AKT1/2/3 Antibody (JE75-09)

Novus Biologicals, part of Bio-Techne | Catalog # NBP3-32017

Recombinant Monoclonal Antibody
Novus Biologicals, part of Bio-Techne
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NBP3-32017
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Key Product Details

Species Reactivity

Human, Mouse

Applications

Flow Cytometry, Immunocytochemistry/ Immunofluorescence, Western Blot

Label

Unconjugated

Antibody Source

Recombinant Monoclonal Rabbit IgG Clone # JE75-09

Concentration

1 mg/ml

View all AKT1/2/3 Antibodies »

Product Specifications

Immunogen

Recombinant protein within Human AKT1/2/3 aa 231-480 / 480. (Uniprot: P31749)

Localization

Cytoplasm, Nucleus, Cell membrane.

Clonality

Monoclonal

Host

Rabbit

Isotype

IgG

Theoretical MW

56 kDa.
Disclaimer note: The observed molecular weight of the protein may vary from the listed predicted molecular weight due to post translational modifications, post translation cleavages, relative charges, and other experimental factors.

Scientific Data Images for AKT1/2/3 Antibody (JE75-09)

AKT1/2/3 Antibody (JE75-09)

Immunocytochemistry/ Immunofluorescence: AKT1/2/3 Antibody (JE75-09) [NBP3-32017]

Immunocytochemistry/ Immunofluorescence: AKT1/2/3 Antibody (JE75-09) [NBP3-32017] - Immunocytochemistry analysis of MCF7 cells labeling AKT1/2/3 with Rabbit anti-AKT1/2/3 antibody (NBP3-32017) at 1/100 dilution.

Cells were fixed in 100% precooled methanol 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-AKT1/2/3 antibody (NBP3-32017) at 1/100 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488) 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 (red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor™ 594) was used as the secondary antibody at 1/1,000 dilution.
AKT1/2/3 Antibody (JE75-09)

Flow Cytometry: AKT1/2/3 Antibody (JE75-09) [NBP3-32017]

Flow Cytometry: AKT1/2/3 Antibody (JE75-09) [NBP3-32017] - Flow cytometric analysis of MCF7 cells labeling AKT1/2/3.

Cells were fixed and permeabilized. Then stained with the primary antibody (NBP3-32017, 1μg/mL) (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 at 1/1,000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black).
AKT1/2/3 Antibody (JE75-09)

Western Blot: AKT1/2/3 Antibody (JE75-09) [NBP3-32017]

Western Blot: AKT1/2/3 Antibody (JE75-09) [NBP3-32017] - Western blot analysis of AKT1/2/3 on different lysates with Rabbit anti-AKT1/2/3 antibody (NBP3-32017) at 1/2,000 dilution.
Lane 1: HeLa cell lysate
Lane 2: MCF7 cell lysate
Lane 3: A549 cell lysate
Lane 4: U-2 OS cell lysate
Lane 5: NIH/3T3 cell lysate

Lysates/proteins at 15 µg/Lane.

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

Exposure time: 30 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 at 1/2,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody at 1/50,000 dilution was used for 1 hour at room temperature.

Applications for AKT1/2/3 Antibody (JE75-09)

Application
Recommended Usage

Flow Cytometry

1:1000

Immunocytochemistry/ Immunofluorescence

1:100

Western Blot

1:2000
Please Note: Optimal dilutions of this antibody should be experimentally determined.

Formulation, Preparation, and Storage

Purification

Protein A purified

Formulation

1*TBS (pH7.4), 0.05% BSA and 40% Glycerol

Preservative

0.05% Sodium Azide

Concentration

1 mg/ml

Shipping

The product is shipped with polar packs. Upon receipt, store it immediately at the temperature recommended below.

Stability & Storage

Store at 4C short term. Aliquot and store at -20C long term. Avoid freeze-thaw cycles.

Background: AKT1/2/3

AKT, also known as protein kinase B (PKB), is a 57 kDa serine/threonine protein kinase. There are three mammalian isoforms of Akt: AKT1 (PKB alpha), AKT2 (PKB beta) and AKT3 (PKB gamma) with AKT2 and AKT3 being approximately 82% identical with the AKT1 isoform. Each isoform has a pleckstrin homology (PH)domain, a kinase domain and a carboxy terminal regulatory domain. AKT was originally cloned from the retrovirus AKT8, and is a key regulator of many signal transduction pathways. Its tight control over cell proliferation and cell viability are manifold; overexpression or inappropriate activation of AKT has been seen in many types of cancer. AKT mediates many of the downstream events of phosphatidylinositol 3 kinase (a lipid kinase activated by growth factors, cytokines and insulin). PI3 kinase recruits AKT to the membrane, where it is activated by PDK1 phosphorylation. Once phosphorylated, AKT dissociates from the membrane and phosphorylates targets in the cytoplasm and the cell nucleus. AKT has two main roles: (i) inhibition of apoptosis; (ii) promotion of proliferation. AKT has been shown to play a role in such metabolic processes as glucose transport, glycogen synthesis, glycolysis, and protein synthesis. It had also been shown to promote cell survival by inhibiting apoptosis through its ability to phosphoylate and inactivate several targets, including Bad, Forkhead transcription factors, and caspase 9. Activity of AKT has been associated with the phosphorylation of two sites: T308, in the activation loop of the kinase, and S473, at the carboxyl terminus. Phosphorylation of both sites contributes to AKT activity, however phosphorylation of T308 has been shown to be absolutely essential for AKT activation.

Alternate Names

AKT, EC 2.7.11, EC 2.7.11.1, PKBMGC99656, PRKBA, Protein kinase B, Proto-oncogene c-Akt, rac protein kinase alpha, RAC-ALPHA, RAC-alpha serine/threonine-protein kinase, RAC-PK-alpha, RACPKB-ALPHA, v-akt murine thymoma viral oncogene homolog 1

Gene Symbol

AKT1

Additional AKT1/2/3 Products

Product Documents for AKT1/2/3 Antibody (JE75-09)

Product Datasheets

Certificate of Analysis

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Product Specific Notices for AKT1/2/3 Antibody (JE75-09)

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Primary Antibodies are guaranteed for 1 year from date of receipt.

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