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Alpha-Tubulin acetyl-K40 kit HTRF®

The Alpha-Tubulin acetyl-K40 kit is designed for the rapid detection of K40 tubulin acetylation in cell lysates.
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  • No-wash No-wash
  • Low sample consumption Low sample consumption
  • High sensitivity High sensitivity
The Alpha-Tubulin acetyl-K40 kit is designed for the rapid detection of K40 tubulin acetylation in cell lysates.
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Overview

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. Alpha-Tubulin K40 acetylation has been shown to affect tubulin self-assembly and protect microtubules against stress-induced material fatigue. The Alpha-Tubulin acetyl-K40 kit is designed for the quantitative detection of K40 acetylated Alpha-Tubulin in cell lysates.

Benefits

  • HDAC6 MODULATION STUDY

Assay principle

Alpha-Tubulin acetyl-K40 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 K40 acetylated alpha-Tubulin present in the sample.
Alpha tubullin kit principle

Assay Protocol

The protocol for the alpha-Tubulin acetyl-K40 assay is described here. Cells are plated, stimulated, and lysed in the same 96-well culture plate. Lysates are then transferred to 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.
Alpha tubulin kit protocol

TSA and Tubacin effect on NIH 3T3 cells

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.
TSA and Tubacin effect on NIH 3T3 cells

HTRF Product Catalog

All your HTRF assays in one document! - Catalog

A guide to Homogeneous Time Resolved Fluorescence

General principles of HTRF - Guides

How HTRF compares to Western Blot and ELISA

Get the brochure about technology comparison. - Brochures

Plate Reader Requirement

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