This HTRF kit is designed for the rapid detection of total alpha tubulin 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).
Feature | Specification |
---|---|
Application | Protein Quantification |
Sample Volume | 16 µL |
This HTRF kit is designed for the rapid detection of total alpha tubulin 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).
Alpha-Tubulin is a member of the tubulin protein superfamily, which polymerizes with the beta tubulin to form microtubules, a major component of the eukaryotic cytoskeleton. Microtubules contribute to several cellular processes, such as structural support, intracellular transport, and DNA segregation. The Alpha-Tubulin kit is designed for the quantitative detection of total Alpha-Tubulin in cell lysates. This assay can be used as a normalization readout.
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 |
Oncology & Inflammation
|
Unit Size |
500 Assay Points
|
Alpha-Tubulin is measured using a sandwich immunoassay involving two specific anti-alpha-Tubulin antibodies, respectively labelled with Europium Cryptate (donor) and d2 (acceptor). The intensity of the signal is proportional to the concentration of the total alpha-Tubulin present in the sample.
The protocol for the Alpha-Tubulin assay is described here. Cells are plated, stimulated, and lysed in the same 96-well culture plate. Lysates are then transferred into the assay plate for the detection of alpha-Tubulin by HTRF reagents. 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.
12,500 NIH/3T3 cells were plated in 96-well plate and treated after incubation overnight for 16h with compounds at 37°C, CO2. Lysis buffer was added after supernatant removal. 30 minutes under shaking conditions permit the lysate generation and then we transferred the lysatesd to 384-well plate for detection. Trichostatin A (TSA), a potent inhibitor of histone deacetylase, and Tubacin, a selective inhibitor of HDAC6, increase the level of acetylated α –Tubulin on Lysine 40 on NIH/3T3 cells while the expression level of α –Tubulin in presence of both inhibitors remains constant.
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This guide provides you an overview of HTRF applications in several therapeutic areas.
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