Anti-Phospho-AKT1-(Ser473) Antibody, clone 1O14 ZooMAb^® Rabbit Monoclonal

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ZooMAb^® antibodies represent an entirely new generation of recombinant monoclonal antibodies.Each ZooMAb^® antibody is manufactured using our proprietary recombinant expression system, purified to homogeneity, and precisely dispensed to produce robust and highly reproducible lot-to-lot consistency. Only top-performing clones are released for use by researchers. Each antibody is validated for high specificity and affinity across multiple applications, including its most commonly used application. ZooMAb^® antibodies are reliably available and ready to ship when you need them.

Clone 1O14 is a ZooMAb^® Rabbit recombinant monoclonal antibody that specifically detects AKT1 phosphorylated on serine 473.

KLH-conjugated linear peptide corresponding to 11 amino acids surrounding phosphoserine 473 from the C-terminal region of human AKT1.

Quality Control Testing

Evaluated by Western Blotting in lysate from NIH3T3 cells treated with platelet derived growth factor (PDGF).

Western Blotting Analysis (WB): A 1:10,000 dilution of this antibody detected AKT1 in lysate from NIH3T3 cells serum starved overnight and treated with PDGF (100 ng/mL; 20 min).

Tested applications

Western Blotting Analysis (WB): A 1:1,000 dilution of this antibody detected AKT1 in lysate from HEK293 cells serum starved overnight and treated with Insulin (500 nM; 10 min).

Peptide Inhibition Assay: Target band detection in a lysate from HEK293 cells treated with insulin was prevented by preblocking of a representative lot with the immunogen phosphopeptide, but not the corresponding non-phosphopeptide.

Affinity Binding Assay: A representative lot of this antibody bound AKT1 peptide with a KD of 1.1 x 10-8 in an affinity binding assay.

Immunohistochemistry (Paraffin) Analysis: A 1:100 dilution from a representative lot detected Phospho-AKT1-(Ser473) in human breast cancer and human melanoma tissue sections.

Note: Actual optimal working dilutions must be determined by end user as specimens, and experimental conditions may vary with the end user

RAC-alpha serine/threonine-protein kinase (UniProt: P31749; also known as EC:2.7.11.1, Protein kinase B, PKB, Protein kinase B alpha, PKB alpha, Proto-oncogene c-Akt, RAC-PK-alpha, Akt alpha, Akt1) is encoded by the AKT1 (also known as PKB, RAC) gene (Gene ID: 207) in human. AKT, a serine/threonine kinase, is a critical enzyme in several signal transduction pathways involved in cell proliferation, apoptosis, angiogenesis, and diabetes. In mammals three isoforms of Akt have been reported (AKT 1, 2, 3) that exhibit a high degree of homology, but differ slightly in the localization of their regulatory phosphorylation sites. AKT1 plays an important role in growth and anti-apoptosis; AKT2 acts primarily as a regulator of glucose metabolism. Although AKT3 does not contribute significantly to the maintenance of normal metabolism in tissues, it is essential for the attainment of normal organ size, probably influencing both cell size and number via mTOR activation. Each AKT isoform is composed of three functionally distinct regions: An N-terminal pleckstrin homology (PH) domain that provides a lipid-binding module to direct AKT to PIP2 and PIP3, a central catalytic domain, and a C-terminal hydrophobic motif. Activation of AKT involves growth factor binding to a receptor tyrosine kinase and activation of PI 3-kinase that phosphorylates membrane bound PIP2 to generate PIP3. The binding of PIP3 to the PH domain anchors AKT to the plasma membrane. AKT is activated following its phosphorylation at Thr308 on the kinase domain by PDK1 and on Ser473 in the hydrophobic motif by mTORC2. The principal role of AKT is to facilitate growth factor-mediated cell survival and to block apoptotic cell death. AKT achieves this by phosphorylating a variety of substrates, such as Bad, Caspase-9, Forkhead transcription factors, and GSK-3. This ZooMAb^® recombinant monoclonal antibody, generated by our propriety technology, offers significantly enhanced specificity, affinity, reproducibility, and stability over conventional monoclonals.

Purified recombinant rabbit monoclonal antibody IgG, lyophilized in PBS, 5% Trehalose, normal appearance a coarse or translucent resin. The PBS/trehalose components in the ZooMAb formulation can have the appearance of a semi-solid (bead like gel) after lyophilization. This is a normal phenomenon. Please follow the recommended reconstitution procedure in the data sheet to dissolve the semi-solid, bead-like, gel-appearing material. The resulting antibody solution is completely stable and functional as proven by full functional testing. Contains no biocide or preservatives, such as azide, or any animal by-products. Larger pack sizes provided as multiples of 25 μL.

300 μg/mL after reconstitution at 25 μL per vial. Please refer to guidance on suggested starting dilutions and/or titers per application and sample type.

Recommend storage of lyophilized product at 2-8°C; Before reconstitution, micro-centrifuge vials briefly to spin down material to bottom of the vial; Reconstitute each vial by adding 25 μL of filtered lab grade water or PBS; Reconstituted antibodies can be stored at 2-8°C, or -20°C for long term storage. Avoid repeated freeze-thaws.

ZooMAb is a registered trademark of Merck KGaA, Darmstadt, Germany

Unless otherwise stated in our catalog or other company documentation accompanying the product(s), our products are intended for research use only and are not to be used for any other purpose, which includes but is not limited to, unauthorized commercial uses, in vitro diagnostic uses, ex vivo or in vivo therapeutic uses or any type of consumption or application to humans or animals.