Human TLK2 Baculovirus-Insect cells Overexpression Lysate: Product Information
This Human TLK2 overexpression lysate was created in Baculovirus-Insect cells and intented for use as a Western blot (WB) positive control. Purification of TLK2 protein (Cat: 11551-H20B) from the overexpression lysate was verified.
A DNA sequence encoding the amino acids (Leu 397-Asn 772) of human TLK2 (Q86UE8-1) was fused with the N-terminal polyhistidine-tagged GST tag at the N-terminus.
The recombinant human TLK2 (aa 397-772)/GST chimera consists of 613 amino acids and has a calculated molecular mass of 71.3 kDa. It migrates as an approximately 65 kDa band in SDS-PAGE under reducing conditions.
Human TLK2 Baculovirus-Insect cells Overexpression Lysate: Usage Guide
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.
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.
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.
Western Blot (WB) Optimal dilutions/concentrations should be determined by the end user.
Human TLK2 Baculovirus-Insect cells Overexpression Lysate: Alternative Names
Human HsHPK Overexpression Lysate; Human PKU-ALPHA Overexpression Lysate
TLK2 Background Information
Serine / threonine-protein kinase tousled-like 2, also known as PKU-alpha, Tousled-like kinase 2 and TLK2, is a nucleus protein which belongs to theprotein kinase superfamily and Ser/Thr protein kinase family. The tousled-like kinases are an evolutionarily conserved family of proteins implicated in DNA repair, DNA replication and mitosis in metazoans and plants. Their absence from the yeasts and other eukaryotic 'microbes' suggests a specific role for them in the development of multicellular organisms. Tousled-like kinase 2 / TLK2 is widely expressed. It is present in fetal placenta, liver, kidney, pancreas, heart and skeletal muscle. It is also found in adult cell lines. Tousled-like kinase 2 / TLK2 contains oneprotein kinase domain. Tousled-like kinase 2 / TLK2 is rapidly and transiently inhibited by phosphorylation following the generation of DNA double-stranded breaks during S-phase. This is cell cycle checkpoint and ATM-pathway dependent and appears to regulate processes involved in chromatin assembly.
tousled-like kinase 2
Sillje HHW. et al., 2001,Curr Biol. 11: 1068-73.
Groth A. et al., 2003, EMBO J. 22: 1676-87.
Beausoleil SA. et al., 2004, Proc Natl Acad Sci. 101: 12130-5.
Mayya V. et al., 2009, Sci Signal. 2: RA46.
Oppermann FS. et al., 2009, Mol Cell Proteomics. 8: 1751-64.
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