scanner.
ET1609-47_9.jpg Fig9: Immunofluorescence analysis of frozen mouse cerebral cortex tissue labeling AKT1 with Rabbit anti-AKT1 antibody (ET1609-47).

The tissues were blocked in 3% BSA for 30 minutes at room temperature, washed with PBS, and then probed with the primary antibody (ET1609-47, green) at 1/100 dilution overnight at 4℃, washed with PBS. Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) was used as the secondary antibody at 1/200 dilution. Nuclei were counterstained with DAPI (blue). Image acquisition was performed with KFBIO KF-FL-400 Scanner.
Note: All products are “FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE”.