I want to do a cell adhesion assay on a 96-well Heparan Sulfate coated plate. The Heparan Sulfate should bind as naturally as possible without much changing of its conformation.
I could only find these plates: http://galenlabsupplies.com/heparin-gag-binding-plates-1/
Hi Johanna, try drying 5 micrograms of heparin sulfate solution onto the 96 well plates.
After some trying I finally used 5 micrograms as you said on High Binding ELISA plates and it worked - thank you!
Hi, Johanna,
What buffer did you prepare your 5 micrograms of heparan sulfate salt in the ELISA plates?
Thank you!
Hi Peng,
I used 200 µl of 25 µg/ml Heparan Sulfate in DPBS (1x) per well.
Best wishes
Hi, Johanna,
Thank you very much for your quick response. I wonder if you ever compare this coating protocol with other commercial heparin binding plates? And how did you determine if the coating works well?
Thanks again for your help.
Hi Peng,
as we couldn't get any Heparin binding plates in Europe (they seem to be out of stock everywhere here), we unfortunately couldn't do that. The experiments are still running, at the moment I'm comparing the heparan sulfate coating with other coatings / no coating. I'm sorry that I can't help you here!
Best wishes
Does anyone know if this same method would work to coat cell culture plates with heparan sulfate instead of ELISA plates?
As I'm also doing cell culture experiments I thought about that, too. But unfortunately you cannot analyse those with an ELISA reader because there'd be to much scattering due to special treatment for cell adhesion and sterilization procedures. If you do not want to use an ELISA reader I see no reason why it should not work just to do the coating.
Thanks Johanna. The assay won't be analysed on an ELISA reader, so fingers crossed it will work. Do you use a blocking agent, such as gelatin, after treating with Heparin sulfate?
No, but I think one cannot compare our experimental setups as I did a cell adhesion assay.
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