Alcian Blue and Silver Staining for CPS and LPS
Posted 21 August 2012 - 04:36 AM
I'm wondering could someone help me with interpreting my SDS-PAGE gel.
I am looking for the presence of a Capsular Ploysaccharide on my bacterium, so I ran a 12% SDS-PAGE with freshly harvested cells, washed twice with PBS, and resuspended in Saline and heated at 50°C for 1hour (CPS is thermostabile) and centrifuged. The supernatant was removed and boiled with sample buffer for 5mins and loaded onto the 12% sds-gel.
I stained the gel with alcian blue and have discrete bands around 70-100kDa. There is no mAb fro the CPS of this bacteria so I can't do Western Blots.
Does anyone know how I can confirm this is a CPS or if not what else it may be?
Thanks for reading this and any advice is much appreciated!!
Posted 21 August 2012 - 12:56 PM
Posted 22 August 2012 - 08:16 AM
Thanks for your reply but unfortunately we don't have a mass spec at our ease.
Do you know from your experience if the epitopes for CPS antibodies are pretty conserved across species?