Mouse EGF DuoSet ELISA

Catalog #: DY2028 Datasheet / COA / SDS

Catalog #	Availability	Size / Price	Qty
DY2028

Ancillary Products Available

DY008B: DuoSet ELISA Ancillary Reagent Kit 2

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All EGF ELISAs

Product Details

Procedure

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Summary Product Features Kit Content Other Reagents Required Data Examples Product Datasheets Preparation and Storage Background

Mouse EGF DuoSet ELISA Summary

Assay Type

Solid Phase Sandwich ELISA

Format

96-well strip plate

Sample Volume Required

100 µL

Assay Range

3.9 - 250 pg/mL

Sufficient Materials

For fifteen 96-well plates*

Specificity

Please see the product datasheet

* Provided that the recommended microplates, buffers, diluents, substrates and solutions are used, and the assay is run as summarized in the Assay Procedure provided.

This DuoSet ELISA Development kit contains the basic components required for the development of sandwich ELISAs to measure natural and recombinant mouse EGF. The suggested diluent is suitable for the analysis of most cell culture supernate samples. Diluents for complex matrices, such as serum and plasma, should be evaluated prior to use in this DuoSet.

Product 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
Economical alternative to complete kits

Kit Content

Capture Antibody
Detection Antibody
Recombinant Standard
Streptavidin conjugated to horseradish-peroxidase (Streptavidin-HRP)

Other Reagents Required

DuoSet Ancillary Reagent Kit 2 (5 plates): (Catalog # DY008) containing 96 well microplates, plate sealers, substrate solution, stop solution, plate coating buffer (PBS), wash buffer, and Reagent Diluent Concentrate 2.

The components listed above may be purchased separately:

PBS: (Catalog # DY006), or 137 mM NaCl, 2.7 mM KCl, 8.1 mM Na₂HPO₄, 1.5 mM KH₂PO₄, pH 7.2 - 7.4, 0.2 µm filtered

Wash Buffer: (Catalog # WA126), or 0.05% Tween^® 20 in PBS, pH 7.2-7.4

Reagent Diluent: (Catalog # DY995), or 1% BSA in PBS, pH 7.2-7.4, 0.2 µm filtered

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)

Plate Sealers: ELISA Plate Sealers (Catalog # DY992)

Scientific Data

N/A

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Mouse EGF ELISA Standard Curve

Product Datasheets

Product Datasheet

COA

SDS

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: EGF

EGF (Epidermal Growth Factor) is expressed as a transmembrane protein that is proteolytically cleaved to generate soluble forms. It signals through EGF R/ErbB1 which heterodimerizes with ErbB2, ErbB3, or ErbB4 and can also associate with PDGF R beta and HGF R/c-MET. During development, EGF regulates thymocyte differentiation, neuroglia production, and adipocyte maturation. In the adult, EGF plays a role in mammary gland lactogenesis, fibroblast mitosis, dissociation of the extracellular matrix, and cell migration.

Long Name:

Epidermal Growth Factor

Entrez Gene IDs:

1950 (Human); 13645 (Mouse); 25313 (Rat)

Alternate Names:

beta-urogastrone; EGF; epidermal growth factor (beta-urogastrone); epidermal growth factor; hEGF; HOMG4; pro-epidermal growth factor; URG; Urogastrone

Assay Procedure

GENERAL ELISA PROTOCOL

Plate Preparation

Dilute the Capture Antibody to the working concentration in PBS without carrier protein. Immediately coat a 96-well microplate with 100 μL per well of the diluted Capture Antibody. Seal the plate and incubate overnight at room temperature.
Aspirate each well and wash with Wash Buffer, repeating the process two times for a total of three washes. Wash by filling each well with Wash Buffer (400 μL) using a squirt bottle, manifold dispenser, or autowasher. Complete removal of liquid at each step is essential for good performance. After the last wash, remove any remaining Wash Buffer by aspirating or by inverting the plate and blotting it against clean paper towels.
Block plates by adding 300 μL Reagent Diluent to each well. Incubate at room temperature for a minimum of 1 hour.
Repeat the aspiration/wash as in step 2. The plates are now ready for sample addition.

Assay Procedure

Add 100 μL of sample or standards in Reagent Diluent, or an appropriate diluent, per well. Cover with an adhesive strip and incubate 2 hours at room temperature.
Repeat the aspiration/wash as in step 2 of Plate Preparation.
Add 100 μL of the Detection Antibody, diluted in Reagent Diluent, to each well. Cover with a new adhesive strip and incubate 2 hours at room temperature.
Repeat the aspiration/wash as in step 2 of Plate Preparation.
Add 100 μL of the working dilution of Streptavidin-HRP to each well. Cover the plate and incubate for 20 minutes at room temperature. Avoid placing the plate in direct light.
Repeat the aspiration/wash as in step 2.
Add 100 μL of Substrate Solution to each well. Incubate for 20 minutes at room temperature. Avoid placing the plate in direct light.
Add 50 μL of Stop Solution to each well. Gently tap the plate to ensure thorough mixing.
Determine the optical density of each well immediately, using a microplate reader set to 450 nm. If wavelength correction is available, set to 540 nm or 570 nm. If wavelength correction is not available, subtract readings at 540 nm or 570 nm from the readings at 450 nm. This subtraction will correct for optical imperfections in the plate. Readings made directly at 450 nm without correction may be higher and less accurate.