Human Germinal Center Kinase/MAP4K2 Baculovirus-Insect cells Overexpression Lysate

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Human Germinal Center Kinase/MAP4K2 Baculovirus-Insect cells Overexpression Lysate: Product Information

Product Description
This Human Germinal Center Kinase/MAP4K2 overexpression lysate was created in Baculovirus-Insect cells and intented for use as a Western blot (WB) positive control. Purification of Germinal Center Kinase/MAP4K2 protein (Cat: 11826-H20B) from the overexpression lysate was verified.
Expression Host
Baculovirus-Insect cells
Species
Human
Sequence Information
A DNA sequence encoding the human MAP4K2 (AAH47865.1) (Met 1-Tyr 812) was fused with the N-terminal polyhistidine-tagged GST tag at the N-terminus.
Molecule Mass
The recombinant human MAP4K2/GST chimera consists of 1049 amino acids and has a calculated molecular mass of 119 kDa. It migrates as an approximately 116 kDa band in SDS-PAGE under reducing conditions.

Human Germinal Center Kinase/MAP4K2 Baculovirus-Insect cells 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 Germinal Center Kinase/MAP4K2 Baculovirus-Insect cells Overexpression Lysate: Alternative Names

Human BL44 Overexpression Lysate; Human GCK Overexpression Lysate; Human MAP4K2 Overexpression Lysate; Human RAB8IP Overexpression Lysate

Germinal Center Kinase/MAP4K2 Background Information

Mitogen-activated protein kinase kinase kinase kinase 2, also known as B lymphocyte serine/threonine-protein kinase, Germinal center kinase, MAPK/ERK kinase kinase kinase 2, MEK kinase kinase 2, Rab8-interacting protein and MAP4K2, is a cytoplasm and peripheral membrane protein which belongs to theprotein kinase superfamily, STE Ser/Thr protein kinase family and STE2 subfamily. MAP4K2 contains oneCNH domain and oneprotein kinase domain. Although this kinase is found in many tissues, its expression in lymphoid follicles is restricted to the cells of germinal centre, where it may participate in B-cell differentiation. MAP4K2 can be activated by TNF-alpha, and has been shown to specifically activate MAP kinases. It is also found to interact with TNF receptor-associated factor 2 (TRAF2), which is involved in the activation of MAP3K1 / MEKK1. MAP4K2 enhances MAP3K1 oligomerization, which may relieve amino-terminal mediated MAP3K1 autoinhibition and lead to activation following autophosphorylation. It may also play a role in the regulation of vesicle targeting or fusion.
Full Name
mitogen-activated protein kinase kinase kinase kinase 2
References
  • Katz P, et al.,1994, J Biol Chem 269 (24): 16802-9.
  • Ren, M; et al., 1996,  Proc. Natl. Acad. Sci. USA. 93 (10): 5151-5.
  • Chadee D.N., et al., 2002, Mol. Cell. Biol. 22:737-49.
  • Wissing J., et al., 2007, Mol. Cell. Proteomics 6:537-47.
  • Oppermann F.S., et al., 2009, Mol. Cell. Proteomics 8:1751-64.
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