Alpha-Tubulin Housekeeping Cellular Kit

This assay is designed to measure the endogenous expression level of alpha-tubulin in cells and tissues to normalize data obtained on the phospho- and/or total protein(s) of interest.

Due to its essential role in the cytoskeleton of cells, alpha-tubulin is ubiquitously and constitutively expressed in every tissue. Moreover, its amino acid sequence is highly conserved and is thus identical in almost all species. These characteristics make it one of the most commonly used housekeeping proteins. Its concentration is proportional to cell number and total protein concentration, and it is therefore used as an internal control for data normalization to correct for signal changes caused by experimental variability (e.g. number of cells remaining in the culture plate after treatment or lysis efficacy).The low sample volume of 4 µL and the compatibility with Cisbio’s lysis buffers #1, #2, #3 and #4 enables a multi-parametric analysis with the detection of alpha-tubulin in parallel with the phospho- and/or total protein(s) on the same lysate.

Features

  • Universal solution: works on cells and tissues of human, mouse, rat, hamster, monkey, dog and pig origin
  • Compatible with Cisbio’s lysis buffers #1, #2, #3 and #4
  • Low sample volume (only 4 µL) enabling a multi-parametric analysis on the same lysate
  • Assay range adapted to common cell densities of adherent and suspension cells
  • Useful complement to all phospho- and total protein assaysv

Applications

  • Normalization of data obtained on phospho- and/or total protein assays
  • Correction of signal changes due to experimental variability (cell number, lysis efficacy...)
  • Correct interpretation of compound effect on the protein(s) of interest
  • HTS, lead optimization, preclinical studies

Application Notes:

Assay principle

HTRF® - the homogeneous cell-based sandwich immunoassay


Cisbio Bioassay’s Alpha-Tubulin Housekeeping assay is based on a TR-FRET sandwich immunoassay format comprising two specific anti-alpha-tubulin antibodies, one labeled with Eu3+cryptate (donor) and the other with d2 (acceptor). The antibodies specifically bind to alpha-tubulin, and the proximity of donor and acceptor then leads to a fluorescent TR-FRET signal. The protocol is optimized for a low volume 96- or 384-well plate format, but can easily be further miniaturized or upscaled. Only 4 µL of sample is needed, enabling the detection of the phospho- and/or total protein of interest in parallel on the same lysate. The detection reagents may be pre-mixed and added in a single dispensing step for direct detection. No washing is needed at any step.

The assay can be run with frozen cell or tissue lysates, or with fresh cells in culture. After lysis, endogenous alpha-tubulin can be quantitatively detected using the HTRF Alpha-Tubulin Housekeeping cellular assay kit reagents and most TR-FRET multimode plate readers.

A simpler, more flexible assay protocol - adapted to your applications


Two-plate assay protocol: The assay is run under a two-plate assay protocol, where cells are plated and treated in a culture plate. For detection, 4 µL of lysate are subsequently transferred into a low volume 96- or 384-well white plate, and 12 µL of kit diluent are added before dispensing 4 µL of HTRF® reagents. This protocol enables monitoring of the cells' viability and confluence in an appropriate cell culture plate.

For more information, please visit: www.cisbio.com

Cat. No.
DescriptionAmount
64ATUBPEG
Alpha-Tubulin Housekeeping kit
500 Tests
64ATUBPEH
Alpha-Tubulin Housekeeping kit
10,000 Tests
64ATUBPEY
Alpha-Tubulin Housekeeping kit
50,000 Tests

If you would like to receive a quote for the Alpha-Tubulin Housekeeping kit, please contact our helpdesk.



Source:
Website: Cisbio Bioassays (HTRF®)

Related to:
Brands: Cisbio Bioassays (HTRF®)
Product groups: Assays