Human GFR Alpha-2/GFRA2 HEK293 Overexpression Lysate: 产品信息
This Human GFR Alpha-2/GFRA2 overexpression lysate was created in HEK293 Cells and intented for use as a Western blot (WB) positive control. Purification of GFR Alpha-2/GFRA2 protein (Cat: 10331-H08H) from the overexpression lysate was verified.
A DNA sequence encoding the human GFRα2 (NP_001486.4) without the propeptide (Met 1-Ser 441) was fused with a polyhistidine tag at the C-terminus.
The recombinant human GFRα2 consists of 431 amino acids after removal of the signal peptide and predicts a molecular mass of 48.2 kDa. As a result of glycosylation, the apparent molecular mass of rhGFRα2 is approximately 60-70 kDa in SDS-PAGE under reducing conditions.
Human GFR Alpha-2/GFRA2 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 GFR Alpha-2/GFRA2 HEK293 Overexpression Lysate: 别称
Human GDNFRB Overexpression Lysate; Human NRTNR-ALPHA Overexpression Lysate; Human NTNRA Overexpression Lysate; Human RETL2 Overexpression Lysate; Human TRNR2 Overexpression Lysate
GFR Alpha-2/GFRA2 背景信息
GFRA2 is a member of the GDNF receptor family. It is a glycosylphosphatidylinositol(GPI)-linked cell surface receptor for both GDNF and NTN, and mediates activation of the RET tyrosine kinase receptor. GFRA2 is a potent survival factor for central and peripheral neurons, and is essential for the development of kidneys and the enteric nervous system. Glial cell line-derived neurotrophic factor (GDNF) and neurturin (NTN) are its binding ligand which are two structurally related, potent neurotrophic factors that play key roles in the control of neuron survival and differentiation. GDNF promotes the formation of a physical complex between GFRA/GDNFRa and the orphan tyrosin kinase receptor Ret, thereby inducing its tyrosine phosphorylation. The RET is a receptor tyrosine kinase representing the signal-transducing molecule of a multisubunit surface receptor complex for the GDNF, in which GFRA/GDNFRa acts as the ligand-binding component. Experiments have improved that GFRA2 genetic variants and age may play a role in Tardive dyskinesia (TD) susceptibility, but further work is required to confirm these findings.
GDNF family receptor alpha 2
Jing S, et al. (1997) GFRalpha-2 and GFRalpha-3 are two new receptors for ligands of the GDNF family. J Biol Chem. 272(52): 33111-7.
Souza RP, et al. (2010) Glial cell line-derived neurotrophic factor receptor alpha 2 (GFRA2) gene is associated with tardive dyskinesia. Psychopharmacology. 210(3): 347-54.
Vanhorne JB, et al. (2001) Cloning and characterization of the human GFRA2 locus and investigation of the gene in Hirschsprung disease. Hum Genet. 108(5): 409-15.