Am currently working on patients with various genetic cholesterol defects. We take skin biopsies and culture fibrobalsts for DNA and for in vitro testing.
To confirm the genetic finding we are trying to do a fluorescent do LDL handling/uptake study in these fibroblasts. Previous method used I125 , but we are trying not to go the radioactive route , and use di- I labelled LDL, but battling ... Any tips out there??
hi Joy, found this link, perhaps it will help
http://www.lifetechnologies.com/order/catalog/product/L3483
Hi Vincent
Thank you for your comment and suggestion - much appreciated.
My request was modified from the original, so was not a true reflection of the problem we have. We have a method from Stephan and Yurachek (J Lipid Res 34: (193) 325-329) , but although we follow the method to the letter we get very low uptake even in control fibroblasts. Perhaps the answer is not to prepare our own LDL , but to purchase the commercial one , as you suggest ... will discuss it with the team ..
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