AKT1 antibody | AbD02147

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Human anti AKT1

Product Type
Monoclonal Antibody
Clone
AbD02147
Isotype
HuCAL Fab bivalent
Product CodeApplicationsDatasheetMSDSPack SizeList PriceQuantity
HCA003 E WB 0.1 mg
Human anti Human AKT1 antibody, clone AbD02147 recognizes AKT1, a cytoplasmic kinase which is expressed in the nucleus after activation by integrin-linked protein kinase 1 (ILK1). Clone AbD02147 does not bind to human AKT2 and AKT3 (81% and 82% identical to AKT1, respectively).

Two serine/threonine kinases, designated AKT1 and AKT2 exhibit sequence homology with the protein kinase A and C families and are encoded by the c-AKT proto-oncogene. AKT1 and the related AKT2 are activated by platelet-derived growth factor (PDGF). The activation is rapid and specific, and it is abrogated by mutations in the Pleckstrin homology domain (PH) of AKT1. It was shown that the activation occurs through phosphatidylinositol 3-kinase. Binding of the PH domain to the phosphatidylinositol 3-kinase alpha (PI(3)K) results in its targeting to the plasma membrane. Additionally, the activation of AKT1 and AKT2 is inhibited by the PI kinase inhibitor wortmannin. Taken together, this data strongly suggests that the protein signals downstream of the PI kinases. In the developing nervous system AKT is a critical mediator of growth factor-induced neuronal survival. Survival factors can suppress apoptosis in a transcription-independent manner by activating the serine/threonine kinase AKT1, which then phosphorylates and inactivates components of the apoptotic machinery.
Phosphorylation on Thr-308, Ser-473 and Tyr-474 is required for full activity. Ser-473 is dephosphorylated by PH domain leucine-rich repeat protein phosphatase (Gao et al. 2005).

Product Details

Target Species
Human
Species Cross-Reactivity
Target SpeciesCross Reactivity
Rat Expected from Sequence
Mouse Expected from Sequence
N.B. Antibody reactivity and working conditions may vary between species.
Product Form
A lyophilised bivalent human recombinant Fab selected from the HuCAL® GOLD phage display library. Expressed in E. coli and purified using NiNTA affinity chromatography. This Fab fragment is dimerized via a helix-turn-helix motif. The antibody is tagged with a myc-tag (EQKLISEEDL) and a his-tag (HHHHHH) at the C-terminus of the antibody heavy chain.
Reconstitution
Reconstitute with 100 μl of distilled water.
Preparation
Metal-chelate affinity chromatography.
Buffer Solution
Phosphate buffered saline
Preservative Stabilisers
None present.
Immunogen
Human AKT1 (Val-106 - Ala-480) fusion protein. The antigen contains the complete kinase domain (amino acids 150 to 408) but not the N-terminal PH domain. Amino acid residues 106-480, MW 77.8 kDa.
Activity
Activity was tested by indirect ELISA: recombinant purified GST-AKT1 (5 μg/ml) plus unrelated control proteins were immobilized on a microtiter plate. Specific binding was monitored by first adding HCA003 (2 μg/ml), then adding a secondary antibody (goat anti-human F(ab')2 fragment specific, AP conjugate. 1:5000 diluted). A fluorescent signal was created by adding the AP substrate Attophos. The signal on the antigen is at least 5-fold above background, whereas the signal on the control antigens is less than 1.5-fold above background.
Purity
Purity was tested by SDS-PAGE and Coomassie-staining of a 2 μg sample.
Approx. Protein Concentrations
Antibody concentration 1.0 mg/ml following reconstitution.

Storage Information

Storage
Prior to reconstitution store at +4oC.
After reconstitution store at -20oC.
Storage in frost-free freezers is not recommended. Avoid repeated freezing and thawing as this may denature the antibody.
Shelf Life
6 months from date of reconstitution.

More Information

UniProt
P31749 Related reagents
Entrez Gene
AKT1 Related reagents
GO Terms
GO:0005886 plasma membrane
GO:0000060 protein import into nucleus, translocation
GO:0005829 cytosol
GO:0004674 protein serine/threonine kinase activity
GO:0005524 ATP binding
GO:0005547 phosphatidylinositol-3,4,5-trisphosphate binding
GO:0005654 nucleoplasm
GO:0005978 glycogen biosynthetic process
GO:0006417 regulation of translation
GO:0006469 negative regulation of protein kinase activity
GO:0006809 nitric oxide biosynthetic process
GO:0006924 activation-induced cell death of T cells
GO:0007186 G-protein coupled receptor protein signaling pathway
GO:0008286 insulin receptor signaling pathway
GO:0008629 induction of apoptosis by intracellular signals
GO:0008633 activation of pro-apoptotic gene products
GO:0042802 identical protein binding
GO:0009408 response to heat
GO:0010748 negative regulation of plasma membrane long-chain fatty acid transport
GO:0010975 regulation of neuron projection development
GO:0016071 mRNA metabolic process
GO:0046326 positive regulation of glucose import
GO:0046889 positive regulation of lipid biosynthetic process
GO:0019899 enzyme binding
GO:0018105 peptidyl-serine phosphorylation
GO:0048015 phosphatidylinositol-mediated signaling
GO:0030168 platelet activation
GO:0030235 nitric-oxide synthase regulator activity
GO:0030307 positive regulation of cell growth
GO:0031018 endocrine pancreas development
GO:0031295 T cell costimulation
GO:0031659 positive regulation of cyclin-dependent protein kinase activity involved in G1/S
GO:0031999 negative regulation of fatty acid beta-oxidation
GO:0034405 response to fluid shear stress
GO:0043325 phosphatidylinositol-3,4-bisphosphate binding
GO:0045429 positive regulation of nitric oxide biosynthetic process
GO:0045600 positive regulation of fat cell differentiation
GO:0045725 positive regulation of glycogen biosynthetic process
GO:0046777 protein autophosphorylation
GO:0048009 insulin-like growth factor receptor signaling pathway
GO:0048011 nerve growth factor receptor signaling pathway
GO:0051000 positive regulation of nitric-oxide synthase activity
GO:0070141 response to UV-A
GO:0090004 positive regulation of establishment of protein localization in plasma membrane
Acknowledgements
Sold under license of U.S. Patents 6753136, 7785859 and 8273688 and corresponding patents. This antibody was developed by Bio-Rad, Zeppelinstr. 4, 82178 Puchheim, Germany.
His-tag is a registered trademark of EMD Biosciences.
Western blotting data kindly provided by ProQinase GmbH, Freiburg.
Licensed Use
For in vitro research purposes only, unless otherwise specified in writing by Bio-Rad.
Regulatory
For research purposes only

Applications of AKT1 antibody

This product has been reported to work in the following applications. This information is derived from testing within our laboratories, peer-reviewed publications or personal communications from the originators. Please refer to references indicated for further information. For general protocol recommendations, please visit the antibody protocols page.
Application Name Verified Min Dilution Max Dilution
ELISA 2ug/ml
Western Blotting 2ug/ml
Where this product has not been tested for use in a particular technique this does not necessarily exclude its use in such procedures. Suggested working dilutions are given as a guide only. It is recommended that the user titrates the product for use in their own system using appropriate negative/positive controls.
Technical Advice
Recommended protocols and further information about HuCAL recombinant antibody technology can be found in the HuCAL Antibodies Technical Manual

Secondary Antibodies Available

Description Product Code Pack Size Applications List Price Quantity
Goat anti Human IgG F(ab')2:HRP 0500-0099 0.5 ml E WB
Mouse anti c-Myc MCA2200 1 mg C E F* P WB*
Mouse anti Penta Histidine Tag:HRP MCA5995P 125 µl E WB