Human TRIB2 Baculovirus-Insect Overexpression Lysate 产品信息
Product Description
This Human TRIB2 overexpression lysate was created in Baculovirus-Insect Cells and intented for use as a Western blot (WB) positive control. Purification of TRIB2 protein (Cat: 10725-H19B) from the overexpression lysate was verified.
Expression Host
Baculovirus-Insect Cells
Sequence Information
A DNA sequence encoding the human TRIB2 (NP_067675.1) (Met 1-Asn 343) was fused with the GST tag at the N-terminus and a polyhistidine tag at the C-terminus.
Molecule Mass
The recombinant human TRIB2/GST chimera consists of 578 amino acids and predicts a molecular mass of 66 kDa.
Human TRIB2 Baculovirus-Insect Overexpression Lysate Usage Guide
Preparation Method
Cell lysate was prepared by homogenization of the over-expressed cells 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.
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.
Sample Buffer
1 X Sample Buffer (1 X modified RIPA buffer+1 X SDS loading buffer).
Stability & Storage
Store at 4℃ for up to twelve months from date of receipt. After re-dissolution, aliquot and store at -80℃ for up to twelve months. Avoid repeated freeze-thaw cycles.
Application
Western Blot (WB)
Optimal dilutions/concentrations should be determined by the end user.
Human TRIB2 Baculovirus-Insect Overexpression Lysate Alternative Names
Human C5FW Overexpression Lysate;Human FLJ57420 Overexpression Lysate;Human GS3955 Overexpression Lysate;Human TRB2 Overexpression Lysate;Human TRIB2 Overexpression Lysate
TRIB2 Background Information
Tribbles homolog 2, also known as TRB-2, and Trib2, is a member of the protein kinase superfamily and Tribbles subfamily (Trib1, Trib2, Trib3). The identification of tribbles as regulators of signal processing systems and physiological processes, including development, together with their potential involvement in diabetes and cancer, has generated considerable interest in these proteins. Tribbles have been reported to regulate activation of a number of intracellular signalling pathways with roles extending from mitosis and cell activation to apoptosis and modulation of gene expression. Tribbles controls the timing of mitosis in the prospective mesoderm, allowing cell-shape changes to be completed. This mechanism for coordinating cell division and cell-shape changes may have helped Drosophila to evolve its mode of rapid early development. Trib2 was identified as a downregulated transcript in leukemic cells undergoing growth arrest. Trib2-transduced bone marrow cells exhibited a growth advantage and readily established factor-dependent cell lines. Trib2-reconstituted mice uniformly developed fatal transplantable acute myelogenous leukemia (AML).
Full Name
tribbles pseudokinase 2
References
Seher, TC. et al., 2000, Curr Biol. 10 (11): 623-9. Keeshan, K. et al., 2006, Cancer Cell. 10 (5): 401-11. Hegedus, Z. et al., 2007, Cell Signal. 19 (2):238-50. Keeshan, K. et al., 2008, Blood Cells Mol Dis. 40 (1): 119-21. Cvetkovic, LV. et al., 2010, J Clin Invest. 120 (3): 713-9.
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