I am trying to do DDR1 receptor endocytosis test. I use collageI coated fask to activate DDR1 and want to know if DDR1 will be phophorylated and be endocytosed in to the cell. how can I exclude the phophorylated DDR1 in adhesions when I try to extract proteins for western-blot? Thank you.
Dear Zheng,
Did you try to extract the "cytosolic" fraction by adding detergent-free ice-old hypotonic buffers (plus inhibitor cocktails). after taking this extract you can proceed to scratch the dish with regular RIPA buffer so you can compare these 2 extracts in control and treated conditions. If your treatment cause the receptor's endocytosis, then you should see the increased receptor levels in the hypotonic extract of the treated condition.
Hope it may help,
Dear Sebastián Dupraz ，
Thank you so much, I have not tried that. I will get the information and try. Thank you very much.
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