Pro q emerald glycoprotein gel and blot stain kit

LdMVK protein was electrophoresed (SDS-PAGE) and staining of glycoprotein was carried out as stated in Pro-Q Emerald glycoprotein gel and blot stain kit manufacturer’s protocol (Molecular Probes, Eugene, Oreg). Protein in gel was fixed in a solution of 50% methanol and 5% acetic acid and washed in 3% acetic acid solution. Gel was incubated in oxidizing solution provided in the kit for 30 min and washed two times. Incubation in Pro-Q Emerald 300 staining solution was done for 2 h and stained gel was then visualized by illuminating in UV light in BioRad transilluminator. CandyCane™ glycoprotein molecular weight standards (Molecular Probes, Eugene, OR, USA) containing a mixture of alternate glycosylated and non-glycosylated proteins was separated by SDS-PAGE and stained by Pro-Q Emerald glycoprotein gel and blot stain kit, and it served as positive and negative control for the experiment. The staining was performed at least three times and protein from different batches were used each time.