AKT3 Rabbit Polyclonal Antibody
cat.: ER1901-33
Product Type: Rabbit polyclonal IgG, primary antibodies
Species reactivity: Human, Mouse, Rat
Applications: WB, IF-Cell, IHC-P, FC
Clonality: Polyclonal
Form: Liquid
Storage condition: Store at +4℃ after thawing. Aliquot store at -20℃. Avoid repeated freeze / thaw cycles.
Storage buffer: 1*PBS (pH7.4), 0.2% BSA, 50% Glycerol. Preservative: 0.05% Sodium Azide.
Concentration: 1ug/ul
Purification: Immunogen affinity purified.
Molecular weight: Predicted band size: 56 kDa
Isotype: IgG
Immunogen: Recombinant protein within Human AKT3 aa 211-383 / 479.
Positive control: MCF7 cell lysate, Jurkat cell lysate, C6 cell lysate, rat heart tissue lysate, A549, EA.hy926, human thyroid tissue, human kidney tissue, mouse brain tissue.
Subcellular location: Nucleus, Cytoplasm, Membrane.
Recommended Dilutions:
  WB
  IF-Cell
  IHC-P
  FC
1:500-1:2,000
1:50-1:100
1:50-1:200
1:50-1:100
Uniprot #: SwissProt: Q9Y243 Human | Q9WUA6 Mouse | Q63484 Rat
Alternative names: Akt3 AKT3 kinase AKT3_HUMAN DKFZp434N0250 MPPH PKB gamma PKBG PRKBG Protein kinase Akt-3 Protein Kinase AKT3 Protein kinase B gamma RAC gamma RAC gamma serine/threonine protein kinase RAC-gamma serine/threonine-protein kinase RAC-PK-gamma RACPK Gamma Serine threonine protein kinase Akt 3 Serine threonine protein kinase Akt3 STK 2 STK-2 STK2 V akt murine thymoma viral oncogene homolog 3 (protein kinase B, gamma) V akt murine thymoma viral oncogene homolog 3 V akt murine thymoma viral oncogene homolog 3 protein kinase B gamma
Images
ER1901-33_1.jpg Fig1: Western blot analysis of AKT3 on different lysates with Rabbit anti-AKT3 antibody (ER1901-33) at 1/1,000 dilution.

Lane 1: MCF7 cell lysate
Lane 2: Jurkat cell lysate
Lane 3: C6 cell lysate
Lane 4: Rat heart tissue lysate

Lysates/proteins at 20 µg/Lane.

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

Exposure time: 18 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 (ER1901-33) at 1/1,000 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.
ER1901-33_2.jpg Fig2: ICC staining of AKT3 in A549 cells (green). Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature and blocked with 1% Blocker BSA for 15 minutes at room temperature. Cells were probed with the primary antibody (ER1901-33, 1/50) for 1 hour at room temperature, washed with PBS. Alexa Fluor®488 Goat anti-Rabbit IgG was used as the secondary antibody at 1/1,000 dilution. The nuclear counter stain is DAPI (blue).
ER1901-33_3.jpg Fig3: ICC staining of AKT3 in EA.hy926 cells (green). Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature and blocked with 1% Blocker BSA for 15 minutes at room temperature. Cells were probed with the primary antibody (ER1901-33, 1/50) for 1 hour at room temperature, washed with PBS. Alexa Fluor®488 Goat anti-Rabbit IgG was used as the secondary antibody at 1/1,000 dilution. The nuclear counter stain is DAPI (blue).
ER1901-33_4.jpg Fig4: Immunohistochemical analysis of paraffin-embedded human thyroid tissue using anti-AKT3 antibody. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) 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 (ER1901-33, 1/200) 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.
ER1901-33_5.jpg Fig5: Immunohistochemical analysis of paraffin-embedded human kidney tissue using anti-AKT3 antibody. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) 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 (ER1901-33, 1/200) 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.
ER1901-33_6.jpg Fig6: Immunohistochemical analysis of paraffin-embedded mouse brain tissue using anti-AKT3 antibody. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) 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 (ER1901-33, 1/200) 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.
ER1901-33_7.jpg Fig7: Flow cytometric analysis of AKT3 was done on EA.hy926 cells. The cells were fixed, permeabilized and stained with the primary antibody (ER1901-33, 1/50) (red). 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; black).
Note: All products are “FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE”.