To determine whether the antibodies raised were indeed specific against the target protein, ELISA against intact, full-length target proteins (PCKS9 or LDL) was performed. First, 96-well PolySorp plates [Thermo Fisher Scientific] were coated using 100 μl of recombinant mouse PCKS9 [Abcam] (10 μg/ml in PBS) or 100μl of human LDL [Thermo Fisher Scientific] (10μg/ml in PBS) or 100μl human oxidized LDL [Thermo Fisher Scientific] (10μg/ml in PBS) and incubated at 4 °C overnight. After washing with PBST, the plates were blocked with 200 μL PBST + 2% (w/v) BSA for 1 h at RT. The ELISA protocols described above were used for detection and quantification of PCKS9 or (oxidized) LDL-specific antibody titers. Plasma samples collected 8 weeks post immunization were analyzed. The plasma samples were evaluated per animal or pooled by group and tested in triplicate.
Human ldl
Manufactured by Thermo Fisher Scientific
The Human LDL is a laboratory equipment product designed for the analysis and study of low-density lipoprotein (LDL) in human samples. It provides the necessary tools and instruments to facilitate the measurement and investigation of LDL levels.
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Lab products found in correlation
2 protocols using human ldl
1
Evaluating Antibody Specificity Against Protein Targets
2
Foam Cell Formation Assay
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GGA was added to the medium at a final concentration of 10 μM one day after plating the peritoneal macrophages (105 cells/well) into 24-well plates. One day later, oxLDL, prepared via incubation of human LDL (Thermo Fisher Scientific) in PBS (containing 20 μM CuSO4) at 37 °C for 24 h, and added to macrophages at a final concentration of 100 μg/mL; cells were incubated for 24 h more. Thereafter, cells were washed three times with PBS, fixed with 4% paraformaldehyde for 10 min, stained with Oil Red O, and observed under an optical microscope (Olympus CK40).
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