Human DAPK1 Baculovirus-Insect cells Overexpression Lysate

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Human DAPK1 Baculovirus-Insect cells Overexpression Lysate: Product Information

Product Description
This Human DAPK1 overexpression lysate was created in Baculovirus-Insect cells and intented for use as a Western blot (WB) positive control. Purification of DAPK1 protein (Cat: 11966-H20B) from the overexpression lysate was verified.
Expression Host
Baculovirus-Insect cells
Species
Human
Sequence Information
A DNA sequence encoding the N-terminal segment of human DAPK1 (P53355-1) (Met 1-Leu 363) was fused with the N-terminal polyhistidine-tagged GST tag at the N-terminus.
Molecule Mass
The recombinant human DAPK1 (aa 1-363)/GST chimera consists of 600 amino acids and has a calculated molecular mass of 69.4 kDa. It migrates as an approximately 64 kDa band in SDS-PAGE under reducing conditions.

Human DAPK1 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 DAPK1 Baculovirus-Insect cells Overexpression Lysate: Alternative Names

Human DAPK Overexpression Lysate; Human ROCO3 Overexpression Lysate

DAPK1 Background Information

Death-associated protein kinase 1, also known as DAP kinase 1, DAPK1 and DAPK, is a cytoplasm protein which belongs to theprotein kinase superfamily, CAMK Ser / Thr protein kinase family and DAP kinase subfamily. DAPK1 contains tenANK repeats, onedeath domain and oneprotein kinase domain. DAPK1 is a calcium / calmodulin-dependent serine/threonine kinase which acts as a positive regulator of apoptosis. DAPK1 gene is a candidate tumor suppressor (TSG) and the abnormal methylation of DAPK1 gene has been found in many carcinomas. DAPK1 over-expression can induce cell apoptosis and inhibit tumor cell metastasis. DAPK1 gene over-expression could suppress PGCl3 cells malignant phenotype, inhibit PGCl3 cells growth, invasive, migration and adhesion ability, upregulate p53 gene and downregulate bcl-2 gene. Loss of activity of death-associated protein kinase 1 ( DAPK1 ) may be an independent factor affecting survival of non-small cell lung cancer patients. DAPK1 promoter methylation might play a significant role in the progression of chronic myeloid leukemia ( CML ).
Full Name
death-associated protein kinase 1
References
  • Zhang,H.T. et al., 2004, Ai Zheng. 23 (5):497-501.
  • Martoriati,A. et al., 2005, Oncogene. 24 (8):1461-6.
  • Li,Y. et al., 2006, Hum Mol Genet. 15 (17): 2560-8.
  • Martinez-Glez,V. et al., 2007, Neoplasma. 54 (2):123-6.
  • Raval,A. et al., 2007, Cell. 129 (5):879-90.
  • Gade,P. et al., 2009, Int J Cancer. 125 (7):1566-74.
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