Alpha-Tubulin Housekeeping standard

This standard is a spare reagent for the Alpha-Tubulin Housekeeping kit. It may be used for absolute quantification of cellular alpha-tubulin extracted with Cisbio’s lysis buffer.

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This standard is a spare reagent for the Alpha-Tubulin Housekeeping kit. It may be used for absolute quantification of cellular alpha-tubulin extracted with Cisbio’s lysis buffer.

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Overview

This standard provides a way to quantify the alpha tubulin contained in your sample

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Correlation with the BCA protein assay

Lysates of human and mouse cell lines were plated at different cell densities in 96-well culture plates for 5 hours at 37°C, 5% CO2. Cells were then lysed with 50 µL of supplemented lysis buffer #4 for 30 minutes at RT under gentle shaking, and 4 µL of cell lysate were transferred into a low volume white microplate. For HTRF detection, 12 µL of kit diluent were added followed by the dispensing of 4 µL of premixed detection antibodies. The HTRF signal was recorded after an overnight incubation at RT. In parallel, the concentration of proteins was determined in the same lysates using the BCA protein assay (QuantiPro™ BCA Assay Kit, Sigma-Aldrich) according to the manufacturer’s instructions. For both methods (HTRF or BCA), lysates were analyzed either neat or prediluted in the appropriate buffer in order to work within the linear range of each kit. The cell densities used for the correlations are mentioned on each graph next to each point.