Human Phospho-Src (Y419) DuoSet IC ELISA
R&D Systems, part of Bio-Techne | Catalog # DYC2685-2
Key Product Details
Assay Type
Solid Phase Sandwich ELISA
Assay Range
3.91-250 pg/mL
Sample Type
Cell Lysates
Reactivity
Human
Human Phospho-Src (Y419) 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 Phospho-Src (Y419) DuoSet IC ELISA
This DuoSet IC ELISA contains the basic components required for the development of sandwich ELISAs to measure phosphorylated human Src (Y419) in cell lysates. An immobilized capture antibody specific for Src binds both phosphorylated and unphosphorylated Src . 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-Src (Y419) DuoSet IC ELISA
Standard Curve
The Human Phospho-Src (Y419) DuoSet® IC ELISA specifically recognizes human Src phosphorylated at Y419 as determined by peptide competition.
A lysate prepared from MDA-MB-468 human breast cancer cells treated with 50 mM sodium pervanadate for 20 minutes was analyzed with this ELISA. The Human Phospho-Src (Y419) Detection Antibody was either untreated (no peptide) or preincubated with a phospho-peptide containing the phospho-Src Y419 phosphorylation site, a phospho-peptide containing the Ret Y905 phosphorylation site, a phospho-peptide containing the Tie-2 Y1100 phosphorylation site, or a phospho-peptide containing the PP2A Y307 phosphorylation site. Peptides were used at 40 ng/mL. Only the phospho-peptide containing the Src Y419 phosphorylation site blocked the signal, indicating that the ELISA is specific for human Src phosphorylation.
Amounts of human phosphorylated Src, as quantified by the Human Phospho-Src (Y419) DuoSet® IC ELISA, are consistent with the relative amounts of phosphorylated Src determined by qualitative Western Blot analysis.
Quantification of phosphorylated Scr (Y419) in pervabadate-treated human MDA-MB-468 cells. Lysates prepared from MDA-MB-468 human breast cancer cells treated with 50 mM sodium pervanadate for the indicated times were quantified with this DuoSet® IC ELISA. The same lysates were also immunoblotted (inset) with either Human Phospho-Src (Y419) Polyclonal Antibody (Catalog # AF2685) or Human/Mouse/Rat Src Polyclonal Antibody (Catalog # AF3389). The DuoSet® IC ELISA results are consistent with the relative amounts of phosphorylated Src family members detected by Western Blot. The immunoblot with anti-Src antibody indicated that total levels of human Src remained constant during incubation with sodium pervanadate.Kit Contents for Human Phospho-Src (Y419) 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: Src
Src and other Src family protein tyrosine kinases are proto-oncogenes that play key roles in regulating cell growth and differentiation. Src family activity is regulated by tyrosine phosphorylation at two sites with opposing effects. Phosphorylation at Y419 of human Src up-regulates kinase activity, while phosphorylation at Y530 inactivates Src.
Long Name
v-src Sarcoma [Schmidt-Ruppin A-2] Viral Oncogene Homolog
Alternate Names
ASV, c-Src, RSVgp4
Gene Symbol
SRC
Additional Src Products
Product Documents for Human Phospho-Src (Y419) DuoSet IC ELISA
Product Specific Notices for Human Phospho-Src (Y419) DuoSet IC ELISA
For research use only
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