Human ApoCIII ELISA Kit, Trial Size
Contents
1. Product Manual Human ApoClll ELISA Kit Trial Size Catalog Number STA 366 T 32 assays FOR RESEARCH USE ONLY Not for use in diagnostic procedures CELL BIOLABS INC Creating Solutions for Life Science Research Introduction Lipoproteins are submicroscopic particles composed of lipid and protein held together by noncovalent forces Their general structure is that of a putative spheroidal microemulsion formed from an outer layer of phospholipids unesterified cholesterol and proteins with a core of neutral lipids predominately cholesteryl esters and triacylglycerols TAG Plasma apolipoproteins can be grouped into two classes the nonexchangeable apolipoproteins ApoB 100 and ApoB 48 and the exchangeable apolipoproteins ApoAI ApoAII ApoAIV ApoCI ApoCII ApoCHI and ApoE ApoCIlll is a 79 amino acid protein synthesized by liver and intestine which is an essential constituent of circulating particles rich in triacylglycerol i e chylomicrons and VLDLs ApoCHI inhibits the hydrolysis of TG rich particles by the lipoprotein lipase and their hepatic uptake mediated by ApoE Cholesterol Triacyl Phospholipid glycerol Figure 1 Structure of VLDL Cell Biolabs Human ApoCIII ELISA Kit is an enzyme immunoassay developed for the detection and quantitation of human ApoCIII in plasma serum or other biological fluid samples The kit has detection sensitivity limit of 50 pg mL human ApoCIII This Trial Size kit provides suf2. 1500 2000 2500 Human ApoClll pg mL Figure 2 Human ApoCIII ELISA Standard Curve References 1 Segrest J P Garber D W Brouillette C G Harvey S C and Anantharamaiah G M Adv Protein Chem 45 303 369 1994 2 Segrest J P Jones M K De Loof H Brouillette C G Venkatachalapathi Y V and Anantharamaiah G M J Lipid Res 33 141 166 1992 3 Mendivil C O Zheng C Furtado J Lel J and Sacks F M Arteriosclerosis Thrombosis and Vascular Biology 30 2 239 45 2009 Warranty These products are warranted to perform as described in their labeling and in Cell Biolabs literature when used in accordance with their instructions THERE ARE NO WARRANTIES THAT EXTEND BEYOND THIS EXPRESSED WARRANTY AND CELL BIOLABS DISCLAIMS ANY IMPLIED WARRANTY OF MERCHANTABILITY OR WARRANTY OF FITNESS FOR PARTICULAR PURPOSE CELL BIOLABS sole obligation and purchaser s 6 j J _ CELL BIOLABS INC e exclusive remedy for breach of this warranty shall be at the option of CELL BIOLABS to repair or replace the products In no event shall CELL BIOLABS be liable for any proximate incidental or consequential damages in connection with the products Contact Information Cell Biolabs Inc 7758 Arjons Drive San Diego CA 92126 Worldwide 1 858 271 6500 USA Toll Free 1 888 CBL 0505 E mail tech cellbiolabs com www cellbiolabs com 2013 2015 Cell Biolabs Inc All rights reserved No part of these works may b
3. a Collect blood with heparin or EDTA and centrifuge for 10 minutes at 1000 g at 4 C Remove the plasma and assay immediately or store samples at 80 C up to three months Normal plasma samples require at least 20 000 fold dilution or more with PBS containing 0 1 BSA immediately before running the ELISA A sample titration may be performed to ensure sample OD values fall within the middle range of the standard curve e Serum Harvest serum and centrifuge for 10 minutes at 1000 g at 4 C Assay immediately or store samples at 80 C up to three months Normal serum samples require at least 20 000 fold dilution or more with PBS containing 0 1 BSA immediately before running the ELISA A sample titration may be performed to ensure sample OD values fall within the middle range of the standard curve o gt CELL BIOLABS INC a Other Biological Fluids Centrifuge samples for 10 minutes at 1000 g at 4 C Assay immediately or store samples at 80 C up to three months Assay Protocol l 2a 10 11 Prepare dilutions of plasma serum or other biological fluid samples in PBS containing 0 1 BSA Add 100 uL of ApoCI unknown sample or standard to the Anti ApoCII Antibody Coated Plate Each ApoCHI unknown sample standard and blank should be assayed in duplicate Incubate at 37 C for at least 2 hours or 4 C overnight Wash microwell strips 3 times with 250 uL 1X Wash Buffer per well with thorough aspiration between each wash Afte
4. e reproduced in any form without permissions in writing eo CELL BIOLABS INC AN
5. ficient reagents to perform up to 32 assays including standard curve and unknown samples N CELL BIOLABS INC Related Products 1 CoN DAMA AWN STA 137 Rabbit Anti Human Apolipoprotein CIII Polyclonal Antibody STA 214 Copper Cu Oxidized Human Low Density Lipoprotein LDL STA 237 Human Apolipoprotein CHI STA 362 Human ApoAI ELISA Kit STA 363 Human ApoAII ELISA Kit STA 364 Human ApoCI ELISA Kit STA 365 Human ApoCII ELISA Kit STA 367 Human ApoE ELISA Kit STA 368 Human ApoB 100 ELISA Kit 10 STA 369 Human Oxidized LDL ELISA Kit Kit Components Box 1 shipped at room temperature 1 SD Pm eM Anti ApoCIII Antibody Coated Plate Part No 236601 T One strip well plate containing 32 wells 8 x 4 Biotinylated Anti ApoCHI Antibody 1000X Part No 236602 T One 10 uL vial Streptavidin Enzyme Conjugate Part No 310803 T One 10 uL vial Assay Diluent Part No 310804 T One 20 mL bottle 10X Wash Buffer Part No 310806 T One 30 mL bottle Substrate Solution Part No 310807 T One 4 mL amber bottle Stop Solution Part No 310808 T One 4 mL bottle Box 2 shipped on blue ice packs 1 Human ApoCH Standard Part No 236603 T One 20 uL vial of 1 ug mL Human ApoCIII in PBS plus BSA Materials Not Supplied SY Oe eS oe Plasma Serum or Other Biological Fluids PBS containing 0 1 BSA 10 uL to 1000 uL adjustable single channel micropipettes with disposable tips 50 uL
6. r the last wash empty wells and tap microwell strips on absorbent pad or paper towel to remove excess 1X Wash Buffer Add 100 uL of the diluted Biotinylated Anti ApoCIII antibody to each well Incubate at room temperature for 1 hour on an orbital shaker Wash the strip wells 3 times according to step 4 above Add 100 uL of the diluted Streptavidin Enzyme Conjugate to each well Incubate at room temperature for 1 hour on an orbital shaker Wash the strip wells 3 times according to step 4 above Proceed immediately to the next step Warm Substrate Solution to room temperature Add 100 uL of Substrate Solution to each well including the blank wells Incubate at room temperature on an orbital shaker Actual incubation time may vary from 2 30 minutes Note Watch plate carefully if color changes rapidly the reaction may need to be stopped sooner to prevent saturation Stop the enzyme reaction by adding 100 uL of Stop Solution into each well including the blank wells Results should be read immediately color will fade over time Read absorbance of each microwell on a spectrophotometer using 450 nm as the primary wave length eo CELL BIOLABS INC a Example of Results The following figures demonstrate typical results with the Human ApoCIH ELISA Kit One should use the data below for reference only This data should not be used to interpret actual results m co E c S iN t a O 1000
7. to 300 uL adjustable multichannel micropipette with disposable tips Multichannel micropipette reservoir Microplate reader capable of reading at 450 nm 620 nm as optional reference wave length CELL BIOLABS INC ae Storage Upon receipt aliquot and store the Human ApoCIII Standard at 20 C to avoid multiple freeze thaw cycles Store all other components at 4 C Preparation of Reagents e 1X Wash Buffer homogeneity e Biotinylated Anti ApoCII Antibody and Streptavidin Enzyme Conjugate Immediately before use dilute the Biotinylated Anti ApoCIII antibody 1 1000 and the Streptavidin Enzyme Conjugate Dilute the 10X Wash Buffer Concentrate to 1X with deionized water 1 1000 with Assay Diluent Do not store diluted solutions Preparation of Human ApoCIII Standard Prepare a dilution series of human ApoCIII standards in the concentration range of 0 to 2 000 pg mL in Assay Diluent Table 1 Human Standard 1 pg mL Human ApoCIII Assay Diluent ApoClilI Tubes Standard uL uL pg mL 1 4 1998 2000 2 500 of Tube 1 500 1000 3 500 of Tube 2 500 500 4 500 of Tube 3 500 250 5 500 of Tube 4 500 125 6 500 of Tube 5 500 62 5 7 500 of Tube 6 500 31 3 8 0 500 0 Table 1 Preparation of Human ApoCIII Standards Preparation of Samples The following recommendations are only guidelines and may be altered to optimize or complement the user s experimental design e Plasm
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