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Apolipoprotein A (APOA) ELISA Kit ELISA Kit

APOA Reactivity: Human Colorimetric Sandwich ELISA Serum
Pubmed (1)
Catalog No. ABIN2345022
$649.00
Plus shipping costs $45.00
96 tests
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Delivery in 2 to 3 Business Days
  • Target
    Reactivity
    Human
    Detection Method
    Colorimetric
    Method Type
    Sandwich ELISA
    Application
    ELISA
    Sample Type
    Serum, Plasma
    Analytical Method
    Quantitative
    Sensitivity
    1 ng/mL
    Characteristics
    Human Apo(a) ELISA Kit is an enzyme immunoassay developed for the detection and quantitation of human Apo(a) in plasma, serum or other biological fluid samples. The kit has detection sensitivity limit of 1 ng/mL human Apo(a). Each kit provides sufficient reagents to perform up to 96 assays including standard curve and unknown samples. 2
    Components
    1. Anti-Apo(a) Antibody Coated Plate : One 96-well strip plate (8 x 12).
    2. Biotinylated Anti-Apo(a) Antibody (1000X) : One 20 μL vial.
    3. Streptavidin-Enzyme Conjugate : One 20 μL vial.
    4. Assay Diluent : One 50 mL bottle.
    5. 10X Wash Buffer : One 100 mL bottle.
    6. Substrate Solution : One 12 mL amber bottle.
    7. Stop Solution (Part. No. 310808): One 12 mL bottle.

    Box 2 (shipped on blue ice packs)

    Material not included
    1. Plasma, Serum or Other Biological Fluids
    2. PBS containing 0.1 % BSA
    3. 10 μL to 1000 μL adjustable single channel micropipettes with disposable tips
    4. 50 μL to 300 μL adjustable multichannel micropipette with disposable tips
    5. Multichannel micropipette reservoir
    6. Microplate reader capable of reading at 450 nm (620 nm as optional reference wave length) 3
  • Target
    Alternative Name
    Apo(a) (APOA ELISA Kit Abstract)
    Background
    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, ApoCIII, and ApoE). Lipoprotein(a) (Lp(a)) is a low density lipoprotein (LDL)-like particle synthesized by the liver that consists of ApoB-100 Molecule covalently linked to a very large glycoprotein known as apolipoprotein(a) (Apo(a)) (see Figure 1). Many epidemiological studies have reported positive associations of baseline Lp(a) concentration with coronary heart disease (CHD) risk. The Apo(a) chain contains five cysteine rich domains known as "kringles". The fourth kringle is homologous with the fibrin-binding domain of plasminogen, a plasma protein that dissolves blood clots when activated. Because of this structural similarity to plasminogen, Lp(a) interferes with fibrinolysis by competing with plasminogen binding to molecules and cells. This impairs plasminogen activation, plasmin generation, and fibrinolysis. Lp(a) also binds to macrophages via a high-affinity receptor that promotes foam cell formation and the deposition of cholesterol in atherosclerotic plaques. Figure 1: Schematic diagram of lipoprotein(a): Lipoprotein(a) consists of an LDL particle and a glycoprotein molecule, Apo(a), attached to the ApoB-100 moiety of the LDL particle through a disulfide bond.
  • Application Notes
    Optimal working dilution should be determined by the investigator.
    Comment

    • Quantify Apo(a) in plasma, serum or other biological fluid samples
    • Detection sensitivity limit of 1 ng/mL human Apo(a)
    • Human Apo(a) standard included

    Plate
    Pre-coated
    Reagent Preparation

    1X Wash Buffer: Dilute the 10X Wash Buffer Concentrate to 1X with deionized water. Stir to homogeneity. Biotinylated Anti-Apo(a) Antibody and Streptavidin-Enzyme Conjugate: Immediately before use dilute the Biotinylated Anti-Apo(a) antibody 1:1000 and the Streptavidin-Enzyme Conjugate 1:10,000 with Assay Diluent. Do not store diluted solutions.

    Sample Preparation

    The following recommendations are only guidelines and may be altered to optimize or complement the user's experimental design. Plasma: 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 about 1,000 to 10,000 fold dilution with PBS containing 0.1 % BSA immediately before running the ELISA. 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 about 1000 to 10,000 fold dilution with PBS containing 0.1 % BSA immediately before running the ELISA. 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. 4

    Assay Procedure
    1. Prepare dilutions of plasma, serum or other biological fluid samples in PBS containing 0.1 % BSA.
    2. Add 100 μL of Apo(a) unknown sample or standard to the Anti-Apo(a) Antibody Coated Plate. Each Apo(a) unknown sample, standard and blank should be assayed in duplicate.
    3. Incubate at 37 °C for at least 2 hours or 4 °C overnight.
    4. Wash microwell strips 3 times with 250 μL 1X Wash Buffer per well with thorough aspiration between each wash. After the last wash, empty wells and tap microwell strips on absorbent pad or paper towel to remove excess 1X Wash Buffer.
    5. Add 100 μL of the diluted Biotinylated Anti-Apo(a) antibody to each well. Incubate at room temperature for 1 hour on an orbital shaker.
    6. Wash the strip wells 3 times according to step 4 above.
    7. Add 100 μL of the diluted Streptavidin-Enzyme Conjugate to each well. Incubate at room temperature for 1 hour on an orbital shaker.
    8. Wash the strip wells 3 times according to step 4 above. Proceed immediately to the next step.
    9. Warm Substrate Solution to room temperature. Add 100 L 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.
    10. Stop the enzyme reaction by adding 100 μL of Stop Solution into each well, including the blank wells. Results should be read immediately (color will fade over time).
    11. Read absorbance of each microwell on a spectrophotometer using 450 nm as the primary wave length. 5
    Restrictions
    For Research Use only
  • Handling Advice
    Avoid multiple freeze/thaw cycles.
    Storage
    4 °C/-20 °C
    Storage Comment
    Upon receipt, aliquot and store the Human Apo(a) Standard at -20°C to avoid multiple freeze/thaw cycles. Store all other components at 4°C.
  • Bissig-Choisat, Wang, Legras, Saha, Chen, Bell, Pankowicz, Hill, Barzi, Leyton, Leung, Kruse, Himes, Goss, Wilson, Chan, Lagor, Bissig: "Development and rescue of human familial hypercholesterolaemia in a xenograft mouse model." in: Nature communications, Vol. 6, pp. 7339, 2015 (PubMed).

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