This Human Fetuin B overexpression lysate was created in HEK293 Cells and intented for use as a Western blot (WB) positive control. Purification of Fetuin B protein (Cat: 11834-H08H) from the overexpression lysate was verified.
A DNA sequence encoding the human FETUB (NP_055190.2) (Met 1-Pro 382) was expressed, with a polyhistidine tag at the C-terminus.
The recombinant human FETUB consists of 375 amino acids and predictes a molecular mass of 41.7 kDa. rhFETUB migrates as an approximately 55 kDa band in SDS-PAGE under reducing conditions due to glycosylation.
Human Fetuin B 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 Fetuin B HEK293 Overexpression Lysate: 别称
Human 16G2 Overexpression Lysate; Human Gugu Overexpression Lysate; Human IRL685 Overexpression Lysate
Fetuin B 背景信息
Fetuin-B, also known as Fetuin-like protein IRL685 and FETUB, is a secreted protein which belongs to thefetuin family. Fetuin-B / FETUB contains twocystatin domains. Fetuin-B is a member of the fetuin family, part of the cystatin superfamily of cysteine protease inhibitors. Fetuins have been implicated in several diverse functions, including osteogenesis and bone resorption. Fetuin-A has been identified as a major protein during fetal life and is also involved in important functions such as protease inhibitory activities and development-associated regulation of calcium metabolism and osteogenesis. Fetuin-A is a key partner in the recovery phase of an acute inflammatory response. Fetuin-B / FETUB is found at least in human and rodents. It is unambiguously a paralogue of Fetuin-A. Fetuin-A and Fetuin-B exhibit significant differences at the amino acid sequence level, notably including variations with respect to the archetypal fetuin-specific signature.
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