Human Axl 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 Axl. 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)
Scientific Data
Product Datasheets
Preparation and Storage
Background: Axl
Axl (Ufo, Ark), Dtk (Sky, Tyro3, Rse, Brt) and Mer (human and mouse orthologs of chicken c-Eyk) constitute the TAM receptor tyrosine kinase subfamily. This RTK subfamily is characterized by an extracellular domain that consists of two Ig-like motifs and two fibronectin type III motifs. These receptors bind the vitamin K-dependent protein Growth Arrest Specific Gene 6 (Gas6). Receptor activation leads to cell proliferation, migration, or the prevention of apoptosis. Cellular signaling through this family of RTKs is involved in hematopoiesis, embryonic development, tumorigenesis, and spermatogenesis.
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 Axl 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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Evaluating soluble Axl as a biomarker for glioblastoma: A pilot study
Authors: Raymond, D;Fukui, M;Zwernik, S;Kassam, A;Rovin, R;Akhtar, P;
PloS one
Species: Human
Sample Types: Serum, Cell Culture Supernates
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Expression and clinical association of MFG-E8 and TAM receptors in diabetic patients with different stages of microvascular complication: An experimental study
Authors: Xiong, Y;Wang, B;Jiao, J;Tang, M;Li, K;Jiang, R;Huang, W;
Medicine
Species: Human
Sample Types: Serum
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Transcriptomic analysis reveals partial epithelial-mesenchymal transition and inflammation as common pathogenic mechanisms in hypertensive nephrosclerosis and Type 2 diabetic nephropathy
Authors: Nordbø, OP;Landolt, L;Eikrem, Ø;Scherer, A;Leh, S;Furriol, J;Apeland, T;Mydel, P;Marti, HP;
Physiological reports
Species: Human
Sample Types: Serum
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Decreased Gas6 and sAxl Plasma Levels Are Associated with Hair Loss in COVID-19 Survivors
Authors: D Apostolo, D D'Onghia, S Tonello, R Minisini, A Baricich, C Gramaglia, F Patrucco, P Zeppegno, A Acquaviva, PE Balbo, LM Castello, G Cappellano, A Chiocchett, C Gerevini, M Giordano, F Laaguid, M Manfredi, D Raineri, C Rigamonti, R Rolla, V Romano, M Confalonie, P Savoia, E Zavattaro, M Pirisi, B Ruaro, PP Sainaghi, M Bellan, No-More CO
International Journal of Molecular Sciences, 2023-03-26;24(7):.
Species: Human
Sample Types: Plasma
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Baseline Plasma Gas6 Protein Elevation Predicts Adverse Outcomes in Hospitalized COVID-19 Patients
Authors: S Tonello, M Rizzi, E Matino, M Costanzo, GF Casciaro, A Croce, E Rizzi, E Zecca, A Pedrinelli, V Vassia, R Landi, M Bellan, LM Castello, R Minisini, VR Mallela, D D'Onghia, GC Avanzi, M Pirisi, D Lilleri, PP Sainaghi
Disease Markers, 2022-04-29;2022(0):1568352.
Species: Human
Sample Types: Plasma
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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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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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Plasma biomarkers of the amyloid pathway are associated with geographic atrophy secondary to age-related macular degeneration
Authors: K Lashkari, GC Teague, U Beattie, J Betts, S Kumar, MM McLaughlin, FJ López
PLoS ONE, 2020-08-07;15(8):e0236283.
Species: Human
Sample Types: Plasma
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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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Role of Gas6 and TAM Receptors in the Identification of Cardiopulmonary Involvement in Systemic Sclerosis and Scleroderma Spectrum Disorders
Authors: M Bellan, A Dimagli, C Piccinino, A Giubertoni, A Ianniello, F Grimoldi, M Sguazzotti, A Nerviani, M Barini, A Carriero, C Smirne, ME Burlone, C Rigamonti, R Minisini, L Salmi, MN Barbaglia, LM Castello, D Sola, P Marino, GC Avanzi, M Pirisi, PP Sainaghi
Dis. Markers, 2020-05-12;2020(0):2696173.
Species: Human
Sample Types: Plasma
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A Novel Treatment Modality for Malignant Peripheral Nerve Sheath Tumor Using a Dual-Effect Liposome to Combine Photodynamic Therapy and Chemotherapy
Authors: CT Chen, PC Peng, T Tsai, HF Chien, MJ Lee
Pharmaceutics, 2020-04-02;12(4):.
Species: Mouse
Sample Types: Plasma
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Soluble AXL as a marker of disease progression and survival in melanoma
Authors: K Flem-Karls, M Nyakas, IN Farstad, E McFadden, P Wernhoff, KD Jacobsen, VA Flørenes, GM Mælandsmo
PLoS ONE, 2020-01-09;15(1):e0227187.
Species: Human
Sample Types: Serum
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A FUNCTIONAL ROLE OF GAS6/TAM IN NON-ALCOHOLIC STEATOHEPATITIS PROGRESSION IMPLICATES AXL AS THERAPEUTIC TARGET
Authors: A Tutusaus, E de Gregori, B Cucarull, H Cristóbal, C Aresté, I Graupera, M Coll, A Colell, G Gausdal, JB Lorens, P García de, A Morales, M Marí
Cell Mol Gastroenterol Hepatol, 2019-11-02;0(0):.
Species: Human
Sample Types: Serum
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Serum Axl predicts histology-based response to induction therapy and long-term renal outcome in lupus nephritis
Authors: I Parodis, H Ding, A Zickert, G Cosson, M Fathima, C Grönwall, C Mohan, I Gunnarsson
PLoS ONE, 2019-02-11;14(2):e0212068.
Species: Human
Sample Types: Serum
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Soluble AXL is ubiquitously present in malignant serous effusions
Authors: K Flem Karls, E McFadden, VA Flørenes, B Davidson
Gynecol. Oncol., 2018-11-15;0(0):.
Species: Human
Sample Types: Cell Culture Supernates
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Imbalance between endothelial damage and repair capacity in chronic obstructive pulmonary disease
Authors: J García-Luc, VI Peinado, L de Jover, R Del Pozo, I Blanco, C Bonjoch, N Coll-Bonfi, T Paul, O Tura-Ceide, JA Barberà
PLoS ONE, 2018-04-19;13(4):e0195724.
Species: Human
Sample Types: Plasma
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Soluble Axl is an accurate biomarker of cirrhosis and hepatocellular carcinoma development: results from a large scale multicenter analysis
Authors: M Dengler, K Staufer, H Huber, R Stauber, H Bantel, KH Weiss, P Starlinger, H Pock, P Klöters-Pl, DN Gotthardt, P Rauch, C Lackner, J Stift, C Brostjan, T Gruenberge, T Kumada, H Toyoda, T Tada, TS Weiss, M Trauner, W Mikulits
Oncotarget, 2017-07-11;8(28):46234-46248.
Species: Human
Sample Types: Serum
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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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ADAM17 substrate release in proximal tubule drives kidney fibrosis
JCI Insight, 2016-08-18;1(13):.
Species: Human
Sample Types: Urine
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Axl, Ferritin, Insulin-Like Growth Factor Binding Protein 2, and Tumor Necrosis Factor Receptor Type II as Biomarkers in Systemic Lupus Erythematosus
Authors: CC Mok, HH Ding, M Kharboutli, C Mohan
Arthritis Care Res (Hoboken), 2016-07-28;68(9):1303-9.
Species: Human
Sample Types: Serum
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Soluble AXL: a possible circulating biomarker for neurofibromatosis type 1 related tumor burden.
Authors: Johansson, Gunnar, Peng, Po-Chun, Huang, Po-Yuan, Chien, Hsiung-F, Hua, Kuo-Tai, Kuo, Min-Lian, Chen, Chin-Tin, Lee, Ming-Jen
PLoS ONE, 2014-12-31;9(12):e115916.
Species: Human
Sample Types: Plasma
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Integrated proteomic analysis of human cancer cells and plasma from tumor bearing mice for ovarian cancer biomarker discovery.
Authors: Pitteri SJ, JeBailey L, Faca VM, Thorpe JD, Silva MA, Ireton RC, Horton MB, Wang H, Pruitt LC, Zhang Q, Cheng KH, Urban N, Hanash SM, Dinulescu DM
PLoS ONE, 2009-11-19;4(11):e7916.
Species: Human
Sample Types: Plasma
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Survival and migration of human dendritic cells are regulated by an IFN-alpha-inducible Axl/Gas6 pathway.
Authors: Scutera S, Fraone T, Musso T, Cappello P, Rossi S, Pierobon D, Orinska Z, Paus R, Bulfone-Paus S, Giovarelli M
J. Immunol., 2009-08-05;183(5):3004-13.
Species: Human
Sample Types: Cell Culture Supernates
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