This Human JTB overexpression lysate was created in HEK293 Cells and intented for use as a Western blot (WB) positive control. Purification of JTB protein (Cat: 13447-H05H) from the overexpression lysate was verified.
A DNA sequence encoding the human JTB (O76095-1) (Met1-Leu105) was fused with Fc region of mouse IgG at the C-terminus.
The recombinant human JTB/mFc is a disulfide-linked homodimer. The reduced monomer comprises 309 amino acids and has a predicted molecular mass of 34.7 kDa. The apparent molecular mass of the protein is approximately 38 in SDS-PAGE under reducing conditions due to glycosylation.
Human JTB HEK293 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.
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 JTB HEK293 Overexpression Lysate: 别称
Human hJT Overexpression Lysate; Human HJTB Overexpression Lysate; Human HSPC222 Overexpression Lysate; Human JTB Overexpression Lysate; Human PAR Overexpression Lysate
Jumping translocation breakpoint, also known as JTB, is a member of the JTB family. Jumping translocation (JT) is an unbalanced translocation that comprises amplified chromosomal segments jumping to various telomeres. JTB is expressed in all normal human tissues studied but overexpressed or underexpressed in many of their malignant counterparts. It is required for normal cytokinesis during mitosis. JTB plays a role in the regulation of cell proliferation. It may be a component of the chromosomal passenger complex (CPC), a complex that acts as a key regulator of mitosis. The CPC complex has essential functions at the centromere in ensuring correct chromosome alignment and segregation and is required for chromatin-induced microtubule stabilization and spindle assembly.
jumping translocation breakpoint
Hatakeyama S. et al., 1999, Oncogene. 18 (12): 2085-90.
Platica O. et al., 2000, Int J Oncol. 16 (5): 1055-61.
Erhard DS. et al., 2004, Genome Res. 14 (10B): 2121-7.