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Human Angiopoietin-like 3 Quantikine ELISA Kit

R&D Systems | Catalog # DANL30

R&D Systems
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Key Product Details

Assay Length

3.5 hours

Sample Type & Volume Required Per Well

Cell Culture Supernates (50 µL), Serum (10 µL), EDTA Plasma (10 µL), Heparin Plasma (10 µL)

Sensitivity

0.035 ng/mL

Assay Range

0.156-10 ng/mL (Cell Culture Supernates, Serum, EDTA Plasma, Heparin Plasma)
View all Angiopoietin-like Protein 3/ANGPTL3 ELISA Kits »

Product Summary for Human Angiopoietin-like 3 Quantikine ELISA Kit

The Quantikine Human ANGPTL3 Immunoassay is a 3.5 hour solid-phase ELISA designed to measure human ANGPTL3 in cell culture supernates, serum, and plasma. It contains Sf 21-expressed recombinant human ANGPTL3 and has been shown to accurately quantitate the recombinant factor. Results obtained using natural human ANGPTL3 showed linear curves that were parallel to the standard curves obtained using the Quantikine kit standards. These results indicate that this kit can be used to determine relative mass values for naturally occurring human ANGPTL3.

Product Specifications

Assay Type

Solid Phase Sandwich ELISA

Format

96-well strip plate

Measurement

Quantitative ELISA

Detection Method

Colorimetric - 450nm (TMB)

Conjugate

HRP

Species

Human

Specificity

Natural and recombinant human ANGPTL3

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.

Sample Values

Serum/Plasma - Samples from apparently healthy volunteers were evaluated for the presence of human ANGPTL3 in this assay. No medical histories were available for the donors used in this study.

Sample TypeMean (ng/mL)Range (ng/mL)Standard Deviation (ng/mL)
Serum (n=36)94.846.1-15729.8
EDTA plasma (n=36)10855.8-17229.7
Heparin plasma (n=36)94.147.7-19428.7

Cell Culture Supernates - HepG2 human hepatocellular carcinoma cells were cultured in DMEM supplemented with 10% fetal bovine serum, 2 mM L-glutamine, 100 U/mL penicillin, and 100 μg/mL streptomycin sulfate until confluent. An aliquot of the cell culture supernate was removed, assayed for human ANGPTL3, and measured 2.62 ng/mL.

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.

Cell Culture Supernates, EDTA Plasma, Heparin Plasma, Serum

Intra-Assay Precision Inter-Assay Precision
Sample 1 2 3 1 2 3
n 20 20 20 20 20 20
Mean (ng/mL) 1.05 3.36 6.80 1.02 3.18 6.74
Standard Deviation 0.04 0.09 0.18 0.07 0.20 0.60
CV% 4.1 2.7 2.7 6.7 7.2 8.5

Recovery for Human Angiopoietin-like 3 Quantikine ELISA Kit

The recovery of ANGPTL3 spiked to levels throughout the range of the assay in various matrices was evaluated.

Sample Type Average % Recovery Range %
Cell Culture Media (n=4) 103 99-107
EDTA Plasma (n=4) 102 85-120
Heparin Plasma (n=4) 96 85-109
Serum (n=4) 96 81-115

Linearity

To assess the linearity of the assay, samples containing and/or spiked with high concentrations of ANGPTL3 were serially diluted with Calibrator Diluent to produce samples with values within the dynamic range of the assay.

Human Angiopoietin-like Protein 3/ANGPTL3 ELISA Linearity

Scientific Data Images for Human Angiopoietin-like 3 Quantikine ELISA Kit

Human Angiopoietin-like Protein 3/ANGPTL3 ELISA Standard Curve

Human Angiopoietin-like Protein 3/ANGPTL3 ELISA Standard Curve

Preparation and Storage

Shipping

The product is shipped at ambient temperature. Upon receipt, store it immediately at the temperature recommended below.

Stability & Storage

Store the unopened product at 2 - 8 °C. Do not use past expiration date.

Background: Angiopoietin-like Protein 3/ANGPTL3

Angiopoietin-like Protein 3 (ANGPTL3) is a secreted glycoprotein that is primarily expressed in the liver. Full length ANGPTL3 circulates in the plasma as do the proteolytically separated N- and C-terminal fragments containing the coiled coil domain and fibrinogen-like domains, respectively. ANGPTL3 directly inhibits lipoprotein lipase (LPL) and endothelial lipase (EL), enzymes responsible for hydrolyzing circulating triglycerides and HDL phospholipids. It promotes an increase in circulating triglyceride levels without altering VLDL or HDL secretion or uptake. ANGPTL3 is upregulated by LXR agonists and downregulated by Insulin, Leptin, and agonists of TR beta or PPAR beta. Its fibrinogen-like domain interacts with Integrin alphaV beta3 to induce endothelial cell adhesion, migration, and neovascularization. ANGPTL3 also promotes the expansion of hematopoietic stem cells.

Alternate Names

AGNPT5, Ang-5, Angiopoietin-5, ANGPTL3

Entrez Gene IDs

27329 (Human); 30924 (Mouse); 102136264 (Cynomolgus Monkey)

Gene Symbol

ANGPTL3

Additional Angiopoietin-like Protein 3/ANGPTL3 Products

Product Documents for Human Angiopoietin-like 3 Quantikine ELISA Kit

Certificate of Analysis

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Note: Certificate of Analysis not available for kit components.

Product Specific Notices for Human Angiopoietin-like 3 Quantikine ELISA Kit

For research use only

⚠ 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.

Citations for Human Angiopoietin-like 3 Quantikine ELISA Kit

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  • Human Angiopoietin-like 3 Quantikine ELISA Kit
    Name: Anonymous
    Sample Tested: Serum
    Verified Customer | Posted 08/09/2023
    Human Angiopoietin-like 3 Quantikine ELISA Kit DANL30
  • Human Angiopoietin-like 3 Quantikine ELISA Kit
    Name: Anonymous
    Sample Tested: Serum
    Verified Customer | Posted 05/26/2022
    Human Angiopoietin-like 3 Quantikine ELISA Kit DANL30

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Protocols

View specific protocols for Human Angiopoietin-like 3 Quantikine ELISA Kit (DANL30):

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.
    3. 100 µL Assay Diluent
  3.   Add 100 µL of Assay Diluent to each well.
    4. 50 µL Standard, Control, or Sample
  4.   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.
  5.   Aspirate each well and wash, repeating the process 3 times for a total of 4 washes.
    6. 200 µL Conjugate
  6.   Add 200 µL of Conjugate to each well. Cover with a new plate sealer, and incubate at room temperature for 1 hour on the shaker.
  7.   Aspirate and wash 4 times.
    8. 200 µL Substrate Solution
  8.   Add 200 µL Substrate Solution to each well. Incubate at room temperature for 30 minutes on the benchtop. PROTECT FROM LIGHT.
    9. 50 µL Stop Solution
  9.   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.

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FAQs

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