Use at an assay dependent concentration. Can be paired for Sandwich ELISA with Rabbit monoclonal [PSH24-26] to Human/Monkey Apolipoprotein CIII (Capture) (HA724433) and Recombinant Human Apolipoprotein CIII protein (HA211274) as the standard. The reference range value is 39.1-10,000 pg/mL.
ELISA
Use at an assay dependent concentration.
Target
Function
Apolipoprotein C-III also known as apo-CIII, and apolipoprotein C3, is a protein that in humans is encoded by the APOC3 gene. Apo-CIII is secreted by the liver as well as the small intestine, and is found on triglyceride-rich lipoproteins such as chylomicrons, very low density lipoprotein (VLDL), and remnant cholesterol. APOC3 inhibits lipoprotein lipase and hepatic lipase; it is thought to inhibit hepatic uptake of triglyceride-rich particles. The APOA1, APOC3 and APOA4 genes are closely linked in both rat and human genomes. The A-I and A-IV genes are transcribed from the same strand, while the A-1 and C-III genes are convergently transcribed. An increase in apoC-III levels induces the development of hypertriglyceridemia. Recent evidence suggests an intracellular role for Apo-CIII in promoting the assembly and secretion of triglyceride-rich VLDL particles from hepatic cells under lipid-rich conditions. However, two naturally occurring point mutations in human apoC3 coding sequence, namely Ala23Thr and Lys58Glu have been shown to abolish the intracellular assembly and secretion of triglyceride-rich VLDL particles from hepatic cells.
Background References
1. Naber A et al. Apolipoprotein-CIII O-Glycosylation, a Link between GALNT2 and Plasma Lipids. Int J Mol Sci. 2023 Oct
2. Naber A et al. Apolipoprotein-CIII O-Glycosylation Is Associated with Micro- and Macrovascular Complications of Type 2 Diabetes. Int J Mol Sci. 2024 May
Sandwich ELISA analysis of Human Apolipoprotein CIII matched pair antibodies
Capture: HA724433, Human/Monkey Apolipoprotein CIII Rabbit mAb [PSH24-26] Detector: HA724434, Human/Monkey Apolipoprotein CIII Rabbit mAb [PSH24-27]
Elisa assay was performed by coating wells of a 96-well plate with 100 µl per well of capture antibody (HA724433) diluted in carbonate/bicarbonate buffer, at a concentration of 2 μg/ml overnight at 4℃. Wells of the plate were washed, blocked with 150 µl 0.05% tween-20 1% BSA blocking buffer, and incubated with serial diluted Recombinant Human Apolipoprotein CIII protein (HA211274) starting from 10,000 pg/ml to 0 pg/ml and detect antibody (HA724434, Biotin, 0.2 µg/ml) for 1 hour at 30℃ with shaking. Then the plate was washed and incubated with 100 µl per well of SA-HRP for 0.5 hour at 30℃ with shaking. Detection was performed using an Ultra TMB Substrate for 10 minutes at room temperature in the dark. The reaction was stopped with sulfuric acid and absorbances were read on a spectrophotometer at 450 nm.
Interpolated concentrations of native Apolipoprotein CIII in HepG2 cell culture supernatant.
Capture: HA724433, Human/Monkey Apolipoprotein CIII Rabbit mAb [PSH24-26] Detector: HA724434, Human/Monkey Apolipoprotein CIII Rabbit mAb [PSH24-27]
The concentrations of Apolipoprotein CIII were measured in duplicates, interpolated from the Apolipoprotein CIII standard curve and corrected for sample dilution. Undiluted samples are HepG2 cell culture supernatant 50%. The interpolated dilution factor corrected values are plotted (mean +/- SD, n=2). The mean Apolipoprotein CIII concentration was determined to be 2,809 pg/ml in HepG2 cell culture supernatant.
Interpolated concentrations of native Apolipoprotein CIII in human serum samples.
Capture: HA724433, Human/Monkey Apolipoprotein CIII Rabbit mAb [PSH24-26] Detector: HA724434, Human/Monkey Apolipoprotein CIII Rabbit mAb [PSH24-27]
The concentrations of Apolipoprotein CIII were measured in duplicates, interpolated from the Apolipoprotein CIII standard curve and corrected for sample dilution. Undiluted samples are human serum 0.005%. The interpolated dilution factor corrected values are plotted (mean +/- SD, n=2). The mean Apolipoprotein CIII concentration was determined to be 71.3 μg/ml in human serum.
Interpolated concentrations of native Apolipoprotein CIII in Rhesus monkey serum samples.
Capture: HA724433, Human/Monkey Apolipoprotein CIII Rabbit mAb [PSH24-26] Detector: HA724434, Human/Monkey Apolipoprotein CIII Rabbit mAb [PSH24-27]
The concentrations of Apolipoprotein CIII were measured in duplicates, interpolated from the Apolipoprotein CIII standard curve and corrected for sample dilution. Undiluted samples are Rhesus monkey serum 2.5%. The interpolated dilution factor corrected values are plotted (mean +/- SD, n=2). The mean Apolipoprotein CIII concentration was determined to be 191.5 ng/ml in Rhesus monkey serum.
Interpolated concentrations of spiked Apolipoprotein CIII in human cell culture media samples.
Capture: HA724433, Human/Monkey Apolipoprotein CIII Rabbit mAb [PSH24-26] Detector: HA724434, Human/Monkey Apolipoprotein CIII Rabbit mAb [PSH24-27]
The concentrations of Apolipoprotein CIII were measured in duplicates, interpolated from the Apolipoprotein CIII standard curves and corrected for sample dilution. Diluted samples are as follows: 50% cell culture media with FBS. The interpolated dilution factor corrected values are plotted (mean +/- SD, n=2).
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