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Human Cbl-c / CBL-3 Insect Cell Lysate (WB positive control)

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Human CBLC Transfected / Overexpression Cell Lysate Product Information
Expressed Host:Baculovirus-Insect cells
Product Description:Baculovirus-Insect Cell lysate that Human CBLC / Cbl-c transfected / overexpressed for Western blot (WB) positive control. The whole cell lysate is provided in 1X Sample Buffer (1X modified RIPA buffer+1X SDS loading buffer).
Sequence information:A DNA sequence encoding the human CBLC (AAH28915.1) (Met1-Ala474) was expressed with the N-terminal polyhistidine-tagged GST tag at the N-terminus.
Predicted N Terminal:Met
Molecule Mass:The recombinant human CBLC chimera consists of 711 amino acids and has a calculated molecular mass of 80.2 kDa. The recombinant protein migrates approximately 66 kDa band in SDS-PAGE under reducing conditions.
Human CBLC Transfected / Overexpression Cell Lysate Usage Guide
Preparation Method:Cell lysate was prepared by homogenization in ice-cold modified RIPA Lysis Buffer with cocktail of protease inhibitors (Sigma). Cell debris was removed by centrifugation. Protein concentration was determined by Bradford assay (Bio-Rad protein assay, Microplate Standard assay). The cell lysate was boiled for 5 min in 1 x SDS loading buffer (50 mM Tris-HCl pH 6.8, 12.5% glycerol, 1% sodium dodecylsulfate, 0.01% bromophenol blue) containing 5% b-mercaptoethanol, and lyophilized.
Lysis Buffer:Modified RIPA Lysis Buffer: 50 mM Tris-HCl pH 7.4, 150 mM NaCl, 1mM EDTA, 1% Triton X-100, 0.1% SDS, 1% Sodium deoxycholate, 1mM PMSF.
Quality Control Testing:12.5% SDS-PAGE Stained with Coomassie Blue after protein purification.
Stability:Samples are stable for up to twelve months from date of receipt.
Recommend Usage:1.  Centrifuge the tube for a few seconds and ensure the pellet at the bottom of the tube. 2.  Re-dissolve the pellet using 200μL pure water and boil for 2-5 min. 3. Store the lyophilized cell lysate at 4℃. After re-dissolution, recommend to aliquot it into smaller quantities and store at -80℃.
Storage Buffer:1 X Sample Buffer (1 X modified RIPA buffer+1 X SDS loading buffer).
Storage Instruction:Store at 4℃. After re-dissolution, aliquot and store at -80℃.
Application notes:Western blot (WB): Use at an assay dependent dilution.
Other Applications: Not tested.
Optimal dilutions/concentrations should be determined by the end user.
Cbl-c / CBL-3 Background

CBL proteins, such as Cbl-c, are phosphorylated upon activation of a variety of receptors that signal via protein tyrosine kinases. Through interactions with proteins containing SRC homology-2 (SH2) and SH3 domains, CBL proteins modulate downstream cell signaling. Cbl-c is a member of the Cbl family of E3 ubiquitin ligases. Expression of Cbl-c gene may be restricted to epithelial cells, and alternatively spliced transcript variants encoding multiple isoforms have been observed for Cbl-c gene.

Human Cbl-c / CBL-3 References
  • Kim M. et al., 1999, Gene. 239 (1): 145-54.
  • Courbard JR. et al., 2003, J. Biol. Chem. 277 (47): 45267-75.
  • Keane MM. et al., 1999, Oncogene. 18 (22): 3365-75.
  • Size / Price
    Catalog: 15659-H20BL-300
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