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HTRF Utrophin Detection Kit, 500 Assay Points

The HTRF Utrophin kit is designed for the rapid detection of Utrophin in cell lysates.

For research use only. Not for use in diagnostic procedures. All products to be used in accordance with applicable laws and regulations including without limitation, consumption and disposal requirements under European REACH regulations (EC 1907/2006).

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Feature Specification
Application Protein Quantification
Sample Volume 16 µL

The HTRF Utrophin kit is designed for the rapid detection of Utrophin in cell lysates.

For research use only. Not for use in diagnostic procedures. All products to be used in accordance with applicable laws and regulations including without limitation, consumption and disposal requirements under European REACH regulations (EC 1907/2006).

Click to copy promo code to clipboard.
SAVE 15% NOW on online orders. Use promo code below.
YEAREND15
Offer valid until 12/15. Terms and conditions apply.
Product Variants
Unit Size: 500 Assay Points
Part #:
63ADK107PEG
List Price
USD 1,161.00
Unit Size: 10,000 Assay Points
Part #:
63ADK107PEH
List Price
USD 13,640.00

Overview

Utrophin is an autosomally-encoded 376-kDa cytoskeletal protein that is similar in structure and function to dystrophin. It is a ubiquitously expressed protein that plays a role in anchoring the cytoskeleton to the plasma membrane. Utrophin is highly expressed in developing muscle, and is enriched at the neuromuscular junction of mature muscle. Duchenne's muscular dystrophy patients lack dystrophin, and utrophin is consequently up-regulated and redistributed to locations normally occupied by dystrophin. HTRF Utrophin kit is designed for its rapid detection in cell lysates.​​​​​​​

Specifications

Application
Protein Quantification
Brand
HTRF
Detection Modality
HTRF
Product Group
Kit
Sample Volume
16 µL
Shipping Conditions
Shipped in Dry Ice
Target Class
Biomarkers
Technology
TR-FRET
Therapeutic Area
Rare Diseases
Unit Size
500 Assay Points

Video gallery

How it works

Utrophin assay principle

Utrophin is measured using a sandwich immunoassay involving two specific anti-utrophin antibodies, respectively labelled with Europium Cryptate (donor) and d2 (acceptor). The intensity of the signal is proportional to the concentration of utrophin present in the sample.

1biomarkers-how-it-works-assay-principle-utrophin-63adk107peg-63adk107peh.svg

 

Utrophin assay protocol

The Utrophin assay features a two-plate assay protocol, where cells are first plated, stimulated, and lysed in the same culture plate. Lysates are then transferred to the assay plate for the detection of utrophin. This protocol enables the cells' viability and confluence to be monitored. The antibodies labelled with HTRF fluorophores may be pre-mixed and added in a single dispensing step to further streamline the assay procedure. The assay detection can be run in 96- to 384-well plates by simply resizing each addition volume proportionally.

2biomarkers-how-it-works-assay-protocol-alpha-tubulin-63adk072peg-63adk072peh.svg

 

Assay validation

Endogenous level of utrophin in HeLa cells

Human HeLa cells were plated under 100 µl in 96-well plates at different cell densities (100K, 50K, 25K, 12.5K, 6.25K, and 3.1K cells/well) in complete culture medium, and incubated at 37°C, 5% CO2. The day after, medium was removed and cells were then lysed with 50 µL of lysis buffer #3 for 30 minutes at room temperature under gentle shaking. Next, 16 µL of lysate were transferred into a low volume white microplate before the addition of 4 µL of the premixed HTRF Utrophin detection reagents. The HTRF signal was recorded after ovenight incubation.

2assay-validation-utrophin-1.svg

 

Endogenous level of utrophin in C2C12 cells

Mouse C2C12 cells were plated under 100 µl in a 96-well plates at different cell densities (200K, 100K, 50K, 25K, 12.5K, and 6.25K cells/well) in complete culture medium and incubated at 37°C, 5% CO2. The day after, medium was removed and cells were then lysed with 50 µL of lysis buffer #3 for 30 minutes at room temperature under gentle shaking Next, 16 µL of lysate were transferred into a low volume white microplate before the addition of 4 µL of the premixed HTRF Utrophin detection reagents. The HTRF signal was recorded after overnight incubation.

3assay-validation-utrophin-2.svg

 

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HTRF solutions, guide to major applications

This guide provides you an overview of HTRF applications in several therapeutic areas.

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