Human LDLR Quantikine ELISA Kit Summary

Product Summary

The Quantikine Human LDL R Immunoassay is a 4.5 hour solid-phase ELISA designed to measure human LDL R in cell culture supernates, cell lysates, serum, heparin plasma, and urine. It contains NS0-expressed recombinant human LDL R and has been shown to accurately quantitate the recombinant factor. Results obtained using natural human LDL R showed linear curves that were parallel to the standard curves obtained using the Quantikine kit standards. These results indicate that this kit can be used to determine relative mass values for naturally occurring human LDL R.

Preparation and Storage

Background: LDLR

LDL R (Low Density Lipoprotein Receptor) is a widely expressed cell surface scavenger receptor. LDL R binds ApoB and ApoE, the apolipoproteins of low- and very low-density lipoproteins (LDL and VLDL), respectively. Hepatocyte LDL R is responsible for endocytosis and clearing of most plasma LDL cholesterol. At the low pH of the endocytic vesicle, it dissociates, allowing degradation of LDL and recycling of LDL R to the cell surface. Lack of LDL R expression or function causes familial hypercholesterolemia (FH). The protease PCSK9 can also cause increased plasma cholesterol by promoting LDL R degradation rather than recycling to the cell surface.

Assay Procedure

Refer to the product datasheet for the complete assay procedure.

Bring all reagents and samples to room temperature before use. It is recommended that all samples, standards, and controls be assayed in duplicate.

1、Prepare all reagents, standard dilutions, and samples as directed in the product insert.

2、Remove excess microplate strips from the plate frame, return them to the foil pouch containing the desiccant pack, and reseal.

3、Add 50 μL of Assay Diluent to each well.

4、Add 50 μL of Standard, control, or sample to each well. Cover with a plate sealer, and incubate at room temperature for 2 hours.

5、Aspirate each well and wash, repeating the process 4 times for a total of 5 washes.

6、Add 100 μL of Conjugate to each well. Cover with a new plate sealer, and incubate at room temperature for 2 hours.

7、Aspirate and wash 5 times.

8、Add 100 μL Substrate Solution to each well.

9、Add 100 μL of Stop Solution to each well. Read at 450 nm within 30 minutes. Set wavelength correction to 540 nm or 570 nm.