Western blot detection other than antibody? - (Mar/02/2005 )
Hi guys,
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! 
Sissle
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....
mike
Je suis pas s�re que �a convienne mais c'est vraiment le moins co�teux: tu peux faire 1 coloration au rouge Ponceau ou ou bleu de Coomassie et regarder si tu as 1 bande � la taille que tu attends! Ensuite, si ce pr�test est positif tu utilise les anticorps (moi aussi je prends des coumpl�s � la Phosphatase alcaline) pour confirmer la nature de ta prot�ine. 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
)mike
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!
@margo:
I'd say: "swing and a miss", but a close one, sorry! 
mike