Human Phospho-Axl DuoSet IC ELISA

Catalog # Availability Size / Price Qty
DYC2228E
DYC2228-5
DYC2228-2
Human Phospho-Axl DuoSet IC ELISA Data
3 Images
Product Details
Citations (3)
FAQs
Supplemental Products
Reviews

Human Phospho-Axl DuoSet IC ELISA Summary

Assay Type
Solid Phase Sandwich ELISA
Format
96-well strip plate
Sample Volume Required
Cell lysates (100 µL)
Sufficient Materials
Kits available for two, five, or 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 IC ELISA contains the basic components required for the development of sandwich ELISAs to measure tyrosine-phosphorylated human Axl in cell lysates. An immobilized capture antibody specific for Axl binds both phosphorylated and unphosphorylated Axl. After washing away unbound material, an HRP-conjugated detection antibody is used to detect only phosphorylated receptor, utilizing a standard HRP format.

Product Features

  • Optimized capture and detection antibody pairings and recommended concentrations save lengthy development time
  • Development protocols are provided to guide further assay optimization
  • Assay can be customized to your specific needs
  • Available in 2, 5, and 15-(96-well) plate pack sizes
  • Economical alternative to Western blot

Kit Content

  • Capture Antibody
  • Conjugated Detection Antibody
  • Calibrated Immunoassay Standard or Control

Other Reagents Required


PBS: (Catalog # DY006), or 137 mM NaCl, 2.7 mM KCl, 8.1 mM Na2HPO4, 1.5 mM KH2O4, pH 7.2 - 7.4, 0.2 µm filtered

Wash Buffer: (Catalog # WA126), or equivalent

Lysis Buffer*

IC Diluent*

Blocking Buffer*


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), or equivalent

Plate Sealers: ELISA Plate Sealers (Catalog # DY992), or equivalent

*For the Lysis Buffer, IC Diluent, and Blocking BUffer recommended for a specific DuoSet ELISA Development Kit, please see the product

Data Examples

Figure 1. The Human Phospho-Axl DuoSet IC ELISA is more sensitive than immunoprecipitation (IP)-Western analysis Human glioblastoma cell line, A172, was treated with conditioned media from the human lung cell line, WI38, for fifteen minutes to induce tyrosine phosphorylation of Axl. Serial dilutions of lysates were analyzed by (A) IP-Western blot and (B) this ELISA. IPs were done using an anti-Axl monoclonal antibody and anti-mouse IgG agarose. Immunoblots were incubated with a biotinylated anti-phosphotyrosine monoclonal antibody (Catalog # BAM1676) to detect phospho-Axl (p-Axl). Bands were visualized with Streptavidin-HRP (Cat # DY998) followed by chemiluminescent detection.

Figure 2. The Human Phospho-AxlDuoSet IC ELISA detects ligand-induced Axl tyrosine phosphorylation A172 cells were untreated or treated with conditioned media from the human lung cell line, WI38, for fifteen minutes. ELISA and IP-Western blot (inset) analyses were done using 100 μg and 400 μg of lysate, respectively. IP-Western blots for phospho-Axl (p-Axl) were done as described in Figure 1. Blots were stripped and total Axl was detected using a biotinylated anti-Axl polyclonal antibody (Catalog # BAF154).

Figure 3. The specificity of the Human Phospho-AxlDuoSet IC ELISA is confirmed by receptor competition A172 cells were treated with conditioned media from the human lung cell line WI38 for fifteen minutes. The indicated amounts of recombinant extracellular domains of human Axl (Catalog # 154-AL), human Dtk (Catalog # 859-DK) or human Trk A (Catalog # 175-TK) were added to 125 μg lysate and analyzed using this ELISA. Competition was observed only with recombinant Axl.

Product Datasheets

Preparation and Storage

Shipping
The product is shipped with polar packs. 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

Citations for Human Phospho-Axl DuoSet IC 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.

3 Citations: Showing 1 - 3
Filter your results:

Filter by:

  1. Increased secretion of Gas6 by smooth muscle cells in human atherosclerotic carotid plaques.
    Authors: Clauser S, Meilhac O, Bieche I, Raynal P, Bruneval P, Michel JB, Borgel D
    Thromb. Haemost., 2011;107(1):140-9.
    Species: Human
    Sample Types: Cell Lysates
  2. Human cutaneous melanomas lacking MITF and melanocyte differentiation antigens express a functional Axl receptor kinase.
    Authors: Sensi M, Catani M, Castellano G
    J. Invest. Dermatol., 2011;131(12):2448-57.
    Species: Human
    Sample Types: Cell Lysates
  3. An anti-Axl monoclonal antibody attenuates xenograft tumor growth and enhances the effect of multiple anticancer therapies.
    Authors: Ye X, Li Y, Stawicki S
    Oncogene, 2010;29(38):5254-64.
    Species: Human
    Sample Types: Cell Lysates

FAQs

No product specific FAQs exist for this product, however you may

View all ELISA FAQs

Reviews for Human Phospho-Axl DuoSet IC ELISA

There are currently no reviews for this product. Be the first to review Human Phospho-Axl DuoSet IC ELISA and earn rewards!

Have you used Human Phospho-Axl DuoSet IC ELISA?

Submit a review and receive an Amazon gift card.

$25/€18/£15/$25CAN/¥75 Yuan/¥1250 Yen for a review with an image

$10/€7/£6/$10 CAD/¥70 Yuan/¥1110 Yen for a review without an image

Submit a Review