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Human Apolipoprotein B/ApoB Quantikine ELISA Kit

Catalog #: DAPB00
8 Citations Datasheet / COA / SDS
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DAPB00
Human Apolipoprotein B/ApoB ELISA Standard Curve
2 Images
Product Details
Procedure
Citations (8)
FAQs
Supplemental Products
Reviews
Summary Precision Linearity Data Examples Product Datasheets Preparation and Storage Background Related Research Areas

Human Apolipoprotein B/ApoB Quantikine ELISA Kit Summary

Assay Type
Solid Phase Sandwich ELISA
Format
96-well strip plate
Assay Length
4.5 hours
Sample Type & Volume Required Per Well
Serum (10 uL), EDTA Plasma (10 uL), Heparin Plasma (10 uL)
Sensitivity
9.97 ng/mL
Assay Range
39.1 - 2,500 ng/mL (Serum, EDTA Plasma, Heparin Plasma)
Specificity
Natural human ApoB
Cross-reactivity
< 0.5% cross-reactivity observed with available related molecules.< 50% cross-species reactivity observed with species tested.
Interference
No significant interference observed with available related molecules.

Product Summary

The Quantikine Human Apolipoprotein B/ApoB Immunoassay is a 4.5 hour solid phase ELISA designed to measure ApoB levels in serum and plasma.

Precision

Intra-Assay Precision (Precision within an assay) Three samples of known concentration were tested on one plate to assess intra-assay precision
Inter-Assay Precision (Precision between assays) Three samples of known concentration were tested in separate assays to assess inter-assay precision

Serum, EDTA Plasma, Heparin Plasma

Intra-Assay Precision Inter-Assay Precision
Sample 1 2 3 1 2 3
n 20 20 20 20 20 20
Mean (pg/mL) 284 654 1228 261 717 1375
Standard Deviation 15 33.6 48.1 27.4 61.2 90.5
CV% 5.3 5.1 3.9 10.5 8.5 6.6

Linearity

To assess the linearity of the assay, samples containing high concentrations of ApoB were serially diluted with Calibrator Diluent to produce samples with values within the dynamic range of the assay.
Human Apolipoprotein B/ApoB ELISA Linearity

Scientific Data

N/A Human Apolipoprotein B/ApoB ELISA Standard Curve View Larger

Human Apolipoprotein B/ApoB ELISA Standard Curve

Product Datasheets

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Preparation and Storage

Shipping
The product is shipped with polar packs. Upon receipt, store it immediately at the temperature recommended below.
Storage
Store the unopened product at 2 - 8 °C. Do not use past expiration date.

Background: Apolipoprotein B/ApoB

The apolipoproteins are a structurally-unrelated group of proteins that have some association with the transport of lipids in blood. Apolipoproteins, plus phospholipids, cholesterol and triglycerides, form spherical particles with a lipid/hydrophobic center and a (apolipo)protein coat. The apolipoprotein coat promotes aqueous solubility and serves as a ligand for lipoprotein receptors. HDL may contain apolipoproteins A, C, D, E, J, L and M, while LDL contains apolipoproteins B and E.

ApoAI and ApoA2 are major protein components of serum high-density lipoprotein (HDL) and are produced by the liver and small intestine. They are involved in reverse cholesterol transport from tissues to the liver. Polymorphisms of ApoA2 are associated with disorders of cholesterol and fatty acid metabolism. Human ApoB (Apolipoprotein B-100) is a 550 kDa, secreted, palmitoylated glycoprotein that is part of LDL and VLDL particles. It is made by liver and is 4536 aa in length. It binds LDL to the ApoB/E receptor. ApoC activates lipoprotein lipase and may self-associate to form amyloid-type fibrils.

ApoE is a 34 kDa protein component of serum chylomicrons, VLDL, and HDL particles. It mediates the binding, uptake, and catabolism of these particles through interactions with the ApoE receptor and LDL receptors in the liver and brain. ApoE is important in fatty acid homeostasis and memory formation. Polymorphisms encode three variants (ApoE2, 3, 4) which are differentially related to the development of atherosclerosis and neurogenerative disorders, particularly Alzheimer's disease.

Serum amyloid A proteins (SAAs) are a family of homologous apolipoproteins of high density lipoprotein (HDL). They can be divided into two groups. The first group consists of the acute phase SAA1 and SAA2 that associate with HDL during inflammation and remodel the HDL particle by displacing apolipoprotein A1. The second group consists of constitutively expressed SAA4 and SAA5 that exist as minor apolipoproteins on HDL but make up more than 90% of the total SAA during homeostasis.

Entrez Gene IDs:
338 (Human); 238055 (Mouse); 54225 (Rat)
Alternate Names:
Apo B-100; APOB; ApoB-100; ApoB-48; apolipoprotein B (including Ag(x) antigen); Apolipoprotein B; apolipoprotein B-100; apolipoprotein B48; FLDB; LDLCQ4; mutant Apo B 100
⚠ WARNING: This product can expose you to chemicals including N,N-Dimethylforamide, which is known to the State of California to cause cancer. For more information, go to www.P65Warnings.ca.gov.

Assay Procedure

Refer to the product for complete assay procedure.
 
Bring all reagents and samples to room temperature before use. It is recommended that all samples, standards, and controls be assayed in duplicate.
  1. Prepare all reagents, standard dilutions, and samples as directed in the product insert.
  2. Remove excess microplate strips from the plate frame, return them to the foil pouch containing the desiccant pack, and reseal.

200 µL Assay Diluent

  1. Add 200 µL of Assay Diluent to each well.

50 µL Standard, Control, or Sample

  1. Add 50 µL of Standard, control, or sample to each well. Cover with a plate sealer, and incubate at room temperature for 2 hours on a horizontal orbital microplate shaker.
  2. Aspirate each well and wash, repeating the process 2 times for a total of 3 washes.

200 µL Conjugate

  1. Add 200 µL of Conjugate to each well. Cover with a new plate sealer, and incubate at room temperature for 2 hours on the shaker.
  2. Aspirate and wash 3 times.

200 µL Substrate Solution

  1. Add 200 µL Substrate Solution to each well. Incubate at room temperature for 30 minutes on the benchtop. PROTECT FROM LIGHT.

50 µL Stop Solution

  1. Add 50 µL of Stop Solution to each well. Read at 450 nm within 30 minutes. Set wavelength correction to 540 nm or 570 nm.

Citations for Human Apolipoprotein B/ApoB Quantikine ELISA Kit

R&D Systems personnel manually curate a database that contains references using R&D Systems products. The data collected includes not only links to publications in PubMed, but also provides information about sample types, species, and experimental conditions.

8 Citations: Showing 1 - 8
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  1. Caffeine blocks SREBP2-induced hepatic PCSK9 expression to enhance LDLR-mediated cholesterol clearance
    Authors: PF Lebeau, JH Byun, K Platko, P Saliba, M Sguazzin, ME MacDonald, G Paré, GR Steinberg, LJ Janssen, SA Igdoura, MA Tarnopolsk, SR Wayne Chen, NG Seidah, J Magolan, RC Austin
    Nature Communications, 2022;13(1):770.
    Species: Human
    Sample Types: Serum
  2. Preclinical development and phase 1 trial of a novel siRNA targeting lipoprotein(a)
    Authors: MJ Koren, PM Moriarty, SJ Baum, J Neutel, M Hernandez-, HS Weintraub, M Florio, H Kassahun, S Melquist, T Varrieur, SM Haldar, W Sohn, H Wang, M Elliott-Da, BM Rock, T Pei, O Homann, J Hellawell, GF Watts
    Nature Medicine, 2022;0(0):.
    Species: Cynomolgus Monkey
    Sample Types: Serum
  3. Effects of gastric bypass surgery on postprandial gut and systemic lipid handling
    Authors: P Jegatheesa, K Seyssel, N Stefanoni, V Rey, P Schneiter, V Giusti, V Lecoultre, L Tappy
    Clin Nutr ESPEN, 2020;35(0):95-102.
    Species: Human
    Sample Types: Plasma
  4. Glycan Node Analysis of Plasma-Derived Extracellular Vesicles
    Authors: SA Walker, JS Aguilar Dí, S Busatto, GA Wurtz, AC Zubair, CR Borges, J Wolfram
    Cells, 2020;9(9):.
    Species: Human
    Sample Types: Plasma
  5. Glycan Node Analysis of Plasma-Derived Extracellular Vesicles
    Authors: SA Walker, JS Aguilar Dí, S Busatto, GA Wurtz, AC Zubair, CR Borges, J Wolfram
    Cells, 2020;9(9):.
    Species: Human
    Sample Types: Plasma
  6. Differences in plasma fibrin clot composition in patients with thrombotic antiphospholipid syndrome compared with venous thromboembolism
    Authors: A Stachowicz, M Zabczyk, J Natorska, M Suski, R Olszanecki, R Korbut, JR Wi?niewski, A Undas
    Sci Rep, 2018;8(1):17301.
    Species: Human
    Sample Types: Plasma
  7. Effect of the catechol-O-methyltransferase inhibitors tolcapone and entacapone on fatty acid metabolism in HepaRG cells
    Authors: D Grünig, A Felser, U Duthaler, J Bouitbir, S Krähenbühl
    Toxicol. Sci., 2018;0(0):.
    Species: Human
    Sample Types: Cell Culture Supernates
  8. Lowering Low-Density Lipoprotein Particles in Plasma Using Dextran Sulphate Co-Precipitates Procoagulant Extracellular Vesicles
    Authors: JW Wang, YN Zhang, SK Sze, SM van de Weg, F Vernooij, AH Schoneveld, SH Tan, HH Versteeg, L Timmers, CSP Lam, DPV de Kleijn
    Int J Mol Sci, 2017;19(1):.
    Species: Human
    Sample Types: Plasma

FAQs

  1. What forms of ApoB are detected in Human Apolipoprotein B/ApoB Quantikine ELISA Kit, Catalog # DAPB00?

    • The immunogen for both the capture and the detection in DAPB00 is ApoB100 (aa 1206-1413). Based on amino acid sequences the kit is likely to see ApoB100, B48, B42, B37, and B34. The ApoB29 form is truncated at aa1305 and doesn't encompass the entire region of the immunogen so we are uncertain if this form is detected. A naturally-derived human ApoB, not a recombinant, is used as the standard in this ELISA kit, so it is a mixture of naturally occurring forms. We have not verified by testing each form independently for recognition in the assay.

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