Western blot detection other than antibody? - (Mar/02/2005 )
Anyone know how to detect and quantify protein in western blot using method other than HRP-antibody? My lab is in tight budget and we are looking for cheaper alternative. Thanx heaps!
you can use alkaline-phosphatase (AP) coupled antibodies, but I'm not sure if this is cheaper than HRP, taken together the cost for the Ab and the substrate....
Je suis pas sûre que ça convienne mais c'est vraiment le moins coûteux: tu peux faire 1 coloration au rouge Ponceau ou ou bleu de Coomassie et regarder si tu as 1 bande à la taille que tu attends! Ensuite, si ce prétest est positif tu utilise les anticorps (moi aussi je prends des coumplés à la Phosphatase alcaline) pour confirmer la nature de ta protéine. Tu ne les utilises pas pour rien au moins. Bonne chance!
Errr, my french is a little bit rusty (i.e. plus mal), and there wasn't much french to begin with, so maybe people here can understand this way better than me, so corrections welcome!
I think margo33 says, you can stain your membrane with coommassie or ponceau and then use comparing methods to determine your protein-concentrations, besides she would use AP-antibodies, too, and wishes you luck!
(I hope this is halfway correct )
I wanted to say: you can stain your membrane with coommassie or ponceau and look if you've got a band at the size you 're waiting for. Then you can use AP-antibodies to confirm it's the right protein. So you don't waste your antibody for nothing!
Sorry for the answer in french!
I'd say: "swing and a miss", but a close one, sorry!