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

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  • All inclusive kit All inclusive kit
  • Highly specific Highly specific
  • Faster and more convenient than ELISA Faster and more convenient than ELISA

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

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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. Cisbio's Utrophin kit is designed for its rapid detection in cell lysates.​​​​​​​

Benefits

  • Cost-effective alternative to ELISA
  • Measures Mouse and Human Utrophin
  • Validated on HeLa and C2C12 cells

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.
Principle of the HTRF Utrophin assay

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.

2-plate protocol of the HTRF Utrophin assay

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.

Image of the HeLa cell density using the 2-plate assay protocol of the HTRF Utrophin assay

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.

Image of the C2C12 cell density using the 2-plate assay protocol of the HTRF Utrophin assay
Cisbio Product Catalog 2019

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A guide to Homogeneous Time Resolved Fluorescence

General principles of HTRF - Guides

How HTRF compares to Western Blot and ELISA

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Product Insert Utrophin Kit / 63ADK107PEG-63ADK107PEH

63ADK107PEG-63ADK107PEH - Product Insert

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