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XML generation date: 2024-09-20 06:21:54.596
Product last modified at: 2024-10-02T11:00:09.917Z
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PDP - Template Name: Antibody Sampler Kit
PDP - Template ID: *******4a3ef3a

Forkhead Signaling Antibody Sampler Kit #9946

    Product Information

    Product Description

    This sampler kit provides an economical means to investigate Forkhead signaling. The kit contains primary and secondary antibodies to perform two Western blots with each antibody.

    Specificity / Sensitivity

    Phospho-FoxO1 (Thr24)/FoxO3a (Thr32) Antibody detects endogenous levels of FoxO1/FoxO3a only when phosphorylated at Thr24 of FoxO1 or Thr32 of FoxO3a. The antibody cross-reacts with phosphorylated FoxO4 at Thr28, but not with FoxO1 family members phosphorylated at other sites. Phospho-FoxO1 (Ser256) Antibody detects endogenous levels of FoxO1 only when phosphorylated at Ser256. The antibody cross-reacts with Fox04 phosphorylated at Ser193. Phospho-FoxO1 (Thr24)/FoxO3a (Thr32)/Fox04 (Thr28) (4G6) Rabbit mAb detects endogenous levels of FoxO1 when phosphorylated at Thr24, of FoxO3a when phosphorylated at Thr32 or FoxO4 when phosphorylated at Thr28. FoxO1 (C29H4) Rabbit mAb detects endogenous levels of total FoxO1 protein. The antibody does not detect exogenously expressed family members FoxO3a or FoxO4. Phospho-FoxO3a (Ser253) (D18H8) Rabbit mAb detects endogenous levels of FoxO3a only when phosphorylated at Ser253. This antibody may cross-react with FoxO1 when overexpressed and phosphorylated at Ser251 or FoxO4 when overexpressed and phosphorylated at Ser197. Phospho-FoxO3a (Ser318/321) Antibody detects endogenous levels of FoxO3a only when phosphorylated at Ser318/321. The antibody is expected to cross-react with FoxO1 when phosphorylated at Ser322/325 based on the peptide sequence. FoxO3a (D19A7) Rabbit mAb recognizes endogenous levels of total FoxO3a protein. The FoxO4 Antibody detects endogenous levels of FoxO4. The antibody is sensitive to phosphorylation within the antigen and preferrentially detects unphosphorylated FoxO4.

    Source / Purification

    Polyclonal antibodies are produced by immunizing animals with a synthetic phosphopeptide corresponding to residues surrounding Thr28 of human Fox04, with a synthetic phosphopeptide corresponding to residues around Ser256 of human Fox01, or with a synthetic peptide corresponding to residues of human Fox04. Polyclonal antibodies are purified by protein A and peptide affinity chromatography. Monoclonal antibodies are produced by immunizing animals with recombinant protein specific to the carboxy terminus of human FoxO3 protein, with a GST-fusion protein corresponding to carboxy-terminal residues of human FoxO1, with a synthetic phosphopeptide corresponding to the sequence of human Fox03a, or with a synthetic phosphopeptide corresponding to residues surrounding Thr28 of human Fox04.

    Background

    The Forkhead family of transcription factors is involved in tumorigenesis of rhabdomyosarcoma and acute leukemias (1-3). Within the family, three members (FoxO1, FoxO4, and FoxO3a) have sequence similarity to the nematode orthologue DAF-16, which mediates signaling via a pathway involving IGFR1, PI3K, and Akt (4-6). Active forkhead members act as tumor suppressors by promoting cell cycle arrest and apoptosis. Increased expression of any FoxO member results in the activation of the cell cycle inhibitor p27 Kip1. Forkhead transcription factors also play a part in TGF-β-mediated upregulation of p21 Cip1, a process negatively regulated through PI3K (7). Increased proliferation results when forkhead transcription factors are inactivated through phosphorylation by Akt at Thr24, Ser256, and Ser319, which results in nuclear export and inhibition of transcription factor activity (8). Forkhead transcription factors can also be inhibited by the deacetylase sirtuin (SirT1) (9).
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