This Human RAB27B overexpression lysate was created in HEK293 Cells and intented for use as a Western blot (WB) positive control. Purification of RAB27B protein (Cat: 16007-H04H) from the overexpression lysate was verified.
A DNA sequence encoding the human RAB27B (O00194) (Thr2-Lys215) was expressed with the Fc region of mouse IgG1 at the N-terminus.
The recombinant human RAB27B/mFc comprises 450 amino acids and has a predicted molecular mass of 50.8 kDa. The apparent molecular mass of the monomer is approximately 51 kDa in SDS-PAGE under reducing conditions due to glycosylation.
Human RAB27B 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 RAB27B HEK293 Overexpression Lysate: 别称
Human C25KG Overexpression Lysate; Human RAB27B Overexpression Lysate
RAB27B is a member of the Rab family GTPases involved in vesicle trafficking. The Rab27 subfamily consists of Rab27a and Rab27b. The human RAB27B gene is organized in six exons, spanning about 69 kb in the chromosome 18q21.1 region. The Rab27 subfamily of Ras-like GTPases is highly conserved in mammals. There is high degree of conservation in sequence and gene structure between RAB27A and RAB27B genes. Exogenous expression of Rab27b in melanocytes results in melanosomal association as observed for Rab27a, suggesting the two Rab27 proteins are functional homologues. As with RAB27A in Griscelli Disease, RAB27B may be also associated with human disease mapping to chromosome 18.