Human Total Axl DuoSet IC ELISA

R&D Systems | Catalog # DYC1643-2

R&D Systems
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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
View all Axl ELISA Kits »

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

Please see the product datasheet

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.

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.

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

Entrez Gene IDs

558 (Human); 26362 (Mouse); 102126361 (Cynomolgus Monkey)

Gene Symbol

AXL

Additional Axl Products

Product Documents for Human Total Axl DuoSet IC ELISA

Certificate of Analysis

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Product Specific Notices for Human Total Axl DuoSet IC ELISA

For research use only

Citations for Human Total Axl DuoSet IC ELISA

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