Anticorps Recombinant de lapin anti-Phospho-AKT (Thr308)

Phospho-AKT (Thr308) Recombinant Antibody for WB, FC (Intra), ELISA

Hôte / Isotype

Lapin / IgG

Réactivité testée

Humain, rat, souris

Applications

WB, FC (Intra), ELISA

Conjugaison

Non conjugué

CloneNo.

242063B2

N° de cat : 81232-10-RR

Synonymes

AKT (Thr308), p-AKT, p-AKT (Thr308), p-AKT Thr308, p-AKT-Thr308

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Galerie de données de validation

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  WB analysis using 81232-10-RR

  Non-treated HEK-293 cells and Calyculin A treated HEK-293 cells were subjected to SDS PAGE followed by western blot with 81232-10-RR (Phospho-AKT (Thr308) antibody) at dilution of 1:5000 incubated at room temperature for 1.5 hours. The membrane was stripped and re-blotted with Beta Actin (66009-1-Ig) antibody as a loading control.

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  WB analysis using 81232-10-RR

  Non-treated NIH/3T3 cells and Calyculin A treated NIH/3T3 cells were subjected to SDS PAGE followed by western blot with 81232-10-RR (Phospho-AKT (Thr308) antibody) at dilution of 1:5000 incubated at room temperature for 1.5 hours. The membrane was stripped and re-blotted with Beta Actin (66009-1-Ig) antibody as a loading control.

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  WB analysis using 81232-10-RR

  Non-treated HSC-T6 cells and Calyculin A treated HSC-T6 cells were subjected to SDS PAGE followed by western blot with 81232-10-RR (Phospho-AKT (Thr308) antibody) at dilution of 1:5000 incubated at room temperature for 1.5 hours. The membrane was stripped and re-blotted with Beta Actin (66009-1-Ig) antibody as a loading control.

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  FC experiment of HeLa using 81232-10-RR

  1X10^6 HeLa cells untreated (dashed lines) or treated with Calyculin A which intracellularly stained with 0.13 ug Phospho-AKT (Thr308) Recombinant antibody (81232-10-RR, Clone:242063B2) and CoraLite®488-Conjugated Goat Anti-Rabbit IgG(H+L) (SA00013-2)(red), or 0.13 ug Rabbit IgG Isotype Control RecAb (98136-1-RR, Clone: 240953C9) (blue). Cells were fixed with 4% PFA and permeabilized with 90% MeOH.

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Applications testées

• Résultats positifs en WB: cellules HSC-T6 traitées à la calyculine A, cellules HEK-293 traitées à la calyculine A, cellules NIH/3T3 traitées à la calyculine A
• Résultats positifs en FC (Intra): cellules HeLa traitées à la calyculine A,

Dilution recommandée

• Western Blot (WB): WB : 1:2000-1:10000
• Flow Cytometry (FC) (INTRA): FC (INTRA) : 0.13 ug per 10^6 cells in a 100 µl suspension

It is recommended that this reagent should be titrated in each testing system to obtain optimal results.

Sample-dependent, check data in validation data gallery

Informations sur le produit

81232-10-RR cible Phospho-AKT (Thr308) dans les applications de WB, FC (Intra), ELISA et montre une réactivité avec des échantillons Humain, rat, souris

• Réactivité: Humain, rat, souris
• Hôte / Isotype: Lapin / IgG
• Clonalité: Recombinant
• Type: Anticorps
• Immunogène: Peptide
• Nom complet: v-akt murine thymoma viral oncogene homolog 1
• Masse moléculaire calculée: 56 kDa
• Poids moléculaire observé: 60 kDa
• Numéro d’acquisition GenBank: BC000479
• Symbole du gène: AKT1
• Identification du gène (NCBI): 207
• Conjugaison: Non conjugué
• Forme: Liquide
• Méthode de purification: Purification par protéine A
• Tampon de stockage: PBS with 0.02% sodium azide and 50% glycerol
• Conditions de stockage: Stocker à -20°C. Stable pendant un an après l'expédition. L'aliquotage n'est pas nécessaire pour le stockage à -20^oC Les 20ul contiennent 0,1% de BSA.

Informations générales

AKT is a serine/threonine kinase and it participates in the key role of the PI3K signaling pathway. Phosphatidylinositol-3 kinase (PI3K) is the key regulator of AKT activation. The recruitment of inactive AKT protein to PIP3-rich areas of the plasma membrane results in a conformational change that exposes the activation loop of AKT. AKT's activating kinase, phosphoinositide-dependent protein kinase (PDK1) , is also recruited to PIP3 microdomains. PDK1 phosphorylates AKT on threonine 308 (Thr308) of the exposed activation loop, activating AKT and leading to a second phosphorylation of AKT at serine 473 (Ser473) by a kinase presumed to be mTORC2 that further potentiates kinase activity. Active AKT will phosphorylate various downstream protein targets that control cell growth and translational control and act to suppress apoptosis. (PMID: 31594388, PMID: 30808672)

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