Human Gas6 DuoSet ELISA Summary
* 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 Growth Arrest-Specific Protein 6 (Gas6). The suggested diluent is suitable for the analysis of most cell culture supernate, serum, plasma, and urine samples. Diluents for complex matrices, such as serum, plasma, and urine, 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)
Scientific Data
Product Datasheets
Preparation and Storage
Background: Gas6
Gas6 (Growth Arrest Specific 6) is a secreted protein that contains an extensively gamma-carboxylated N-terminal Gla domain and is dependent upon vitamin K for activity. Gas6 signals through the receptors Axl, Dtk/Tyro3, and Mer and is inhibited by shed soluble forms of Axl and Mer. Gas6 protects cells from stress-induced apoptosis and promotes phagocytosis of apoptotic cell debris, promotes platelet activation but inhibits inflammatory cytokine production, and inhibits VEGF-induced angiogenesis, and regulates tumor cell invasiveness.
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 Gas6 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.
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Citations: Showing 1 - 10
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Down-regulated GAS6 impairs synovial macrophage efferocytosis and promotes obesity-associated osteoarthritis
Authors: Yao, Z;Qi, W;Zhang, H;Zhang, Z;Liu, L;Shao, Y;Zeng, H;Yin, J;Pan, H;Guo, X;Liu, A;Cai, D;Bai, X;Zhang, H;
eLife
Species: Human
Sample Types: Synovial Fluid
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Vitamin K Effects on Gas6 and Soluble Axl Receptors in Intensive Care Patients: An Observational Screening Study
Authors: U Schött, C Augustsson, L Lilover, CU Nilsson, L Walther-St, T Kander
Nutrients, 2021-11-16;13(11):.
Species: Human
Sample Types: Plasma
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Evolution of NETosis markers and DAMPs have prognostic value in critically ill COVID-19 patients
Authors: J Huckriede, SB Anderberg, A Morales, F de Vries, M Hultström, A Bergqvist, JT Ortiz-Pére, JW Sels, K Wichapong, M Lipcsey, M van de Pol, A Larsson, T Luther, C Reutelings, PG de Frutos, R Frithiof, GAF Nicolaes
Scientific Reports, 2021-08-03;11(1):15701.
Species: Human
Sample Types: Plasma
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Gamma-glutamyl carboxylated Gas6 facilitates the prophylactic effect of vitamin K in inhibiting hyperlipidemia-associated inflammatory pathophysiology via arresting MCP-1/ICAM-1 mediated monocyte-hepatocyte adhesion
Authors: J Bordoloi, D Ozah, T Bora, J Kalita, P Manna
The Journal of nutritional biochemistry, 2021-03-28;0(0):108635.
Species: Human
Sample Types: Plasma
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Analysis of Inflammatory Mediator Profiles in Sepsis Patients Reveals That Extracellular Histones Are Strongly Elevated in Nonsurvivors
Authors: T Eichhorn, I Linsberger, L Lauková, C Tripiscian, B Fendl, R Weiss, F König, G Valicek, G Miestinger, C Hörmann, V Weber
Mediators of Inflammation, 2021-03-17;2021(0):8395048.
Species: Human
Sample Types: Plasma
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Selective Increment of Synovial Soluble TYRO3 Correlates with Disease Severity and Joint Inflammation in Patients with Rheumatoid Arthritis
Authors: J Vullings, JP Vago, CEJ Waterborg, RM Thurlings, MI Koenders, PLEM van Lent, PM van der Kr, FA Amaral, FAJ van de Loo
J Immunol Res, 2020-09-11;2020(0):9690832.
Species: Human
Sample Types: Synovial Fluid
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AXL confers cell migration and invasion by hijacking a PEAK1-regulated focal adhesion protein network
Authors: A Abu-Thurai, MA Goyette, J Boulais, C Delliaux, C Apcher, C Schott, R Chidiac, H Bagci, MP Thibault, D Davidson, M Ferron, A Veillette, RJ Daly, AC Gingras, JP Gratton, JF Côté
Nat Commun, 2020-07-17;11(1):3586.
Species: Human
Sample Types: Cell Culture Supernates
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Distinct synovial tissue macrophage subsets regulate inflammation and remission in rheumatoid arthritis
Authors: S Alivernini, L MacDonald, A Elmesmari, S Finlay, B Tolusso, MR Gigante, L Petricca, C Di Mario, L Bui, S Perniola, M Attar, M Gessi, AL Fedele, S Chilaka, D Somma, SN Sansom, A Filer, C McSharry, NL Millar, K Kirschner, A Nerviani, MJ Lewis, C Pitzalis, AR Clark, G Ferracciol, I Udalova, CD Buckley, E Gremese, IB McInnes, TD Otto, M Kurowska-S
Nat. Med., 2020-06-29;0(0):.
Species: Human
Sample Types: Cell Culture Supernates
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Respiratory syncytial virus infection exacerbates pneumococcal pneumonia via Gas6/Axl-mediated macrophage polarization
Authors: T Shibata, A Makino, R Ogata, S Nakamura, T Ito, K Nagata, Y Terauchi, T Oishi, M Fujieda, Y Takahashi, M Ato
J. Clin. Invest., 2020-06-01;0(0):.
Species: Human
Sample Types: Cell Culture Superantes
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Efficient blockade of locally reciprocated tumor-macrophage signaling using a TAM-avid nanotherapy
Authors: SJ Wang, R Li, TSC Ng, G Luthria, MJ Oudin, M Prytyskach, RH Kohler, R Weissleder, DA Lauffenbur, MA Miller
Sci Adv, 2020-05-22;6(21):eaaz8521.
Species: Human
Sample Types: Cell Lysates
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Hematopoietic niche drives FLT3-ITD acute myeloid leukemia resistance to quizartinib via STAT5- and hypoxia- dependent up-regulation of AXL
Authors: PY Dumas, C Naudin, S Martin-Lan, B Izac, L Casetti, O Mansier, B Rousseau, A Artus, M Dufossée, A Giese, P Dubus, A Pigneux, V Praloran, A Bidet, A Villacrece, AV Guitart, N Milpied, O Kosmider, I Vigon, V Desplat, I Dusanter-F, JM Pasquet
Haematologica, 2019-03-28;0(0):.
Species: Human
Sample Types: Cell Culture Supernates
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Expression and role of TYRO3 and AXL as potential therapeutical targets in leiomyosarcoma
Authors: C Dantas-Bar, T Lesluyes, FL Loarer, F Chibon, I Treilleux, JM Coindre, P Meeus, M Brahmi, O Bally, I Ray-Coquar, MP Sunyach, AL Cesne, O Mir, S Bonvalot, M Toulmonde, A Italiano, P Saintigny, M Jean-Denis, F Ducimetier, D Ranchere, H El Sayadi, L Alberti, JY Blay
Br. J. Cancer, 2017-10-12;117(12):1787-1797.
Species: Human
Sample Types: Cell Lysates
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Viral Infection Sensitizes Human Fetal Membranes to Bacterial Lipopolysaccharide by MERTK Inhibition and Inflammasome Activation
Authors: SN Cross, JA Potter, P Aldo, JY Kwon, M Pitruzzell, M Tong, S Guller, CV Rothlin, G Mor, VM Abrahams
J. Immunol., 2017-09-15;0(0):.
Species: Human
Sample Types: Tissue Homogenates
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MicroRNA-34a dependent regulation of AXL controls the activation of dendritic cells in inflammatory arthritis
Authors: M Kurowska-S, S Alivernini, EG Melchor, A Elmesmari, B Tolusso, C Tange, L Petricca, DS Gilchrist, G Di Sante, C Keijzer, L Stewart, C Di Mario, V Morrison, JM Brewer, D Porter, S Milling, RD Baxter, D McCarey, E Gremese, G Lemke, G Ferracciol, C McSharry, IB McInnes
Nat Commun, 2017-06-22;8(0):15877.
Species: Human
Sample Types: Serum
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Axl is required for TGF-?2-induced dormancy of prostate cancer cells in the bone marrow
Sci Rep, 2016-11-07;6(0):36520.
Species: Human
Sample Types: Cell Lysates
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Role of Growth arrest-specific gene 6-Mer axis in multiple myeloma.
Authors: Waizenegger J, Ben-Batalla I, Weinhold N, Meissner T, Wroblewski M, Janning M, Riecken K, Binder M, Atanackovic D, Taipaleenmaeki H, Schewe D, Sawall S, Gensch V, Cubas-Cordova M, Seckinger A, Fiedler W, Hesse E, Kroger N, Fehse B, Hose D, Klein B, Raab M, Pantel K, Bokemeyer C, Loges S
Leukemia, 2014-08-08;29(3):696-704.
Species: Human
Sample Types: Bone Marrow Fluid
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Plasma concentrations predict aortic expression of growth-arrest-specific protein 6 in patients undergoing coronary artery bypass grafting.
Authors: Lee, Chien-Hs, Shieh, Yi-Shing, Tsai, Chien-Su, Hung, Yi-Jen, Tsai, Yi-Ting, Lin, Chih-Yua
PLoS ONE, 2013-11-13;8(11):e79452.
Species: Human
Sample Types: Plasma
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GAS6/Mer axis regulates the homing and survival of the E2A/PBX1-positive B-cell precursor acute lymphoblastic leukemia in the bone marrow niche.
Authors: Shiozawa Y, Pedersen EA, Taichman RS
Exp. Hematol., 2009-11-14;38(2):132-40.
Species: Human
Sample Types: Cell Culture Supernates
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