Human VLDL Receptor HEK293 Overexpression Lysate Product Information
This Human VLDL Receptor overexpression lysate was created in HEK293 Cells and intented for use as a Western blot (WB) positive control. Purification of VLDL Receptor protein (Cat: 11075-H08H) from the overexpression lysate was verified.
A DNA sequence encoding the human VLDLR isoform alpha (NP_003374.3) extracellular domain (Met 1-Ser 797) was fused with a polyhistidine tag at the C-terminus.
The recombinant human VLDLR comprises 781 amino acids with a predicted molecular mass of 86 kDa. It migrates with the apparent molecular weight of 150 & 180 kDa due to different glycosylation in SDS-PAGE under reducing conditions.
Human VLDL Receptor HEK293 Overexpression Lysate Usage Guide
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.
Modified RIPA Lysis Buffer: 50 mM Tris-HCl pH 7.4, 150 mM NaCl, 1mM EDTA, 1% Triton X-100, 0.1% SDS, 1% Sodium deoxycholate, 1mM PMSF.
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).
Stability & Storage
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 VLDL Receptor HEK293 Overexpression Lysate Alternative Names
Human CAMRQ1 Overexpression Lysate;Human CARMQ1 Overexpression Lysate;Human CHRMQ1 Overexpression Lysate;Human VLDLRCH Overexpression Lysate
VLDL Receptor Background Information
The very low density lipoprotein receptor, known as VLDLR, is a single-pass type 1 integral membrance protein and a member of the LDL receptor family. This receptor family includes LDL receptor, LRP, megalin, VLDLR and ApoER2, and is characterized by a cluster of cysteine-rich class A repeats, epidermal growth factor (EGF)-like repeats, YWTD repeats and an O-linked sugar domain. VLDLR contains 3 EGF-like domains, 8 LDL-receptor class A domains, as well as 6 LDL-receptor class B repeats, and is abundant in heart, skeletal muscle, also ovary and kidney, but not in liver. VLDLR binds VLDL and transports it into cells by endocytosis. In order to be internalized, the receptor-ligand complexes must first cluster into clathrin-coated pits. VLDLR mediates the phosphorylation of mDab1 (mammalian disabled protein) via binding to Reelin, and induces the modulation of Tau phosphorylation. This pathway regulates the migration of neurons along the radial glial fiber network during brain development. Defects of VLDLR may be the cause of VLDLR-associated cerebellar hypoplasia (VLDLRCH), a syndrome characterized by moderate-to-profound mental retardation, delayed ambulation, and predominantly truncal ataxia.
very low density lipoprotein receptor
Trommsdorff, M. et al., 1999. Cell. 97: 689-701. Mikhailenko, I. et al., 1999. J. Cell Sci. 112: 3269-3281. Sato, A. et al., 1999. Biochem. J. 341: 377-383. Hiesberger, T. et al., 1999. Neuron 24: 481-489. Tiebel, O. et al., 1999. Atherosclerosis 145: 239-251. Boycott, K.M. et al., 2005, Am. J. Hum. Genet. 77 (3): 477-483. Moheb, L.A. et al., 2008, Eur. J. Hum. Genet. 16 (2): 270-273.