This HTRF kit is designed for the rapid detection of total alpha tubulin in cell lysates.
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.
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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