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9652
PTEN and PDK1 Antibody Sampler Kit
Primary Antibodies
Antibody Sampler Kit

PTEN and PDK1 Antibody Sampler Kit #9652

Citations (7)
Simple Western™ analysis of lysates (1 mg/mL) from HeLa cells using PTEN (D4.3) XP® Rabbit mAb #9188. The virtual lane view (left) shows the target band (as indicated) at 1:10 and 1:50 dilutions of primary antibody. The corresponding electropherogram view (right) plots chemiluminescence by molecular weight along the capillary at 1:10 (blue line) and 1:50 (green line) dilutions of primary antibody. This experiment was performed under reducing conditions on the Jess™ ​​​​​​​ Simple Western instrument from ProteinSimple, a BioTechne brand, using the 12-230 kDa separation module.
Western blot analysis of extracts from SW-13, NIH/3T3, Jurkat and PC12 cells, using PDK1 Antibody.
Western blot analysis of extracts from PC3 cells, HCT116 wild-type and HCT116 PDK1 -/- cells using Phospho-PDK1 (Ser241) (C49H2) Rabbit mAb (upper) and Akt (pan) (C67E7) Rabbit mAb #4691 (lower). (HCT116 wild-type and HCT116 PDK1 -/- cells were kindly provided by Dr. Bert Vogelstein, Johns Hopkins University, Baltimore, MD).
After the primary antibody is bound to the target protein, a complex with HRP-linked secondary antibody is formed. The LumiGLO® is added and emits light during enzyme catalyzed decomposition.
Western blot analysis of extracts from various cell lines using Non-phospho PTEN (Ser380/Thr382/Thr383) (D2D11) Rabbit mAb. The non-phosphospecificity of the antibody was verified by preincubating the antibody without peptide (-), with PTEN (Ser380/Thr382/Thr383) non-phosphopeptide (+), or with PTEN (Ser380/Thr382/Thr383) phosphopeptide (+) prior to incubating the membrane.
Western blot analysis of extracts from various cell lines using PTEN (D4.3) XP® Rabbit mAb (upper) and Akt (pan) (C67E7) Rabbit mAb #4691 (lower).
Immunohistochemical analysis of paraffin-embedded human esophageal adenocarcinoma using PTEN (D4.3) XP® Rabbit mAb performed on the Leica BOND RX.
Western blot analysis of extracts from HeLa and NIH/3T3 cells, using Phospho-PTEN (Ser380/Thr382/383) (44A7) Rabbit mAb. Membranes were either left untreated (-) or treated with (+) calf intestinal phosphatase (CIP) post Western transfer to verify phospho-specificity of the antibody.
Western blot analysis of extracts from PC3 cells, untreated or λ phosphatase-treated, using Phospho-PDK1 (Ser241) (C49H2) Rabbit mAb (upper), PDK1 Antibody #3062 (middle) or Akt Antibody #9272 (lower).
Immunohistochemical analysis of paraffin-embedded human prostate adenocarcinoma using PTEN (D4.3) XP® Rabbit mAb performed on the Leica BOND RX.
Immunohistochemical analysis of paraffin-embedded human squamous cell lung carcinoma using PTEN (D4.3) XP® Rabbit mAb performed on the Leica BOND RX.
Immunohistochemical analysis of paraffin-embedded human colon using PTEN (D4.3) XP® Rabbit mAb.
Immunohistochemical analysis using PTEN (D4.3) XP® Rabbit mAb on SignalSlide(TM) PTEN IHC Controls #8106 (paraffin-embedded LNCaP (left) and NIH/3T3 (right) cells).
Inquiry Info.# 9652

Product Description

The PTEN and PDK1 Sampler Kit provides an economical means to evaluate two key enzymes that regulate multiple signaling pathways. The kit contains enough primary and secondary antibodies to perform two Western blots per primary antibody.

Specificity / Sensitivity

Each antibody in the PTEN and PDK1 Antibody Sampler Kit detects endogenous levels of its target protein. Activation state antibodies detect only target proteins phosphorylated at indicated residues. Non-Phospho-PTEN (Ser380/Thr382/Thr383) Antibody detects endogenous levels of PTEN only when dephosphorylated at Ser380, Thr382 and Thr383.

Source / Purification

Phospho-specific rabbit monoclonal antibodies are produced by immunizing animals with synthetic phosphopeptides corresponding to residues around Ser380, Thr382 and Thr383 of human PTEN and around Ser241 of human PDK1. PTEN (D4.3) XP® Monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to residues in the carboxy-terminal sequence of human PTEN. Monoclonal antibody Non-phospho PTEN (Ser380/Thr382/Thr383) (D2D11) is produced by immunizing animals with a synthetic non-phosphopeptide corresponding to residues surrounding Ser380/Thr382/Thr383 of human PTEN protein. Polyclonal PDK1 antibody is produced by immunizing animals with synthetic peptides corresponding to residues surrounding the carboxy terminus of human PDK1. Polyclonal antibodies are purified using protein A and peptide affinity chromatography.

Background

PTEN (phosphatase and tensin homologue deleted on chromosome ten), also referred to as MMAC (mutated in multiple advanced cancers) phosphatase, is a tumor suppressor implicated in a wide variety of human cancers (1). PTEN encodes a 403 amino acid polypeptide originally described as a dual-specificity protein phosphatase (2). The main substrates of PTEN are inositol phospholipids generated by the activation of the phosphoinositide 3-kinase (PI3K) (3). PTEN is a major negative regulator of the PI3K/Akt signaling pathway (1,4,5). PTEN possesses a carboxy-terminal, noncatalytic regulatory domain with three phosphorylation sites (Ser380, Thr382, and Thr383) that regulate PTEN stability and may affect its biological activity (6,7). PTEN regulates p53 protein levels and activity (8) and is involved in G protein-coupled signaling during chemotaxis (9,10).
Phosphoinositide-dependent protein kinase 1 (PDK1) plays a central role in many signal transduction pathways (11,12) including the activation of Akt and the PKC isoenzymes p70 S6 kinase and RSK (13). Through its effects on these kinases, PDK1 is involved in the regulation of a wide variety of processes, including cell proliferation, differentiation and apoptosis.

Pathways

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Limited Uses

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For Research Use Only. Not for Use in Diagnostic Procedures.
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