Human IL-17RB DuoSet ELISA

Catalog #: DY1207 Datasheet / COA / SDS

Catalog #	Availability	Size / Price	Qty
DY1207

Ancillary Products Available

DY008B: DuoSet ELISA Ancillary Reagent Kit 2

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All IL-17RB ELISAs

Product Details

Procedure

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

Human IL-17RB DuoSet ELISA Summary

Assay Type

Solid Phase Sandwich ELISA

Format

96-well strip plate

Sample Volume Required

100 µL

Assay Range

156.0 - 10,000 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 IL-17 RB. 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 IL-17 RB 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: IL-17RB

IL-17B receptor (IL-17B R), also known as IL-17Rh1, IL-17ER and EVI27, is a 502 amino acid (aa) type I membrane protein with a 17 aa signal peptide, a 275 aa extracellular domain, a 21 aa transmembrane domain and a 189 aa cytoplasmic tail. By alternative splicing, a secreted variant of IL-17B R has also been identified.

Long Name:

Interleukin 17B Receptor

Entrez Gene IDs:

55540 (Human); 50905 (Mouse)

Alternate Names:

CRL4IL-17 receptor homolog 1; cytokine receptor CRL4; Cytokine receptor-like 4; Evi27; EVI27IL-17B receptor; IL-17 RB; IL-17 receptor B; IL-17 RH1; IL17BRinterleukin 17B receptor; IL-17ER; IL17RB; IL-17RB; IL17Rh1; IL-17Rh1; IL17RH1MGC5245; interleukin 17 receptor B; interleukin 17 receptor homolog 1; interleukin-17 receptor B; Interleukin-17B 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.