44-1230G

antibody from Invitrogen Antibodies

Targeting: FOXO3 AF6q21, FKHRL1, FOXO2, FOXO3A

Antibody data

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Validation data

FOXO3 protein structure - 44-1230G shown in red.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Lysates prepared from Hek293 cells transfected with wild type GFP-FOXO3 alone mutant S207A GFP-FOXO3 co-transfected with wild type MST1, or wild-type FOXO3 co-transfected with wild-type active MST1, were resolved on a 10% polyacrylamide gel and transferred to PVDF. The membrane was blocked with a 3% BSA-TBST buffer for one hour at room temperature, and then incubated with the FOXO3 [pS207] antibody for two hours at room temperature in 3% BSA-TBST buffer, following prior incubation with: no peptide, the non-phosphopeptide corresponding to the phosphopeptide immunogen, a generic phosphoserine containing peptide, or the phosphopeptide immunogen. After washing, the membrane was incubated with goat F(ab')2 anti-rabbit IgG HRP conjugate (Prod #ALI4404) and signals were detected using the Pierce SuperSignal™ method. The data show that MST1 induced the phosphorylation of FOXO3 at serine 207 and that the signal was abolished in cells over-expressing mutant S207A FOXO3 or in the presence of inactive MST1 kinase. The data also show that the signal is blocked only by the phosphopeptide corresponding to FOXO3 [pS207], indicating again its specificity.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Lysates prepared from Hek293 cells transfected with wild type GFP-FOXO3 alone mutant S207A GFP-FOXO3 co-transfected with wild type MST1, or wild-type FOXO3 co-transfected with wild-type active MST1, were resolved on a 10% polyacrylamide gel and transferred to PVDF. The membrane was blocked with a 3% BSA-TBST buffer for one hour at room temperature, and then incubated with the FOXO3 [pS207] antibody for two hours at room temperature in 3% BSA-TBST buffer, following prior incubation with: no peptide, the non-phosphopeptide corresponding to the phosphopeptide immunogen, a generic phosphoserine containing peptide, or the phosphopeptide immunogen. After washing, the membrane was incubated with goat F(ab')2 anti-rabbit IgG HRP conjugate (Prod #ALI4404) and signals were detected using the Pierce SuperSignal™ method. The data show that MST1 induced the phosphorylation of FOXO3 at serine 207 and that the signal was abolished in cells over-expressing mutant S207A FOXO3 or in the presence of inactive MST1 kinase. The data also show that the signal is blocked only by the phosphopeptide corresponding to FOXO3 [pS207], indicating again its specificity.