Human Axl DuoSet ELISA

Catalog # Availability Size / Price Qty
DY154
Ancillary Products Available
Human Axl ELISA Standard Curve
1 Image
Product Details
Procedure
Citations (14)
FAQs
Supplemental Products
Reviews (2)

Human Axl DuoSet ELISA Summary

Assay Type
Solid Phase Sandwich ELISA
Format
96-well strip plate
Sample Volume Required
100 µL
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 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)

Data Example

Human Axl ELISA Standard Curve

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: 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.

Long Name:
Axl Receptor Tyrosine Kinase
Entrez Gene IDs:
558 (Human); 26362 (Mouse)
Alternate Names:
AI323647; ARK; AXL oncogene; AXL receptor tyrosine kinase; AXL transforming sequence/gene; Axl; EC 2.7.10; EC 2.7.10.1; JTK11; Tyro7; tyrosine-protein kinase receptor UFO; UFO; UFOoncogene AXL

Assay Procedure

GENERAL ELISA PROTOCOL

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 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.

14 Citations: Showing 1 - 10
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  1. 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;2020(0):2696173.
    Species: Human
    Sample Types: Plasma
  2. 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;15(1):e0227187.
    Species: Human
    Sample Types: Serum
  3. 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;12(4):.
    Species: Mouse
    Sample Types: Plasma
  4. 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;0(0):.
    Species: Human
    Sample Types: Serum
  5. 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;14(2):e0212068.
    Species: Human
    Sample Types: Serum
  6. Soluble AXL is ubiquitously present in malignant serous effusions
    Authors: K Flem Karls, E McFadden, VA Flørenes, B Davidson
    Gynecol. Oncol., 2018;0(0):.
    Species: Human
    Sample Types: Cell Culture Supernates
  7. 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;13(4):e0195724.
    Species: Human
    Sample Types: Plasma
  8. 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;8(0):15877.
    Species: Human
    Sample Types: Serum
  9. 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;8(28):46234-46248.
    Species: Human
    Sample Types: Serum
  10. 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;68(9):1303-9.
    Species: Human
    Sample Types: Serum
  11. ADAM17 substrate release in proximal tubule drives kidney fibrosis
    JCI Insight, 2016;1(13):.
    Species: Human
    Sample Types: Urine
  12. 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;9(12):e115916.
    Species: Human
    Sample Types: Plasma
  13. 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;4(11):e7916.
    Species: Human
    Sample Types: Plasma
  14. 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;183(5):3004-13.
    Species: Human
    Sample Types: Cell Culture Supernates

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Human Axl DuoSet ELISA
By Anonymous on 07/11/2018
Application: Sample Tested: EDTA Plasma

Human Axl DuoSet ELISA
By Anonymous on 07/09/2018
Application: Sample Tested: Recombinant protein