Human Total Axl DuoSet IC ELISA
R&D Systems, part of Bio-Techne | Catalog # DYC1643-2
Key Product Details
Assay Type
Solid Phase Sandwich ELISA
Assay Range
62.5-4000 pg/mL
Sample Type
Cell Lysates
Reactivity
Human
Human Total Axl DuoSet IC ELISA 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
- Available in 2, 5, and 15- (96-well) plate pack sizes
- Economical alternative to Western blot
Product Summary for Human Total Axl DuoSet IC ELISA
This DuoSet IC ELISA contains the basic components required for the development of sandwich ELISAs to measure total Axl in cell lysates. An immobilized capture antibody specific for Axl binds both phosphorylated and unphosphorylated Axl. After washing away unbound material, a biotinylated detection antibody is used to detect both phosphorylated and unphosphorylated protein, utilizing a standard Streptavidin-HRP format.
Product Specifications
Assay Format
96-well strip plate (sold separately)
Sample Volume Required
Cell lysates (100 µL)
Detection Method
Colorimetric ELISA - 450nm (TMB)
Conjugate
Biotin
Specificity
Label
HRP
Scientific Data Images for Human Total Axl DuoSet IC ELISA
Standard Curve
The Human Total Axl DuoSet® IC ELISA is more sensitive than immunoprecipitation (IP)-Western Blot analysis.
Lysates prepared from the human glioblastoma cell line A172 were diluted in series and analyzed by (A) IP-Western Blot and (B) this DuoSet® IC ELISA. IPs were performed using an anti-Axl monoclonal antibody and goat anti-mouse agarose. Immunoblots were incubated with a Biotinylated Anti-Axl Polyclonal Antibody (Catalog # BAF154) to detect human total Axl. Bands were visualized with Streptavidin-HRP A (Catalog # DY998) followed by chemiluminescent detection. Human Axl can be detected in this DuoSet® IC ELISA by using approximately 10 to 20 times less lysate than is needed for a conventional IP-Western Blot.
The Human Total Axl DuoSet® IC ELISA measures the relative level of Axl.
Lysates were prepared from the human glioblastoma cell line (A172), the human epidermoid carcinoma cell line (A431), and the human stomach cancer cell line (KatoIII). DuoSet® IC ELISA and IP-Western Blot (inset) analyses were performed using 50 μg and 100 μg of lysate, respectively. The IP-Western Blot was performed as described in Figure 1. The DuoSet® IC ELISA results correlate well with the amounts of human Axl detected by IP-Western Blot.Kit Contents for Human Total Axl DuoSet IC ELISA
- Capture Antibody
- Conjugated Detection Antibody
- Calibrated Immunoassay Standard or Control
- Streptavidin-HRP
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: From Costar EIA Plate (Costar Catalog # 2592) or 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
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, also known as Ufo and Ark, is a widely expressed glycoprotein in the TAM receptor tyrosine kinase family. TAM family receptors (Dtk/Tyro3, Axl, and Mer) are involved in regulation of the inflammatory response, cell survival and migration, and tumorigenesis. Axl binds the vitamin K-dependent protein Gas6, triggering platelet aggregation and thrombus formation, macrophage and dendritic cell phagocytosis of apoptotic cells, NK cell development from hematopoietic progenitor cells, and in vivo angiogenesis. Axl is highly expressed in solid cancers and promotes in vivo tumorigenesis and tumor cell invasiveness. It contributes to vascular remodeling and inflammatory cell infiltration in response to hypertension and restricted blood flow. It also functions as a cellular entry receptor for Gas6-opsonized lentiviruses.
Long Name
Axl Receptor Tyrosine Kinase
Alternate Names
AI323647, ARK, JTK11, Tyro7, UFO
Gene Symbol
AXL
Additional Axl Products
Product Documents for Human Total Axl DuoSet IC ELISA
Product Specific Notices for Human Total Axl DuoSet IC ELISA
For research use only
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