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05-669

Sigma-Aldrich

Anti-phospho-Akt1/PKBα (Ser473) Antibody, clone 11E6

clone 11E6, Upstate®, from mouse

Synonym(s):

Protein kinase B, RAC-alpha serine/threonine-protein kinase, murine thymoma viral (v-akt) oncogene homolog-1, rac protein kinase alpha, v-akt murine thymoma viral oncogene homolog 1

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About This Item

UNSPSC Code:
12352203
eCl@ss:
32160702
NACRES:
NA.41

biological source

mouse

Quality Level

antibody form

purified antibody

antibody product type

primary antibodies

clone

11E6, monoclonal

species reactivity

rat, canine, mouse, human

manufacturer/tradename

Upstate®

technique(s)

ELISA: suitable
western blot: suitable

isotype

IgG1κ

NCBI accession no.

UniProt accession no.

shipped in

dry ice

target post-translational modification

phosphorylation (pSer473)

Gene Information

dog ... Akt1(490878)
human ... AKT1(207)

General description

The serine/threonine kinase Akt family contains several members, including Akt1 (also designated PKB or RacPK), Akt2 (also designated PKB-β or RacPK-β) and Akt3 (also designated PKB-γ or thyoma viral proto-oncogene 3), which exhibit sequence homology with the protein kinase A and C families and are encoded by the c-Akt proto-oncogene. All members of the Akt family have a Pleckstrin homology domain. Akt1 and Akt2 are activated by PDGF stimulation. This activation is dependent on PDGFR-β tyrosine residues 740 and 751, which bind the 85 kDa subunit of the phosphatidylinositol 3-kinase (PI 3-kinase) complex. Activation of Akt1 by insulin or insulin-growth factor-1(IGF-1) results in phosphorylation of both Thr 308 and Ser 473. Phosphorylation of both residues is important to generate a high level of Akt1 activity, and the phosphorylation of Thr 308 is not dependent on phosphorylation of Ser 473 in vivo. Thus, Akt proteins become phosphorylated and activated in insulin/IGF-1- stimulated cells by an upstream kinase(s). 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.

Specificity

Recognizes phosphorylated Akt1/PKBα at Ser473, Mr 60 kDa. This lot of antibody does not cross-react with Akt1/PKBα dephosphorylated at Ser473.

Application

ELISA:
A previous lot was recommended at 0.05 mg/mL by an independent laboratory for ELISA.
This Anti-Akt1 Antibody, clone 11E6 is validated for use in ELISA, WB for the detection of phospho-Akt1/PKBα (Ser473).

Quality

Evaluated by western blot on lysates from mouse NIH-3T3 fibroblasts treated with PDGF.

Western Blot Analysis: 0.5-2 µg/mL of this antibody detected phosphorylated Akt1/PKBa in lysates from mouse NIH-3T3 fibroblasts treated with 100 ng/mL PDGF for 20 minutes.

Target description

60 kDa

Physical form

Format: Purified
Purified mouse monoclonal IgG1κ lyophilized from 1 mL 2X PBS, PEG, sucrose, and 0.09% sodium azide.
Reconstitute with 1 mL H2O (15 min. at RT). Concentration after reconstitution: 100 µg/mL

Storage and Stability

Stable for 1 year at -20°C from date of receipt.
Reconstitute prior to use. Reconstitute with 1 mL H2O for 15 min at room temperature. Reconstituted aliquots should be stored frozen at -80°C. Thawed aliquots may be stored at 2-8°C for up to 3 months. Avoid repeated freeze/thaw cycles, which may damage IgG and affect product performance.

Analysis Note

Control
PDGF treated NIH-3T3 cell lysates.

Other Notes

Concentration: Please refer to the Certificate of Analysis for the lot-specific concentration.

Legal Information

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

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Pictograms

Skull and crossbones

Signal Word

Danger

Hazard Statements

Hazard Classifications

Acute Tox. 3 Dermal - Acute Tox. 4 Inhalation - Acute Tox. 4 Oral - Aquatic Chronic 3

Storage Class Code

6.1C - Combustible acute toxic Cat.3 / toxic compounds or compounds which causing chronic effects

WGK

WGK 1


Certificates of Analysis (COA)

Search for Certificates of Analysis (COA) by entering the products Lot/Batch Number. Lot and Batch Numbers can be found on a product’s label following the words ‘Lot’ or ‘Batch’.

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