Human Phospho-Src (Y419) DuoSet IC ELISA

Name: Human Phospho-Src (Y419) DuoSet IC ELISA
Brand: R&D Systems
SKU: DYC2685-2
Price: 495 USD
Availability: InStock

R&D Systems | Catalog # DYC2685-2

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Product Summary
Product Specifications
Scientific Data
Kit Contents
Other Reagents Required
Preparation & Storage
Background
Product Documents
Specific Notices
Research Areas

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

View all Src ELISA Kits »

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

Please see the product datasheet

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.