CesA4 (IRX5) | Cellulose synthase A catalytic subunit 4 [UDP-forming]
AS12 2582 | Clonality: Polyclonal | Host: Rabbit | Reactivity: Arabidopsis thaliana, Solanum lycopersicum, Solanum tuberosum
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| 500 mg of Col-0 WT Arabidopsis thaliana stem powder extracted by boiling in 2 mL of 3% SDS loading buffer + 100 mM DTT at 95C for 10 min. Extract was spun at max speed to remove debris and supernatant was taken as crude extract. 25 µL of this was loaded on a 4-15% gel run for 50 min, 150v. . Blots were blocked with 5 % milk for 1h at room temperature (RT) with agitation. Blot was incubated in the primary antibody at a dilution of 1: 500 over night at 4°C with agitation. The antibody solution was decanted and the blot was rinsed briefly twice, then washed once for 15 min and 3 times for 5 min in TBS-T at RT with agitation. Blot was incubated in secondary antibody (anti-rabbit IgG horse radish peroxidase conjugated, AS09 602 from Agrisera) diluted to 1:5000 in for 2h at RT with agitation. The blot was washed as above and developed for 5 min with high sensitivity chemiluminescent detection reagent according to the manufacturer's instructions. Exposure time was 10 seconds.
Courtesy of Dr. Manoj Kumar, University of Manchester, UK
This antibody is detecting both, recombinant and edogenous CesA4 (IRX5) protein.
CesA4 (IRX5) (EC=184.108.40.206) is a catalytic subunit of xylem-specific cellulose synthase enzyme, involved in secondary cell wall biosynthesis - cellulose synthase trerminal complexes. It interacts with CESA7 and CESA8 which is required for a functyional complex and localization in secondary cell wall deposition sites. Expressed in young plants, stems and flowers but not in leaves, roots and shoots. Amounts are increasing as stems mature.
Synonymes: Protein IRREGULAR XYLEM 5, IRX5, NWS2.
Tsuchiya et al. (2015). Distribution of XTH, expansin, and secondary-wall-related CesA in floral and fruit abscission zones during fruit development in tomato (Solanum lycopersicum). Front Plant Sci. 2015 May 15;6:323. doi: 10.3389/fpls.2015.00323.