Iba 1 is used for the immunohistochemical analysis of activated microglial cells. I am doing stroke animal research and frequently encounter the staining with the Iba 1 antibody. My problem is Iba 1 stains both MCAo operated slices (suggesting the over expression of microglia in ischemia) and the sham operated slices also (though the morphology of the iba 1 positive cells from sham and MCAo are completely different.) I used both DAB substrate or immunofluoroscence for the Iba cells and got the same result. How can I eliminate the Iba 1 positive cells in sham operated (or normal) mice brain slices?
Iba-1 is constitutively expressed in the normal healthy brain. However, you should find that the level of Iba-1 is greatly enhanced following MCAO, particularly in the peri-infarct territories. In the healthy rat brain CD11b and CD68 are often less abundant that Iba-1 and you may often observe a greater relative change in these markers following injury.
Iba-1 which is a microglial marker; is constitutively expressed in the normal healthy brain. It is necessary to Iba-1 expression in control or sham brain. Since it is not inducible in nature but it get activated in pathological condition. How ever if you want to compare only in MCAO model. You can cut the tissue in two parts like right hemisphere and left hemisphere. I mean to say non affected area-contra lateral and affected area ipsi lateral. In that way you can also present your data for Iba-1 expression. Good luck.
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