Human TLK1 Baculovirus-Insect cells Overexpression Lysate


Human TLK1 Baculovirus-Insect cells Overexpression Lysate: 产品信息

This Human TLK1 overexpression lysate was created in Baculovirus-Insect cells and intented for use as a Western blot (WB) positive control. Purification of TLK1 protein (Cat: 12213-H20B) from the overexpression lysate was verified.
Baculovirus-Insect cells
A DNA sequence encoding the human TLK1 isoform 1 (Q9UKI8-1) (Met 1-Tyr 766) was fused with the N-terminal polyhistidine-tagged GST tag at the N-terminus.
The recombinant human TLK1/GST chimera consists of 1003 amino acids and has a calculated molecular mass of 114 kDa. It migrates as an approximately 125 kDa band in SDS-PAGE under reducing conditions.

Human TLK1 Baculovirus-Insect cells Overexpression Lysate: 使用指南

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.
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.
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).
稳定性 & 储存条件
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 TLK1 Baculovirus-Insect cells Overexpression Lysate: 别称

Human KIAA0137 Overexpression Lysate; Human PKU-beta Overexpression Lysate

TLK1 背景信息

Tousled-like kinase 1 (or protein kinase ubiquitous, PKU-beta/TLK1) is a serine/threonine protein kinase that is implicated in chromatin remodeling, DNA replication and mitosis. TLK1 has a function important for proper chromosome segregation and maintenance of diploid cells at mitosis in mammalian cells that could be mediated by reduced phosphorylation of histone H3 and condensation of chromosomes, although other explanations to the phenotype are possible.
tousled like kinase 1
  • Sillje H.H.W., et al., 2001, Curr. Biol. 11:1068-73.
  • Beausoleil S.A., et al., 2004, Proc. Natl. Acad. Sci. USA. 101:12130-5.
  • Oppermann F.S., et al., 2009, Mol. Cell. Proteomics 8:1751-64.
  • Mayya V., et al., 2009, Sci. Signal. 2: RA46-RA46.
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