Cp2 | Cysteine protease
AS16 3110 | Clonality: Polyclonal | Host: Rabbit | Reactivity: Carica papaya
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Total proteins from Carica papaya cultivar Eksotika and MARDI Purple leaves were extracted using pre-cooled extraction buffer containing 50 mM phosphate buffer pH7 and protease inhibitor cocktail. 60 µg of total protein was blotted 1 h to PVDF using semi-dry transfer cell (BioRad, USA). Blots were blocked with 3% milk in PBST for overnight at 4°C with agitation. Blot was incubated in the primary antibody at a dilution of 1: 1 000 in 1% BSA/PBST for 3h at 4°C with agitation. The antibody solution was decanted and the blot was washed 3 times for 5 min each in PBS at RT with agitation. Blot was incubated in secondary antibody (anti-rabbit IgG horse radish peroxidase conjugated, AS09 602, Agrisera) diluted to 1:20 000 in 1% BSA/PBST for 2 h at RT with agitation. The blot was washed as above and stained with Amplified Opti-4CN Substrate kit (BioRad, USA).
Apparent MW of Cp2 is 40 kDa.
Courtesy of Suhaina Supian, Malaysian Agricultural Research and Development Institute, Malaysia
Cysteine protease Cp2 shows proteolytic activity and is involved in programmed cell death in plant during pathogen infection.
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