Anti-Neu5Gc Antibody Kit Protocol - Western blot
Prepare two identical SDS-PAGE gels. 
Load and run samples in loading buffer. 
Recommended loading of gel: 
Lane 1: Molecular Weight Marker.
Lanes 2, 8, 10: Blank.
Lanes 3-6: Samples.
Lane 7: Positive Control.
Lane 9: Negative Control. 
Confirm the presence of proteins on gel by Coomassie Staining Blot proteins to membrane of choice. 
Confirm protein transfer by Ponceau or other method of choice. 
Block each membrane in 20 ml TBS-T with 200 μl diluted Neu5Gc Assay Blocking Solution, gently rocking at 4°C.  
Incubate first blot in Primary Antibody, and one blot in Control Antibody for 2 hours at 25°C or overnight at 4°C with gentle rocking. 
Wash blot with 50 ml TBS-T for 5 min at 25°C, gently rocking 5 times.
Incubate blots with optimal dilution of enzyme-conjugated secondary antibody of choice at 25°C for 1 hour. 
Wash blot with 50 ml TBS-T for 5 min at 25°C , gently rocking 5 times.
Develop each blot with the appropriate substrate
