Human Angiopoietin-like 4 DuoSet ELISA

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Ancillary Products Available
Human Angiopoietin-like Protein 4 / ANGPTL4 ELISA Standard Curve
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Product Details
Citations (22)
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Human Angiopoietin-like 4 DuoSet ELISA Summary

Assay Type
Solid Phase Sandwich ELISA
96-well strip plate
Sample Volume Required
100 µL
Sufficient Materials
For fifteen 96-well plates*
Please see the product datasheet

* Provided that the recommended microplates, buffers, diluents, substrates and solutions are used, and the assay is run as summarized in the Assay Procedure provided.

This DuoSet ELISA Development kit contains the basic components required for the development of sandwich ELISAs to measure natural and recombinant human Angiopoietin-like4. The suggested diluent is suitable for the analysis of most cell culture supernate samples. Diluents for complex matrices, such as serum and plasma, should be evaluated prior to use in this DuoSet.

Product Features

  • Optimized capture and detection antibody pairings with recommended concentrations save lengthy development time
  • Development protocols are provided to guide further assay optimization
  • Assay can be customized to your specific needs
  • Economical alternative to complete kits

Kit Content

  • Capture Antibody
  • Detection Antibody
  • Recombinant Standard
  • Streptavidin conjugated to horseradish-peroxidase (Streptavidin-HRP)

Other Reagents Required

DuoSet Ancillary Reagent Kit 2 (5 plates): (Catalog # DY008) containing 96 well microplates, plate sealers, substrate solution, stop solution, plate coating buffer (PBS), wash buffer, and Reagent Diluent Concentrate 2.

The components listed above may be purchased separately:

PBS: (Catalog # DY006), or 137 mM NaCl, 2.7 mM KCl, 8.1 mM Na2HPO4, 1.5 mM KH2PO4, pH 7.2 - 7.4, 0.2 µm filtered

Wash Buffer: (Catalog # WA126), or 0.05% Tween® 20 in PBS, pH 7.2-7.4

Reagent Diluent: (Catalog # DY995), or 1% BSA in PBS, pH 7.2-7.4, 0.2 µm filtered

Substrate Solution: 1:1 mixture of Color Reagent A (H2O2) and Color Reagent B (Tetramethylbenzidine) (Catalog # DY999)

Stop Solution: 2 N H2SO4 (Catalog # DY994)

Microplates: R&D Systems (Catalog # DY990)

Plate Sealers: ELISA Plate Sealers (Catalog # DY992)

Data Example

Human Angiopoietin-like Protein 4 / ANGPTL4 ELISA Standard Curve

Product Datasheets

Preparation and Storage

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 4/ANGPTL4

Angiopoietin-like, also known as angiopoietin-related proteins, are structurally related to the angiopoietins. They are secreted proteins with N-terminal coiled-coil domains and C-terminal fibrinogen-like domains. Unlike the angiopoietins, ANGPTLs do not bind Tie-2. Seven ANGPTL proteins have been identified (ANGPTL1 through 7).

Entrez Gene IDs:
51129 (Human); 57875 (Mouse); 362850 (Rat)
Alternate Names:
Angiopoietin like Protein 4; angiopoietin-like 4; Angiopoietin-like Protein 4; ANGPTL4; ARP4fasting-induced adipose factor; FIAF; FIAFhepatic angiopoietin-related protein; Hepatic fibrinogen/angiopoietin-related protein; HFARP; HFARPANGPTL2; NL2; peroxisome proliferator-activated receptor (PPAR) gamma inducedangiopoietin-related protein; PGAR; PGARangiopoietin-related protein 4; pp1158; PPARG angiopoietin related protein

Assay Procedure


Plate Preparation

  1. Dilute the Capture Antibody to the working concentration in PBS without carrier protein. Immediately coat a 96-well microplate with 100 μL per well of the diluted Capture Antibody. Seal the plate and incubate overnight at room temperature.
  2. Aspirate each well and wash with Wash Buffer, repeating the process two times for a total of three washes. Wash by filling each well with Wash Buffer (400 μL) using a squirt bottle, manifold dispenser, or autowasher. Complete removal of liquid at each step is essential for good performance. After the last wash, remove any remaining Wash Buffer by aspirating or by inverting the plate and blotting it against clean paper towels.
  3. Block plates by adding 300 μL Reagent Diluent to each well. Incubate at room temperature for a minimum of 1 hour.
  4. Repeat the aspiration/wash as in step 2. The plates are now ready for sample addition.

Assay Procedure

  1. Add 100 μL of sample or standards in Reagent Diluent, or an appropriate diluent, per well. Cover with an adhesive strip and incubate 2 hours at room temperature.
  2. Repeat the aspiration/wash as in step 2 of Plate Preparation.
  3. Add 100 μL of the Detection Antibody, diluted in Reagent Diluent, to each well. Cover with a new adhesive strip and incubate 2 hours at room temperature.
  4. Repeat the aspiration/wash as in step 2 of Plate Preparation.
  5. Add 100 μL of the working dilution of Streptavidin-HRP to each well. Cover the plate and incubate for 20 minutes at room temperature. Avoid placing the plate in direct light.
  6. Repeat the aspiration/wash as in step 2.
  7. Add 100 μL of Substrate Solution to each well. Incubate for 20 minutes at room temperature. Avoid placing the plate in direct light.
  8. Add 50 μL of Stop Solution to each well. Gently tap the plate to ensure thorough mixing.
  9. Determine the optical density of each well immediately, using a microplate reader set to 450 nm. If wavelength correction is available, set to 540 nm or 570 nm. If wavelength correction is not available, subtract readings at 540 nm or 570 nm from the readings at 450 nm. This subtraction will correct for optical imperfections in the plate. Readings made directly at 450 nm without correction may be higher and less accurate.

Citations for Human Angiopoietin-like 4 DuoSet ELISA

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.

22 Citations: Showing 1 - 10
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  1. Angiopoietin-like 4 promotes osteosarcoma cell proliferation and migration and stimulates osteoclastogenesis
    Authors: T Zhang, A Kastrenopo, Q Larrouture, NA Athanasou, HJ Knowles
    BMC Cancer, 2018;18(1):536.
    Species: Human
    Sample Types: Cell Culture Supernates
  2. Genetic inactivation of ANGPTL4 improves glucose homeostasis and is associated with reduced risk of diabetes
    Authors: V Gusarova, C O'Dushlain, TM Teslovich, PN Benotti, T Mirshahi, O Gottesman, CV Van Hout, MF Murray, A Mahajan, JB Nielsen, L Fritsche, AB Wulff, DF Gudbjartss, M Sjögren, CA Emdin, RA Scott, WJ Lee, A Small, LC Kwee, OP Dwivedi, RB Prasad, S Bruse, AE Lopez, J Penn, A Marcketta, JB Leader, CD Still, HL Kirchner, UL Mirshahi, AH Wardeh, CM Hartle, L Habegger, SN Fetterolf, T Tusie-Luna, AP Morris, H Holm, V Steinthors, P Sulem, U Thorsteins, JI Rotter, LM Chuang, S Damrauer, D Birtwell, CM Brummett, AV Khera, P Natarajan, M Orho-Melan, J Flannick, LA Lotta, CJ Willer, OL Holmen, MD Ritchie, DH Ledbetter, AJ Murphy, IB Borecki, JG Reid, JD Overton, O Hansson, L Groop, SH Shah, WE Kraus, DJ Rader, YI Chen, K Hveem, NJ Wareham, S Kathiresan, O Melander, K Stefansson, BG Nordestgaa, A Tybjærg-Ha, GR Abecasis, D Altshuler, JC Florez, M Boehnke, MI McCarthy, GD Yancopoulo, DJ Carey, AR Shuldiner, A Baras, FE Dewey, J Gromada
    Nat Commun, 2018;9(1):2252.
    Species: Human
    Sample Types: Plasma
  3. Induction of paracrine signaling in metastatic melanoma cells by PPAR? agonist rosiglitazone activates stromal cells and enhances tumor growth
    Authors: C Pich, P Meylan, B Mastelic-G, NT Nguyen, R Loyon, BK Trang, H Moser, C Moret, C Goepfert, J Hafner, MP Levesque, P Romero, C Jandus, L Michalik
    Cancer Res., 2018;0(0):.
    Species: Human
    Sample Types: Cell Culture Supernates
  4. High Concentrations of Angiopoietin-Like Protein 4 Detected in Serum from Patients with Rheumatoid Arthritis Can Be Explained by Non-Specific Antibody Reactivity
    Authors: E Makoveichu, T Ruge, S Nilsson, A S”dergren, G Olivecrona
    PLoS ONE, 2017;12(1):e0168922.
    Species: Human
    Sample Types: Serum
  5. Angptl4 does not control hyperglucagonemia or �-cell hyperplasia following glucagon receptor inhibition
    Authors: H Okamoto, K Cavino, E Na, E Krumm, S Kim, PE Stevis, J Harp, AJ Murphy, GD Yancopoulo, J Gromada
    Proc. Natl. Acad. Sci. U.S.A, 2017;0(0):.
    Species: Human
    Sample Types: Plasma
  6. MicroRNA and mRNA Transcriptome Profiling in Primary Human Astrocytes Infected with Borrelia burgdorferi
    Authors: T Casselli, H Qureshi, E Peterson, D Perley, E Blake, B Jokinen, A Abbas, S Nechaev, JA Watt, A Dhasarathy, CA Brissette
    PLoS ONE, 2017;12(1):e0170961.
    Species: Human
    Sample Types: Cell Culture Supernates
  7. Expression of the angiogenic mediator, angiopoietin-like 4, in the eyes of patients with proliferative sickle retinopathy
    Authors: K Jee, M Rodrigues, F Kashiwabuc, BP Applewhite, I Han, G Lutty, MF Goldberg, GL Semenza, S Montaner, A Sodhi
    PLoS ONE, 2017;12(8):e0183320.
    Species: Human
    Sample Types: Aqueous and Vitreous Humor
  8. Hypoxia-Inducible Factor-Dependent Expression of Angiopoietin-Like 4 by Conjunctival Epithelial Cells Promotes the Angiogenic Phenotype of Pterygia
    Authors: Q Meng, Y Qin, M Deshpande, F Kashiwabuc, M Rodrigues, Q Lu, H Ren, JH Elisseeff, GL Semenza, SV Montaner, A Sodhi
    Invest. Ophthalmol. Vis. Sci., 2017;58(11):4514-4523.
    Species: Human
    Sample Types: Cell Culture Supernates
  9. Angiopoietin-like protein 3 and 4 expression 4 and their serum levels in hepatocellular carcinoma
    Authors: AS El-Shal, HE Zidan, NM Rashad, FM Wadea
    Cytokine, 2017;96(0):75-86.
    Species: Human
    Sample Types: Serum
  10. Angiopoietin-like protein 4 and cardiovascular function in COPD
    Authors: S Janciauski
    BMJ Open Respir Res, 2016;3(1):e000161.
    Species: Human
    Sample Types: Serum
  11. alpha1-Antitrypsin Combines with Plasma Fatty Acids and Induces Angiopoietin-like Protein 4 Expression.
    Authors: Frenzel E, Wrenger S, Brugger B, Salipalli S, Immenschuh S, Aggarwal N, Lichtinghagen R, Mahadeva R, Marcondes A, Dinarello C, Welte T, Janciauskiene S
    J Immunol, 2015;195(8):3605-16.
    Species: Human
    Sample Types: Cell Culture Supernates
  12. Involvement of angiopoietin-like 4 in matrix remodeling during chondrogenic differentiation of mesenchymal stem cells.
    Authors: Mathieu M, Iampietro M, Chuchana P, Guerit D, Djouad F, Noel D, Jorgensen C
    J Biol Chem, 2014;289(12):8402-12.
    Species: Human
    Sample Types: Cell Culture Supernates
  13. Host biomarkers distinguish dengue from leptospirosis in Colombia: a case-control study.
    Authors: Conroy A, Gelvez M, Hawkes M, Rajwans N, Liles W, Villar-Centeno L, Kain K
    BMC Infect Dis, 2014;14(0):35.
  14. Cyclic strain alters the expression and release of angiogenic factors by human tendon cells.
    Authors: Mousavizadeh R, Khosravi S, Behzad H, McCormack R, Duronio V, Scott A
    PLoS ONE, 2014;9(5):e97356.
    Species: Human
    Sample Types: Cell Culture Supernates
  15. Circulating angiopoietin-like 4 links proteinuria with hypertriglyceridemia in nephrotic syndrome.
    Authors: Clement L, Mace C, Avila-Casado C, Joles J, Kersten S, Chugh S
    Nat Med, 2014;20(1):37-46.
    Species: Human
    Sample Types: Plasma
  16. Acute-phase protein alpha1-antitrypsin--a novel regulator of angiopoietin-like protein 4 transcription and secretion.
    Authors: Frenzel E, Wrenger S, Immenschuh S, Koczulla R, Mahadeva R, Deeg H, Dinarello C, Welte T, Marcondes A, Janciauskiene S
    J Immunol, 2014;192(11):5354-62.
  17. The cancer gene WWOX behaves as an inhibitor of SMAD3 transcriptional activity via direct binding.
    Authors: Ferguson B, Gao X, Zelazowski M, Lee J, Jeter C, Abba M, Aldaz C
    BMC Cancer, 2013;13(0):593.
    Species: Human
    Sample Types: Cell Culture Supernates
  18. Epidermal growth factor receptor variant type III markedly accelerates angiogenesis and tumor growth via inducing c-myc mediated angiopoietin-like 4 expression in malignant glioma.
    Authors: Katanasaka Y, Kodera Y, Kitamura Y, Morimoto T, Tamura T, Koizumi F
    Mol Cancer, 2013;12(0):31.
    Species: Human
    Sample Types: Cell Culture Supernates
  19. Disrupting galectin-1 interactions with N-glycans suppresses hypoxia-driven angiogenesis and tumorigenesis in Kaposi's sarcoma.
    J. Exp. Med., 2012;209(11):1985-2000.
    Species: Human
    Sample Types: Cell Culture Supernates
  20. Inactivation of lipoprotein lipase occurs on the surface of THP-1 macrophages where oligomers of angiopoietin-like protein 4 are formed.
    Authors: Makoveichuk E, Sukonina V, Kroupa O, Thulin P, Ehrenborg E, Olivecrona T, Olivecrona G
    Biochem Biophys Res Commun, 2012;425(2):138-43.
    Species: Human
    Sample Types: Cell Culture Supernates
  21. Hypoxia-inducible factor regulates osteoclast-mediated bone resorption: role of angiopoietin-like 4.
    Authors: Knowles HJ, Cleton-Jansen AM, Korsching E, Athanasou NA
    FASEB J., 2010;24(12):4648-59.
    Species: Human
    Sample Types: Cell Culture Supernates
  22. Viral G protein-coupled receptor up-regulates Angiopoietin-like 4 promoting angiogenesis and vascular permeability in Kaposi's sarcoma.
    Authors: Ma T, Jham BC, Hu J
    Proc. Natl. Acad. Sci. U.S.A., 2010;107(32):14363-8.
    Species: Human
    Sample Types: Cell Culture Supernates


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