HTRF acetyl K40 α-Tubulin Detection Kit, 500 Assay Points

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HTRF acetyl K40 α-Tubulin Detection Kit, 500 Assay Points

A-tubulin Acetyl-k40 Kit - 100,000 Tests

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

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A-tubulin Acetyl-k40 Kit - 100,000 Tests

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Product Variants

Unit Size: 500 Assay Points

Part #:

63ADK072PEG

List Price

USD 1,253.00

Unit Size: 10,000 Assay Points

Part #:

63ADK072PEH

List Price

USD 8,183.00

Product information

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.

Specifications

Other Specifications

Application	Protein Quantification
Brand	HTRF
Detection Modality	HTRF
Molecular Modification	Acetylation
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

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How it works

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.

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.

Assay validation

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.