Human APOA1 ELISA Kit EZ-SetTM (DIY Antibody Pairs)

Apolipoprotein A1 is a protein that in humans is encoded by the APOA1 gene. This gene encodes apolipoprotein A-I, which is the major protein component of high density lipoprotein (HDL) in plasma. The encoded preproprotein is proteolytically processed to generate the mature protein, which promotes cholesterol efflux from tissues to the liver for excretion, and is a cofactor for lecithin cholesterolacyltransferase (LCAT), an enzyme responsible for the formation of most plasma cholesteryl esters. This gene is closely linked with two other apolipoprotein genes on chromosome 11. Defects in this gene are associated with HDL deficiencies, including Tangier disease, and with systemic non-neuropathic amyloidosis. Alternative splicing results in multiple transcript variants, at least one of which encodes a preproprotein.

Lots Lot 1 (pg/ml) Lot 2 (pg/ml) Lot 3 (pg/ml) Lot 4 (pg/ml) Mean (pg/ml) Standard Deviation CV (%) Sample 1 4918 4239 5307 3555 4505 336.96 7.5% Sample 2 18666 19845 16500 24262 19818 400.33 2.0% Sample 3 98104 76874 81171 83329 84870 5236.45 6.2% Lots Lot 1 (pg/ml) Lot 2 (pg/ml) Lot 3 (pg/ml) Lot 4 (pg/ml) Mean (pg/ml) Standard Deviation CV (%) Lots Lot 1 (pg/ml) Lot 2 (pg/ml) Lot 3 (pg/ml) Lot 4 (pg/ml) Mean (pg/ml) Standard Deviation CV (%) Sample 1 4918 4239 5307 3555 4505 336.96 7.5% Sample 1 4918 4239 5307 3555 4505 336.96 7.5% Sample 2 18666 19845 16500 24262 19818 400.33 2.0% Sample 2 18666 19845 16500 24262 19818 400.33 2.0% Sample 3 98104 76874 81171 83329 84870 5236.45 6.2% Sample 3 98104 76874 81171 83329 84870 5236.45 6.2%

Apolipoprotein A-I; Apo-AI; ApoA-I; Apolipoprotein A1; Proapolipoprotein A-I; ProapoA-I; Truncated apolipoprotein A-I; Apolipoprotein A-I (1-242) ; APOA1

Major protein of plasma HDL, also found in chylomicrons. Synthesized in the liver and small intestine. The oxidized form at Met-110 and Met-136 is increased in individuals with increased risk for coronary artery disease, such as in carrier of the eNOSa/b genotype and exposure to cigarette smoking. It is also present in increased levels in aortic lesions relative to native ApoA-I and increased levels are seen with increasing severity of disease.

Cardiovascular, Cholesterol Metabolism, Lipid and Lipoprotein Metabolism, Lipid Metabolism, Lipids/Lipoproteins, Metabolic Signaling Pathways, Metabolism, Pathways and Processes, Signal Transduction

Intra-Assay Precision Inter-Assay Precision Sample 1 2 3 1 2 3 n 16 16 16 24 24 24 Mean (pg/ml) 4918 18666 98104 5841 22667 77833 Standard deviation 155.90 1472.75 5660.60 238.90 988.28 5168.11 CV (%) 3.2% 7.9% 5.8% 4.1% 4.4% 6.6% Intra-Assay Precision Inter-Assay Precision Sample 1 2 3 1 2 3 n 16 16 16 24 24 24 Mean (pg/ml) 4918 18666 98104 5841 22667 77833 Standard deviation 155.90 1472.75 5660.60 238.90 988.28 5168.11 CV (%) 3.2% 7.9% 5.8% 4.1% 4.4% 6.6% Intra-Assay Precision Inter-Assay Precision Intra-Assay Precision Inter-Assay Precision Sample 1 2 3 1 2 3 Sample 1 2 3 1 2 3 n 16 16 16 24 24 24 n 16 16 16 24 24 24 Mean (pg/ml) 4918 18666 98104 5841 22667 77833 Mean (pg/ml) 4918 18666 98104 5841 22667 77833 Standard deviation 155.90 1472.75 5660.60 238.90 988.28 5168.11 Standard deviation 155.90 1472.75 5660.60 238.90 988.28 5168.11 CV (%) 3.2% 7.9% 5.8% 4.1% 4.4% 6.6% CV (%) 3.2% 7.9% 5.8% 4.1% 4.4% 6.6%

Participates in the reverse transport of cholesterol from tissues to the liver for excretion by promoting cholesterol efflux from tissues and by acting as a cofactor for the lecithin cholesterol acyltransferase (LCAT) . As part of the SPAP complex, activates spermatozoa motility.

Catalog number Description Quantity Storage of opened/reconstituted material EZ1456-CA Rabbit anti- human APOA1 polyclonal antibody (Capture Antibody) 500 μl, 0.4 mg/mL (recommended dilution 1:100) Store undiluted at 4°C for 1 month or at -20°C for 3 months provided this is within the expiration date of the kit. EZ1456-DA Biotinylated goat anti- human APOA1 polyclonal antibody (Detection Antibody) 500 μl, 33 μg/mL (recommended dilution 1:100) AR1103 Avidin-Biotin-Peroxidase Complex (ABC) 500 μl (recommended dilution 1:100) EK1456-ST Lyophilized recombinant human APOA1 standard 200 ng/tubex3 Discard the standard stock solution after 12 hours at 4°C. May be stored at -20°C for 48 hours provided this is within the expiration date of the kit. Catalog number Description Quantity Storage of opened/reconstituted material EZ1456-CA Rabbit anti- human APOA1 polyclonal antibody (Capture Antibody) 500 μl, 0.4 mg/mL (recommended dilution 1:100) Store undiluted at 4°C for 1 month or at -20°C for 3 months provided this is within the expiration date of the kit. EZ1456-DA Biotinylated goat anti- human APOA1 polyclonal antibody (Detection Antibody) 500 μl, 33 μg/mL (recommended dilution 1:100) AR1103 Avidin-Biotin-Peroxidase Complex (ABC) 500 μl (recommended dilution 1:100) EK1456-ST Lyophilized recombinant human APOA1 standard 200 ng/tubex3 Discard the standard stock solution after 12 hours at 4°C. May be stored at -20°C for 48 hours provided this is within the expiration date of the kit. Catalog number Description Quantity Storage of opened/reconstituted material Catalog number Description Quantity Storage of opened/reconstituted material EZ1456-CA Rabbit anti- human APOA1 polyclonal antibody (Capture Antibody) 500 μl, 0.4 mg/mL (recommended dilution 1:100) Store undiluted at 4°C for 1 month or at -20°C for 3 months provided this is within the expiration date of the kit. EZ1456-CA Rabbit anti- human APOA1 polyclonal antibody (Capture Antibody) 500 μl, 0.4 mg/mL (recommended dilution 1:100) Store undiluted at 4°C for 1 month or at -20°C for 3 months provided this is within the expiration date of the kit. EZ1456-DA Biotinylated goat anti- human APOA1 polyclonal antibody (Detection Antibody) 500 μl, 33 μg/mL (recommended dilution 1:100) EZ1456-DA Biotinylated goat anti- human APOA1 polyclonal antibody (Detection Antibody) 500 μl, 33 μg/mL (recommended dilution 1:100) AR1103 Avidin-Biotin-Peroxidase Complex (ABC) 500 μl (recommended dilution 1:100) AR1103 Avidin-Biotin-Peroxidase Complex (ABC) 500 μl (recommended dilution 1:100) EK1456-ST Lyophilized recombinant human APOA1 standard 200 ng/tubex3 Discard the standard stock solution after 12 hours at 4°C. May be stored at -20°C for 48 hours provided this is within the expiration date of the kit. EK1456-ST Lyophilized recombinant human APOA1 standard 200 ng/tubex3 Discard the standard stock solution after 12 hours at 4°C. May be stored at -20°C for 48 hours provided this is within the expiration date of the kit.

Store at 4°C for 6 months, at -20°C for 12 months. Avoid multiple freeze-thaw cycles (Ships with gel ice, can store for up to 3 days in room temperature. Freeze upon receiving.)

Concentration (ng/ml) 0 3.12 6.25 12.5 25 50 100 200 O.D. 0.009 0.13 0.228 0.425 0.819 1.261 1.894 2.41 Concentration (ng/ml) 0 3.12 6.25 12.5 25 50 100 200 O.D. 0.009 0.13 0.228 0.425 0.819 1.261 1.894 2.41 Concentration (ng/ml) 0 3.12 6.25 12.5 25 50 100 200 Concentration (ng/ml) 0 3.12 6.25 12.5 25 50 100 200 O.D. 0.009 0.13 0.228 0.425 0.819 1.261 1.894 2.41 O.D. 0.009 0.13 0.228 0.425 0.819 1.261 1.894 2.41

1. Microplate reader in standard size. 2. Automated plate washer. 3. Incubator. 4. Adjustable pipettes and pipette tips. Multichannel pipettes are recommended in the condition of large amount of samples in the detection. 5. Clean tubes and Eppendorf tubes. 6. 96 well microplate (Cat# AR1100) 7. Plate Sealers. 8. Capture Antibody Diluent: PBS. 9. Reagent Diluent: 1% BSA in PBS, pH 7.2-7.4, 0.2 μm filtered. 10. Substrate Solution: Tetramethylbenzidine (Cat# AR1104) 11. Stop Solution: 2 N H2SO4 (Cat# AR1105) 12. Wash Buffer (PBS and PBS-T) . Recipes: PBS: 8g NaCl, 0.2g KCl, 1.15g Na2HPO4, 0.2g KH2PO4, adjust the total volume to 1 L with distilled water, pH 7.2-7.4, 0.2 μm filtered. PBS-T: 0.1% Tween® 20 in PBS, pH 7.2-7.4. *Item 6 – 12 are included in the EZ Set Accessory Kit (EZA001)