A simple and rapid anti-ligand enzyme immunoassay for visualization of low-density lipoprotein membrane receptors.
Triton X-114 was used to solubilize the membrane proteins of bovine adrenal cortex and human leukocytes. The solubilized membrane proteins were subjected to electrophoresis and transblotted to nitrocellulose paper and incubated with LDL/acetyl-LDL. The combination of peroxidase-conjugated second antibody and 4-chloronaphthol/H2O2 allowed rapid development of colored bands where LDL or acetyl-LDL bound to electroblotted proteins. The ELISA is highly sensitive and efficient for screening a large number of samples and avoids the need for a continuous supply of radiolabeled antibodies and autoradiography.
D000302:Adrenal Cortex / Q000032:analysis; D000818:Animals; D002417:Cattle; D002462:Cell Membrane / Q000032:analysis; D004591:Electrophoresis, Polyacrylamide Gel; D004797:Enzyme-Linked Immunosorbent Assay; D006801:Humans; D007962:Leukocytes / Q000032:analysis; D008079:Lipoproteins, VLDL / Q000032:analysis*; D008565:Membrane Proteins / Q000302:isolation & purification; D008970:Molecular Weight; D011973:Receptors, LDL / Q000032:analysis* / Q000378:metabolism; D012995:Solubility
D008079:Lipoproteins, VLDL; D008565:Membrane Proteins; D011973:Receptors, LDL; C036053:VLDL receptor