AKT1/2/3 Antibody (16E10) - BSA Free

Immunohistochemistry: AKT1/2/3 Antibody (16E10) [NBP3-26514] - Image of AKT1/2/3 Antibody (16E10) diluted at 1:50 and staining in paraffin-embedded human brain tissue performed. After dewaxing and hydration, antigen retrieval was mediated by high pressure in a citrate buffer (pH 6.0). Section was blocked with 10% normal goat serum 30min at RT. Then primary antibody (1% BSA) was incubated at 4°C overnight. The primary is detected by a Goat anti-rabbit polymer IgG labeled by HRP and visualized using 0.54% DAB.

Western Blot: AKT1/2/3 Antibody (16E10) [NBP3-26514] - Positive Western Blot detected in: SH-SY5Y whole cell lysate. All lanes: AKT1/2/3 Antibody at 1: 1000 Secondary: Goat polyclonal to rabbit IgG at 1/50000 dilution. Predicted band size: 56 kDa Observed band size: 56 kDa

Flow Cytometry: AKT1/2/3 Antibody (16E10) [NBP3-26514] - Overlay Peak curve showing Jurkat cells stained with AKT1/2/3 Antibody (16E10) (red line) at 1:50. The cells were fixed in 4% formaldehyde and permeated by 0.2% Triton X-100. Then 10% normal goat serum to block non-specific protein-protein interactions followed by the antibody (1ug/1*10^6 cells) for 45min at 4C. The secondary antibody used was FITC-conjugated Goat Anti-rabbit IgG (H+L) at 1:200 dilution for 35min at 4C. Control antibody (green line) was rabbit IgG (1ug/1*10^6 cells) used under the same conditions. Acquisition of >10,000 events was performed.

• Reactivity: Hu
• Applications: WB, ELISA, Flow, IHC
• Clone: 16E10
• Clonality: Monoclonal
• Host: Rabbit
• Conjugate: Unconjugated
• Format: BSA Free

AKT1/2/3 Antibody (16E10) - BSA Free Summary

• Additional Information: Recombinant monoclonal antibody expressed in HEK293F cells
• Immunogen: A synthesized peptide derived from Human AKT1/2/3 [UniProt P31749/P31751/Q9Y243]
• Isotype: IgG
• Clonality: Monoclonal
• Host: Rabbit
• Gene: AKT1
• Purity: Affinity purified

Applications/Dilutions

• Dilutions:
  • ELISA
  • Flow Cytometry 1:50-1:200
  • Immunohistochemistry 1:50-1:200
  • Western Blot 1:500-1:2000

Packaging, Storage & Formulations

• Storage: Store at -20 to -70C. Avoid freeze-thaw cycles.
• Buffer: PBS, pH 7.4, 150mM NaCl, and 50% glycerol
• Preservative: 0.02% Sodium Azide
• Purity: Affinity purified

Alternate Names for AKT1/2/3 Antibody (16E10) - BSA Free

• 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 serine/threonine-protein kinase
• RAC-ALPHA
• RAC-PK-alpha
• RACPKB-ALPHA
• v-akt murine thymoma viral oncogene homolog 1

Background

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.

Limitations

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.

Bioinformatics

• Gene Symbol: AKT1