Human Total FGFR4 DuoSet IC ELISA
R&D Systems, part of Bio-Techne | Catalog # DYC685-2
Key Product Details
Assay Type
Solid Phase Sandwich ELISA
Assay Range
156-10000 pg/mL
Sample Type
Cell Lysates
Reactivity
Human
Human Total FGFR4 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 FGFR4 DuoSet IC ELISA
This DuoSet IC ELISA contains the basic components required for the development of sandwich ELISAs to measure total FGF R4 in cell lysates. An immobilized capture antibody specific for FGF R4 binds both phosphorylated and unphosphorylated FGF R4. 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 FGFR4 DuoSet IC ELISA
Standard Curve
The Human Total FGF R4 DuoSet® IC ELISA is more sensitive than immunoprecipitation (IP)-Western blot analysis.
Lysates prepared from MDA-MB-453 human breast cancer cells were serially diluted and analyzed by (A) IP-Western blot and (B) this DuoSet® IC ELISA. IPs were performed using an anti-FGF R4 monoclonal antibody and goat anti-mouse agarose. Immunoblots were incubated with a biotinylated anti-FGF R4 monoclonal antibody to detect total FGF R4. Bands were visualized by chemiluminescent detection. Human FGF R4 can be detected by this DuoSet IC ELISA by using approximately 2 to 4 times less lysate than is needed for a conventional IP-Western blot.
The Human Total FGF R4 DuoSet® IC ELISA measures the relative level of FGF R4.
Lysates were prepared from MDA-MB-453 human breast cancer cells and HEK293 human embryonic kidney cells transfected with human FGF R4 (HEK293-hFGF R4). ELISA was performed using 100 μg of the MDA-MB-453 cell lysate and 5 μg of the HEK293-hFGF R4 cell lysate. IP-Western blot analyses were performed as described in Figure 1, using 400 μg of cell lysate.Kit Contents for Human Total FGFR4 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: FGFR4
Long Name
Fibroblast Growth Factor Receptor 4
Alternate Names
CD334, FGF R4
Gene Symbol
FGFR4
Additional FGFR4 Products
Product Documents for Human Total FGFR4 DuoSet IC ELISA
Product Specific Notices for Human Total FGFR4 DuoSet IC ELISA
For research use only
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