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9794
c-Cbl Antibody Sampler Kit
Primary Antibodies
Antibody Sampler Kit

c-Cbl Antibody Sampler Kit #9794

Citations (0)
Western blot analysis of cell extracts from various cell types using c-Cbl (C49H8) Rabbit mAb.
Western blot analysis of extracts from 293-T cells transfected with c-Cbl and Fyn (lanes 3 and 6) or without Fyn (lanes 2 and 5), untreated or calf intestinal phosphatase (CIP)-treated, using Phospho-c-Cbl (Tyr731) Antibody. (Lysates provided by Dr. A.L. Reddi, laboratory of Dr. Hamid Band, Harvard University, Massachusetts
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 Jurkat cells, untreated (-) or treated with pervanadate (1 mM; +), using Phospho-c-Cbl (Tyr700) (D16D7) Rabbit mAb (upper) or c-Cbl (D4E10) Rabbit mAb #8447 (lower).
Western blot analysis of extracts from Jurkat cells, untreated (-) or treated with pervanadate (1 mM; +), using Phospho-c-Cbl (Tyr700) (D16D7) Rabbit mAb (left). The phospho-specificity of the antibody was verified by preincubating the antibody with c-Cbl (Tyr700) phosphopeptide (center) or with c-Cbl (Tyr700) nonphosphopeptide prior to incubation with the membrane (right).
Immunoprecipitation (IP) of phosphorylated c-Cbl from Jurkat cell extracts, untreated (-) or treated with pervanadate (1 mM; +), using Phospho-c-Cbl (Tyr700) (D16D7) Rabbit mAb (lanes 4 and 6). Western blot detection was performed using the same antibody. Lanes 1 and 2 are 10% input. Lanes 3 and 5 are IPs performed with Rabbit (DA1E) mAb IgG XP® Isotype Control #3900.
Inquiry Info.# 9794

Product Description

The c-Cbl Antibody Sampler Kit provides an economical means of evaluating levels of c-Cbl protein phosphorylated at the specified sites, as well as total c-Cbl levels. The kit contains enough reagents to perform four western blot experiments per primary antibody.

Specificity / Sensitivity

The Phospho-c-Cbl (Tyr700) and (Tyr731) Antibodies detect endogenous c-Cbl when phosphorylated at the specified sites. c-Cbl (C49H3) Rabbit mAb detects endogenous levels of total c-Cbl protein.

Source / Purification

The phospho-specific polyclonal antibody is produced by immunizing animals with a synthetic phosphopeptide corresponding to residues surrounding Tyr731 of human c-Cbl. Polyclonal antibodies are purified by protein A and peptide affinity chromatography. The monoclonal antibodies are produced by immunizing animals with synthetic peptides corresponding to the sequence of human c-Cbl and residues surrounding Tyr700 of human c-Cbl.

Background

The c-Cbl proto-oncogene is a ubiquitously expressed cytoplasmic adaptor protein that is especially predominant in hematopoietic cells (1,2). c-Cbl is rapidly tyrosine-phosphorylated in response to stimulation of a variety of cell-surface receptors and becomes associated with a number of intracellular signaling molecules such as protein tyrosine kinases, phosphatidylinositol-3 kinase, Crk, and 14-3-3 proteins (3,4). c-Cbl possesses a highly conserved amino-terminal phosphotyrosine binding domain (TKB) and a C3HC4 RING finger motif. The TKB recognizes phosphorylated tyrosines on activated receptor tyrosine kinases (RTKs) as well as other nonreceptor tyrosine kinases. The RING finger motif recruits ubiquitin-conjugating enzymes. These two domains are primarily responsible for the ubiquitin ligase activity of c-Cbl and downregulation of RTKs (3). Research studies have indicated that in human cancer tissues, c-Cbl is frequently tyrosine-phosphorylated in a tumor-specific manner (5). Phosphorylation of Tyr731 of c-Cbl provides a docking site for downstream signaling components such as p85 and Fyn (6).

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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U.S. Patent No. 7,429,487, foreign equivalents, and child patents deriving therefrom.
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