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Fig. 4 | Biological Procedures Online

Fig. 4

From: Fluorescence ImmunoPrecipitation (FLIP): a Novel Assay for High-Throughput IP

Fig. 4

Evaluation of the amount of cells necessary for reliable FLIP. HeLa cells were plated, transfected and lysed in wells of different sizes. Lysates from each well were used to perform FLIP and IP/IB. a FLIP signal (total mean fluorescence measured by ImageJ) using a FLAG Ab subtracted from the FLIP signal using control IgG antibodies (background) is reported. Two FLIP measurements from 2 different aliquots of beads were used to calculate the range of variability of the measurement (var.). b The immunoprecipitated beads not used for FLIP were analyzed by immunoblotting. LY = input lysate; IgG = control IP using IgG antibodies; FLAG = IP of the tagged protein using anti FLAG antibodies. Tubulin was used as loading control

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