I would like to quantify some data and use a house keeping protein which is expressed on a stable level in mouse brain after kainic acid injection (in order to induce epilepsy). We are isolating membrane proteins utilising high pH and high salt buffer. For the quantification I would use a membrane tethered protein which is expressed on a stable level even after injetion of kainic acid.
Does anyone have experience and can help me?
It is quite hard to find a good housekeeping membrane protein. I suggest you to exclude caveolin-1, as this protein is modulated after kainic acid injection at least in activated microglia. Also Na+K+ ATPase alpha subunit could be modulated after kainic acid-induced lesions. You can try with Flotillin, clathrin or Plasma membrane calcium ATPase (PMCA4). However, it is important to take into consideration that even a variety of proteins considered "housekeeper" can change after different stimuli (such as, kainic acid administration). My suggestion is to perform Coomassie Blue and/or Ponceau Red stainings, that could be used as loading control when it is not so immediate to find a good housekeeping protein in a specific experimental model.
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