Human EDA2R/TNFRSF27/XEDAR DuoSet ELISA

Catalog #: DY1093 Datasheet / COA / SDS

Catalog #	Availability	Size / Price	Qty
DY1093

Ancillary Products Available

DY008B: DuoSet ELISA Ancillary Reagent Kit 2

Human EDA2R / TNFRSF27 / XEDAR ELISA Standard Curve

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Procedure

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

Human EDA2R/TNFRSF27/XEDAR DuoSet ELISA Summary

Assay Type

Solid Phase Sandwich ELISA

Format

96-well strip plate

Sample Volume Required

100 µL

Assay Range

15.6 - 1,000 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 human EDA2R / TNFRSF27 / XEDAR. 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 ELISA.

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

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 equivalent

Reagent Diluent: 1% BSA in PBS, pH 7.2-7.4, 0.2 m filtered (R&D Systems Catalog # DY995). Quality of BSA is critical (see Technical Hints).

Blocking Buffer: 1% BSA in PBS, pH 7.2-7.4, 0.2 m filtered (R&D Systems Catalog # DY995). Quality of BSA is critical (see Technical Hints).

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

Scientific Data

N/A

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Human EDA2R / TNFRSF27 / XEDAR 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: EDA2R/TNFRSF27/XEDAR

EDA2R, also known as TNFRSF27 or XEDAR (X-linked ectodysplasin receptor), is an X-linked member of the TNF receptor superfamily (TNFRSF). Within the TNFRSF, EDA2R shares the highest homologies with EDAR and TNFRSF19/TROY. EDA-A2 is the EDA2R ligand. EDA2R expression is principally found in embryonic hair follicles. EDA2R, EDAR, EDA-A1, and EDA-A2 have been associated with hypohidrotic ectodermal dysplasia (HED).

Long Name:

Ectodysplasin A2 Receptor

Entrez Gene IDs:

60401 (Human); 245527 (Mouse); 710244 (Rhesus Macaque)

Alternate Names:

ectodysplasin A2 receptor; EDA2R; EDA-A2 receptor; EDAA2R; EDA-A2R; EDAA2Rtumor necrosis factor receptor superfamily member XEDAR; EDAR2; TNFRSF27; XEDAR; XEDARTNFRSF27tumor necrosis factor receptor superfamily member 27; X-linked ectodysplasin-A2 receptor

Assay Procedure

GENERAL ELISA PROTOCOL

Plate Preparation

Dilute the Capture Antibody (to the working concentration stated in the product datasheet ) 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 each well of the microplate as recommended in the product datasheet. Incubate at room temperature for a minimum of 1 hour.

Note: The recommended Reagent Diluent typically contains 1% BSA. Some DuoSet Development Kits require alternative blocking agents, or for plates to be blocked overnight with a higher percentage of BSA, please see the product datasheet for details.
Repeat the aspiration/wash as in step 2. The plates are now ready for sample addition.

PRECAUTION
The Stop Solution suggested for use with this kit is an acid solution. Wear eye, hand, face and clothing protection when using this material.

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 (as recommended in the product datasheet), 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.