Anti-RGL2 | RGA-like protein 2 (DELLA protein)

Brassica napus, Populus sp., Triticum sp.

Samples:
1- 50 µg/well of Arabidopsis thaliana, ecotype Lens-A whole 6-week-old plant extract.
2- 50 µg/well of Pentuple mutant of all five DELLA genes (RGA, GAI, RGL1, RGL2 and RGL3) generated by genetic cross; (CS16298) 6-week-old plant extract.
3- 50 µg/well of Arabidopsis thaliana, ecotype Lens-A whole 3-week-old seedling extract. 4- 50µg/well of Pentuple mutant of all five DELLA genes (RGA, GAI, RGL1, RGL2 and RGL3) generated by genetic cross; (CS16298) 3-week-old seedling extract.
Markers: PAGEmark Dual color Protein Marker from G-Biosciences

50 µg/well of total protein extracted freshly from 6 to 3-week-old Arabidopsis thaliana seedlings with extraction buffer (10 mM Tris-HCl pH 7.5, 150 mM NaCl, 10 mM MgCl₂, 1 mM EDTA and 1x protease inhibitor cocktail) and denatured with with 1X SDS-Dye at 95°C for 5 min. Samples were separated in the cold on 8 % SDS-PAGE and blotted for 1.5 h to nitrocellulose (pore size of .2 µm), using wet transfer in the cold. Blot was blocked with 5 % milk for 1h/RT with agitation. Blot was incubated in the primary antibody at a dilution of 1: 500 for ON/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 PBS-T at RT with agitation. Blot was incubated in matching secondary antibody (anti-rabbit IgG horse radish peroxidase conjugated, AS09 602 Agrisera) diluted to 1: 10 000 in 1X PBS-T for 1 h/RT with agitation. The blot was washed as above and developed with a following. The blot was washed as above and developed for 3 min with chemiluminescence detection reagent before the image was captured.

Courtesy of phd student Priyanshi Rana, Department of Biochemistry Indian Institute of Science, India

As this protein is expressed in tissue in very low levels, please load at least 50 µg/well.
Recommended extraction and denaturation protocol: 

DELLA protein RGL2 is a probable transcriptional regulator that acts as a repressor of the gibberellin (GA) signaling pathway. Involved in the biosynthesis of abscisic acid (ABA), delays flowering and leaf production, reulates floral development, petal, stamen and anther development. Predominantly expressed in germinating seeds, flowers and siliques.  Synonymes: GRAS family protein 15, AtGRAS-15, Scarecrow-like protein 19, AtSCL19.