Human Angiopoietin-like 4 DuoSet ELISA
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Summary
Product Features
Kit Content
Other Reagents Required
Data Examples
Product Datasheets
Preparation and Storage
Background
Human Angiopoietin-like 4 DuoSet ELISA Summary
Assay Type
Solid Phase Sandwich ELISA
Format
96-well strip plate
Sample Volume Required
100 µL
Assay Range
1.3 - 80 ng/mL
Sufficient Materials
For fifteen 96-well plates*
Specificity
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.
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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)
Scientific Data
Product Datasheets
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 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
GENERAL ELISA PROTOCOL
Plate Preparation
- 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.
- 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.
- Block plates by adding 300 μL Reagent Diluent to each well. Incubate at room temperature for a minimum of 1 hour.
- Repeat the aspiration/wash as in step 2. The plates are now ready for sample addition.
Assay Procedure
- 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.
- Repeat the aspiration/wash as in step 2 of Plate Preparation.
- 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.
- Repeat the aspiration/wash as in step 2 of Plate Preparation.
- 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.
- Repeat the aspiration/wash as in step 2.
- Add 100 μL of Substrate Solution to each well. Incubate for 20 minutes at room temperature. Avoid placing the plate in direct light.
- Add 50 μL of Stop Solution to each well. Gently tap the plate to ensure thorough mixing.
- 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.
31
Citations: Showing 1 - 10
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ANGPTL4, IL-6 and TNF-alpha as regulators of lipid metabolism during a marathon run
Authors: M Górecka, K Krzemi?ski, T Mikulski, AW Ziemba
Scientific Reports, 2022;12(1):19940.
Species: Human
Sample Types: Plasma
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GALNT2 rs4846914 SNP Is Associated with Obesity, Atherogenic Lipid Traits, and ANGPTL3 Plasma Level
Authors: M Qaddoumi, P Hebbar, M Abu-Farha, A Al Somaly, M Melhem, F Al-Kayal, I AlKhairi, P Cherian, M Alanbaei, F Al-Mulla, J Abubaker, TA Thanaraj
Genes, 2022;13(7):.
Species: Human
Sample Types: Plasma
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ANGPTL4 influences the therapeutic response of neovascular age-related macular degeneration patients by promoting choroidal neovascularization
Authors: Y Qin, A Dinabandhu, X Cao, JC Sanchez, K Jee, M Rodrigues, C Guo, J Zhang, J Vancel, D Menon, NS Khan, T Ma, SY Tzeng, YJ Daoud, JJ Green, GL Semenza, S Montaner, A Sodhi
JCI Insight, 2022;0(0):.
Species: Human
Sample Types: Aqueous Humor
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ANGPTL4 Expression Is Increased in Epicardial Adipose Tissue of Patients with Coronary Artery Disease
Authors: Y Katanasaka, A Saito, Y Sunagawa, N Sari, M Funamoto, S Shimizu, K Shimizu, T Akimoto, C Ueki, M Kitano, K Hasegawa, G Sakaguchi, T Morimoto
Journal of Clinical Medicine, 2022;11(9):.
Species: Human
Sample Types: Pericardial Fluid
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Circulating ANGPTL3 and ANGPTL4 levels predict coronary artery atherosclerosis severity
Authors: T Sun, W Zhan, L Wei, Z Xu, L Fan, Y Zhuo, C Wang, J Zhang
Lipids in Health and Disease, 2021;20(1):154.
Species: Human
Sample Types: Plasma
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Decreased ANGPTL4 impairs endometrial angiogenesis during peri-implantation period in patients with recurrent implantation failure
Authors: M Li, J Hu, L Yao, M Gao
J. Cell. Mol. Med., 2020;0(0):.
Species: Human
Sample Types: Serum
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Fasting induces ANGPTL4 and reduces LPL activity in human adipose tissue
Authors: PMM Ruppert, CCJR Michielsen, EJ Hazebroek, A Pirayesh, G Olivecrona, LA Afman, S Kersten
Mol Metab, 2020;40(0):101033.
Species: Human
Sample Types: Plasma
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Decreased ANGPTL4 impairs endometrial angiogenesis during peri-implantation period in patients with recurrent implantation failure
Authors: M Li, J Hu, L Yao, M Gao
J. Cell. Mol. Med., 2020;0(0):.
Species: Human
Sample Types: Serum
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Hypocholesterolemia and dysregulated production of angiopoietin-like proteins in sickle cell anemia patients
Authors: F Vendrame, L Olops, STO Saad, FF Costa, KY Fertrin
Cytokine, 2019;120(0):88-91.
Species: Human
Sample Types: Serum
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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
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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
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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
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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
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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
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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
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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
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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
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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
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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
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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
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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
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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.
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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.
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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
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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
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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
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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
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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
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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
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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
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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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