Human MST4 Baculovirus-Insect cells Overexpression Lysate

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Human MST4 Baculovirus-Insect cells Overexpression Lysate: Product Information

Product Description
This Human MST4 overexpression lysate was created in Baculovirus-Insect cells and intented for use as a Western blot (WB) positive control. Purification of MST4 protein (Cat: 10667-H09B) from the overexpression lysate was verified.
Expression Host
Baculovirus-Insect cells
Species
Human
Sequence Information
A DNA sequence encoding the human MST4 isoform 1 (NP_057626.2) (Met 1-Pro 416) was expressed with the fused GST tag at N-terminus.
Molecule Mass
The recombinant human MST41/GST chimera consists of 641 amino acids and predicts a molecular mass of 73 kDa. It migrates as an approximately 65 kDa band in SDS-PAGE under reducing conditions.

Human MST4 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 MST4 Baculovirus-Insect cells Overexpression Lysate: Alternative Names

Human MASK Overexpression Lysate; Human MST4 Overexpression Lysate

MST4 Background Information

MST4, also known as mammalian STE2-like protein kinase 4, is a novel member of the germinal center kinase subfamily of human Ste2-like kinases and is closely related to MST3. The 416 amino acid full-length MST4 contains a C-terminal regulatory domain and an N-terminal kinase domain, both of which are required for full activation of the kinase. MST4 is highly expressed in placenta, thymus, and peripheral blood leukocytes. MST4 specifically activates ERK but not JNK or p38 MAPK in transient transfected cells or in stable cell lines, and thus is biologically active in the activation of MEK/ERK pathway mediating cell growth and transformation. Further, MST4 kinase activity is stimulated significantly by epidermal growth factor receptor (EGFR) ligands, which are known to promote growth of certain cancer cells. Accordingly, MST4 have a potential role in signal transduction pathways involved in cancer progression. Three alternatively spliced isoform of MST4 have been isolated, and isoform 3 lacks an exon encoding kinase domain and may function as a dominant-negative regulator of the MST4 kinase.
Full Name
serine/threonine protein kinase 26
References

    1. Qian, Z. et al., 2001, J Biol Chem. 276 :22439-45.

    2. Lin, JL. et al., 2001, Oncogene. 20: 6559-6569.

    3. Sung V, et al., 2003, Cancer research. 63: 3356-63.

    4. Ma, X. et al., 2007, Molecular biology of the cell. 18:1965-78.

    5. ten Klooster JP, et al., 2009, Developmental cell. 16:551-62.

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