I need to do western blot of RBC globin but i can't find how to extract globin from the cells. I will use anti-AGE antibody. Did somebody do something like that? Thank everybody for answers!
Hi Taras, are you talking about extracting hemoglobin from RBCs of diabetic patients?
If so, then there are specific antibodies for the glycated form of hemoglobin (HbA1c). I don't think anti AGE antibodies will work well on glycated hemoglobin, because it usually does not develop advanced glycation end products in vivo.
But if you want to isolate glycated hemoglobin from RBC, then I recommend using immobilized boronic acid.
http://www.piercenet.com/product/boronic-acid-resin
Hi Steingrimur! thank you for your answer. But:
1. Yes, I work with RBCs of diabetic, but not patients. with rats.
2. I don't understand fully what do you mean when you say "because it usually does not develop advanced glycation end products in vivo".
3. I want to extract not hemoglobin but globin without hem-group.
Hi Taras, point taken.
I was thinking that you were analyzing hemoglobin from diabetic patients.
What I meant with Hb that "it usually does not develop advanced glycation end products in vivo." is that the RBCs have a high turnover in the circulation (20-30 days) that precludes them from developing more advanced Amadori and Maillard reaction products that can be classified as AGE.
You usually don't find vascular proteins with AGE products because vascular proteins are turned over rapidly by the liver.
Extravascular proteins, like collagens and elastin, however, have a long turnover rate (~years), are more likely to develop AGE products.
Question. Why do you want to want to remove the heme group from hemoglobin? The heme is not involved in the glycation process as far as I know.
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