Human MAP4K5 Baculovirus-Insect cells Overexpression Lysate

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Human MAP4K5 Baculovirus-Insect cells Overexpression Lysate: Product Information

Product Description
This Human MAP4K5 overexpression lysate was created in Baculovirus-Insect cells and intented for use as a Western blot (WB) positive control. Purification of MAP4K5 protein (Cat: 11773-H20B) from the overexpression lysate was verified.
Expression Host
Baculovirus-Insect cells
Species
Human
Sequence Information
A DNA sequence encoding the human MAP4K5 (NP_006566.2) (Met 1-Tyr 846) was fused with the N-terminal polyhistidine-tagged GST tag at the N-terminus.
Molecule Mass
The recombinant human MAP4K5/GST chimera consists of 1083 amino acids and has a calculated molecular mass of 123 kDa. It migrates as an approximately 116 kDa band in SDS-PAGE under reducing conditions.

Human MAP4K5 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 MAP4K5 Baculovirus-Insect cells Overexpression Lysate: Alternative Names

Human GCKR Overexpression Lysate; Human KHS Overexpression Lysate; Human KHS1 Overexpression Lysate; Human MAPKKKK5 Overexpression Lysate

MAP4K5 Background Information

Mitogen-activated protein kinase kinase kinase kinase 5, also known as Kinase homologous to SPS1/STE2, MAPK/ERK kinase kinase kinase 5, MEK kinase kinase 5 and MAP4K5, is a cytoplasm protein which belongs to theprotein kinase superfamily, STE Ser/Thr protein kinase family and STE2 subfamily. MAP4K5 is ubiquitously expressed in all tissues examined, with high levels in the ovary, testis and prostate. It contains oneCNH domain and oneprotein kinase domain. MAP4K5 is highly similar to yeast SPS1/STE2 kinase. Yeast SPS1/STE2 functions near the beginning of the MAP kinase signal cascades that is essential for yeast pheromone response. MAP4K5 has been shown to interact with CRKL and TRAF2. This kinase was shown to activate Jun kinase in mammalian cells. MAP4K5 is an early component of MAP kinase signal cascades. It may play a role in the response to environmental stress. MAP4K5 appears to act upstream of the JUN N-terminal pathway.
Full Name
mitogen-activated protein kinase kinase kinase kinase 5
References
  • Schultz SJ, et al.,1994, Cell Growth Differ. 4 (10): 821-30.
  • Tung RM, et al.,1997, Oncogene 14 (6): 653-9.
  • Shi, C S, et al., 1999, J. Immunol.163 (6): 3279-85.
  • Shi, C S, et al., 2000, Blood. 95 (3): 776-82. 
  • Gu,Y. et al., 2006, J Hum Genet  51 (7): 605-10. 
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