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Our Abpromise guarantee covers the use of ab86408 in the following tested applications.
|IHC-FoFr||Use at an assay dependent concentration.|
|WB||Use a concentration of 1 µg/ml. Predicted molecular weight: 36 kDa. Good results were obtained when blocked with 5% non-fat dry milk in 0.05% PBS-T.|
Male mouse (P41) was anesthetized with isoflurane until they became non-responsive to tail pinch. Animal's thoracic cavity opened and it was transcardially perfused with 10 mL of ice cold PBS, then 10 mL of Ice cold 4 % PFA solution. Brain was removed and fixed overnight in 4% PFA. Next morning brain was rinsed 3X in PBS. Brain was immersed in agar and sliced at a thickness of 50uM. Brain slices were incubated overnight in a solution of PBST (0.2%) + 10% Normal GOAT serum + a monoclonal mouse anti-Parvalbumin antibody (1:2000)+ ab86408 (1ug/mL) and gently shaken overnight at 4°C. Samples were washed 6x in PBS (10 min each at RT) and exposed to secondary antibodies at 4°C: PBST (0.3%) + 10% normal goat serum + anti-mouse Alexa 488 (1:1000) + Anti-rabbit Alexa 555 (1:1000). Samples were washed 6x in PBST mounted. Images acquired with a confocal microscope at 63X.