Human Hemojuvelin Baculovirus-Insect cells Overexpression Lysate: 产品信息
This Human Hemojuvelin overexpression lysate was created in Baculovirus-Insect cells and intented for use as a Western blot (WB) positive control. Purification of Hemojuvelin protein (Cat: 10410-H08B) from the overexpression lysate was verified.
A DNA sequence encoding the mature form of human HFE2 isoform a (Q6ZVN8-1) (Met 1-Ser 399) was fused with a polyhistidine tag at the C-terminus.
The secreted recombinant human HFE2 consists of 374 amino acids and predicts a molecular mass of 40 kDa. As a result of intracellular cleavage, the apparent molecular mass of the protein is approximately 20, 34 and 44 KDa in SDS-PAGE under reducing conditions, corresponding to the N-terminal, C-terminal portions and the full-length respectively.
Human Hemojuvelin 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.
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 Hemojuvelin Baculovirus-Insect cells Overexpression Lysate: 别称
Human HFE2A Overexpression Lysate; Human HJV Overexpression Lysate; Human JH Overexpression Lysate; Human RGMC Overexpression Lysate
Hemojuvelin, also known as HFE2, is a membrane-bound and soluble protein that belongs to the repulsive guidance molecule (RGM) family. It is known that RGMs function through Neogenin, a homolog of the Netrin receptor deleted in colon cancer. In mammals, the RGM family consists of three glycoproteins that have discrete expression patterns and functions (RGM-A, RGM-B, and RGM-C). Hemojuvelin is expressed in adult and fetal liver, heart, and skeletal muscle. Hemojuvelin acts as a bone morphogenetic protein (BMP) coreceptor. Enhancement of BMP signaling regulates hepcidin (HAMP) expression and iron metabolism. It plays a key role in iron metabolism. Hemojuvelin represents the cellular receptor for hepcidin. It may be a component of the signaling pathway that activates hepcidin or it may act as a modulator of hepcidin expression. Defects in hemojuvelin are the cause of hemochromatosis type 2A, also known as juvenile hemochromatosis (JH).
hemochromatosis type 2 (juvenile)
Papanikolaou G, et al. (2004) Mutations in HFE2 cause iron overload in chromosome 1q-linked juvenile hemochromatosis. Nat Genet. 36(1):77-82.
Babitt JL, et al. (2006) Bone morphogenetic protein signaling by hemojuvelin regulates hepcidin expression. Nat Genet. 38(5):531-9.
Zhang AS, et al. (2008) Neogenin-mediated hemojuvelin shedding occurs after hemojuvelin traffics to the plasma membrane. J Biol Chem. 283(25):17494-502.