This Human RPRD1B overexpression lysate was created in HEK293 Cells and intented for use as a Western blot (WB) positive control. Purification of RPRD1B protein (Cat: 14027-H07H) from the overexpression lysate was verified.
A DNA sequence encoding the human RPRD1B (AAH33629.1) (Met1-Asp326) was expressed with an N-terminal polyhistidine tag.
The recombinant human RPRD1B comprises 346 amino acids and has a predicted molecular mass of 39.3 kDa. The apparent molecular mass of the protein is approximately 39 kDa in SDS-PAGE under reducing conditions due to glycosylation.
Human RPRD1B 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 RPRD1B HEK293 Overexpression Lysate: 别称
Human C20orf77 Overexpression Lysate; Human CREPT Overexpression Lysate; Human dJ1057B20.2 Overexpression Lysate; Human NET60 Overexpression Lysate
RPRD1B, together with RPRD1A, can accompany RNAP II from promoter regions to 3'-untranslated regions during transcription in vivo, predominantly interact with phosphorylated RNAP II, and can reduce CTD S5- and S7-phosphorylated RNAP II at target gene promoters. RNA polymerase II C-terminal domain (CTD) phosphorylation is important for various transcription-related processes. RPRD1B is a transcriptional regulator that enhances expression of CCND1. It also enhances the transcription of a number of other cell cycle-related genes including CDK2, CDK4, CDK6 and cyclin-E but not CDKN1A, CDKN1B or cyclin-A.
regulation of nuclear pre-mRNA domain containing 1B
Ni Z. et al., 2011, Transcription. 2 (5): 237-42.
Kristensen AR. et al., 2012, Nat Methods. 9 (9): 907-9.