R1467

antibody from OriGene

Targeting: AKT2 PKBβ

Antibody data

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Validation data

AKT2 protein structure - R1467 shown in red.

Supportive validation

Submitted by: OriGene (provider)
Main image
Experimental details: Figure 1. Immunoblotting. Affinity Purified antibody to AKT2 was used at a 1:1,000 dilution to detect AKT2 by Western blot. A lysate from a stable HEK293 cell line expressing an inducible, myristoylated form of Akt2 (MyrAkt2-ER) was loaded for SDSPAGE, separated and then transferred to nitrocellulose. The blot was reacted with primary antibody for 1h at room temperature. In response to 4OHT (tamoxifen), the Akt2 is recruited to the plasma membrane via its myristoylation sequence and becomes phosphorylated and activated. The blot (right panel) shows 0 m, 15 m, and 30 m of 4OHT treatment. This treatment has no effect on endogenous Akt2, but causes a band shift upwards in the MyrAkt2. Endogenous Akt2 runs at about 60kDa whereas the myristoylated construct runs at around 110 kDa.
Validation comment: WB

Supportive validation

Submitted by: OriGene (provider)
Main image
Experimental details: Figure 2. Immunoblotting. Affinity Purified antibody to AKT2 was used at a 1:1,000 dilution to detect AKT2 by Western blot in lysates from mouse tissues. The antibody is shown to react with mouse Akt2 in mouse liver, skeletal muscle and fat using standard immunoblotting methods.
Validation comment: WB

Supportive validation

Submitted by: OriGene (provider)
Main image
Experimental details: Figure 3. Immunoblotting. Antibody to AKT2 detects both unphosphorylated and phosphorylated AKT2. As before approximately 20 mg/lane of crude HEK293 lysate was loaded for SDS-PAGE, separated and then transferred to nitrocellulose. The right lane contains lysate pretreated for 15 min with 25 uM LY294002 which affects the phosphorylation of endogenous Akt2 but has no effect on phosphorylation of the myristoylated construct. This antibody clearly detects both the phosphorylated (top arrow) and the non-phosphorylated (bottom arrow) forms of endogenous Akt2.
Validation comment: WB