This Cynomolgus CD59 overexpression lysate was created in HEK293 Cells and intented for use as a Western blot (WB) positive control. Purification of CD59 protein (Cat: 90188-C08H) from the overexpression lysate was verified.
A DNA sequence encoding the cynomolgus CD59 (G7PQF7) (Met1-Glu101) was expressed with a polyhistidine tag at the C-terminus.
The recombinant cynomolgus CD59 comprises 87 amino acids and has a calculated molecular mass of 10.2 KDa. The apparent molecular mass of it is approximately 18 and 14 KDa respectively in SDS-PAGE.
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.
Cynomolgus CD59 HEK293 Overexpression Lysate: 别称
Cynomolgus CD59 Overexpression Lysate
CD59 glycoprotein, also known as 2 kDa homologous restriction factor, HRF2, MAC-inhibitory protein, Membrane attack complex inhibition factor, Membrane inhibitor of reactive lysis, MIC11, MIRL and CD59, is a cell membrane protein which contains one UPAR/Ly6 domain. CD59 is a small, highly glycosylated, GPI-linked protein, with a wide expression profile. The soluble form of CD59 from urine retains its specific complement binding activity, but exhibits greatly reduced ability to inhibit MAC assembly on cell membranes. CD59 is a potent inhibitor of the complement membrane attack complex (MAC) action. CD59 was first identified as a regulator of the terminal pathway of complement. It acts by binding to the C8 and/or C9 complements of the assembling MAC, thereby preventing incorporation of the multiple copies of C9 required for complete formation of the osmolytic pore. This inhibitor appears to be species-specific. CD59 is involved in signal transduction for T-cell activation complexed to a protein tyrosine kinase. Defects in CD59 are the cause of CD59 deficiency (CD59D).
Complement Activation Pathways
Fletcher CM. et al., 1994, Structure. 2: 185-99.
Rudd PM. et al., 1997, J Biol Chem. 272: 7229-44.
Kimberley FC. et al., 2007, Mol Immunol. 44 (1-3): 73-81.
Gong Y. et al., 2007, Sci China C Life Sci. 50 (6): 773-9.
Picariello G. et al., 2008, Proteomics 8: 3833-47.
Heibeck TH. et al., 2009, J Proteome Res. 8: 3852-61.