Human Phospho-Mer DuoSet IC ELISA
R&D Systems, part of Bio-Techne | Catalog # DYC2579-2
Key Product Details
Assay Type
Solid Phase Sandwich ELISA
Sample Type
Cell Lysates
Reactivity
Human
Human Phospho-Mer DuoSet IC ELISA 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
Product Summary for Human Phospho-Mer DuoSet IC ELISA
This DuoSet IC ELISA contains the basic components required for the development of sandwich ELISAs to measure tyrosine-phosphorylated human Mer in cell lysates. An immobilized capture antibody specific for Mer binds both phosphorylated and unphosphorylated Mer . After washing away unbound material, a biotinylated detection antibody is used to detect only phosphorylated receptor, utilizing a standard 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 Phospho-Mer DuoSet IC ELISA
The Human Phospho-Mer DuoSet IC ELISA is more sensitive than immunoprecipitation (IP)-Western analysis
NS0 cells transfected with human Mer (NS0-hMer) were treated with conditioned media from the human lung cell line, WI-38, for 15 minutes to induce tyrosine phosphorylation of Mer. Lysates were serially diluted and analyzed by (A) IP-Western blot and (B) this DuoSet IC ELISA. IPs were done using an anti-Mer monoclonal antibody and goat anti-mouse agarose. Immunoblots were incubated with an HRP-conjugated anti-phosphotyrosine monoclonal antibody (R&D Systems, Catalog # HAM1676) to detect phospho-Mer. Bands were visualized by chemiluminescent detection using WesternGloTM Chemiluminescent Detection Substrate (R&D Systems, Catalog # AR004). Human Phospho-Mer can be detected in this DuoSet IC ELISA by using approximately 5 to 10 times less lysate than is needed for a conventional IP-Western blot.
The Human Phospho-MerDuoSet IC ELISA detects ligand-induced Mer tyrosine phosphorylation
NS0-hMer cells were untreated or treated with conditioned media from the human lung cancer cell line, WI-38, for 15 minutes. ELISA and IP-Western blot (inset) analyses were done using 100 μg and 400 μg of lysate, respectively. IP-Western blots for phospho-Mer (p-Mer) were done as described in Figure 1. Blots were stripped and total Mer was detected using a biotinylated anti-Mer polyclonal antibody (R&D Systems, Catalog #BAF891).
The specificity of the Human Phospho-MerDuoSet IC ELISA is confirmed by receptor competition
NS0-hMer cells were treated with conditioned media from the human lung cell line, WI-38, for 15 minutes minutes. The indicated amounts of recombinant extracellular domains of human Mer/Fc (R&D Systems, Catalog #891-MR), human Axl/Fc (R&D Systems, Catalog #154-AL), human Dtk/Fc (R&D Systems, Catalog #859-DK) or human TrkA/Fc (R&D Systems, Catalog #175-TK) were added to 100 μg lysate and analyzed using this DuoSet IC ELISA. Competition was observed only with recombinant human Mer.Kit Contents for Human Phospho-Mer DuoSet IC ELISA
- 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
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: Mer
Long Name
Receptor Tyrosine Protein Kinase Mer
Alternate Names
c-Eyk, C-mer, MerTK
Gene Symbol
MERTK
Additional Mer Products
Product Documents for Human Phospho-Mer DuoSet IC ELISA
Product Specific Notices for Human Phospho-Mer DuoSet IC ELISA
For research use only
Loading...
Loading...
Loading...
Loading...