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Fig. 2 | Applied Biological Chemistry

Fig. 2

From: Production, characterization, and cross-reactivity of a polyclonal antibody against Arabidopsis TARGET OF RAPAMYCIN

Fig. 2

Preparation and concentration of recombinant His-TOR1−200 and His-TOR1113−1304. a Workflow depicting the procedure for producing cell lysates expressing His-TOR1−200 and His-TOR1113−1304 to concentrate the Ppt fractions. b Preparation of antigens for concentrated Ppt fractions. Ppt fractions containing His-TOR1−200 and His-TOR1113−1304 isolated from cell lysates were subjected to several rounds of sonication and divided into Ppt and Sup fractions as described in a. The proteins were separated in 10% SDS-PAGE. c Concentration estimates of His-TOR1−200 and His-TOR1113−1304. Ppt fractions as prepared in b were rerun in 10% SDS-PAGE, and the bands corresponding His-TOR1−200 and His-TOR1113−1304 were excised from gels. The proteins were eluted from gel strips using the electro-eluter and run in 10% SDS-PAGE. Eluted proteins were collected by three different fractions (top, middle, and bottom) in a collection tube. BSA was used as a marker protein to estimate the protein concentrations. Molecular size markers (kDa) are represented on the left

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