This Human MFAP5 overexpression lysate was created in HEK293 Cells and intented for use as a Western blot (WB) positive control. Purification of MFAP5 protein (Cat: 12648-H08H) from the overexpression lysate was verified.
A DNA sequence encoding the human MFAP5 (Q13361) (Met1-Leu173) with a C-terminal polyhistidine tag was expressed.
The recombinant human MFAP5 comprises 163 amino acids and has a predicted molecular mass of 18.7 kDa. The apparent molecular mass of the protein is approximately 54 and 29 kDa in SDS-PAGE under reducing conditions.
Human MFAP5 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 MFAP5 HEK293 Overexpression Lysate: 别称
Human AAT9 Overexpression Lysate; Human MAGP-2 Overexpression Lysate; Human MAGP2 Overexpression Lysate; Human MFAP-5 Overexpression Lysate; Human MP25 Overexpression Lysate
MFAP5 (Microfibril Associated Protein 5, also known as MAGP2) is a Protein Coding gene. MFAP5 is a component of the elastin-associated microfibrils. It belongs to the MFAP family. As a 25-kD microfibril-associated glycoprotein, MFAP5 is rich in serine and threonine residues. It stimulates the assembly of elastic fibers, a complex structure composed of a tropoelastin inner core and microfibril outer mantle comprising proteins such as fibrillins and microfibril-associated glycoproteins that guide tropoelastin deposition. MFAP5 also stabilizes type 1 procollagen and thus plays an important role in extracellular matrix homeostasis. It has multiple binding regions on fibrillins and has a covalent periodic association with fibrillin-containing microfibrils. Diseases associated with MFAP5 include Aortic Aneurysm, Familial Thoracic 9, and Familial Thoracic Aortic Aneurysm And Aortic Dissection.
microfibrillar associated protein 5
Hatzinikolas G, et al. (1998) The exon structure of the human MAGP-2 gene. Similarity with the MAGP-1 gene is confined to two exons encoding a cysteine-rich region. J Biol Chem. 273(45):29309-14.
Gibson MA, et al. (1996) Further characterization of proteins associated with elastic fiber microfibrils including the molecular cloning of MAGP-2 (MP25). J Biol Chem. 271(2): 1096-103.
Miyamoto A, et al. (2006) Microfibrillar proteins MAGP-1 and MAGP-2 induce Notch1 extracellular domain dissociation and receptor activation. J Biol Chem. 281(15):10089-97.