Brain tissue homogenates and IL-1 - (Nov/24/2008 )
Hi, Im very new in ELISA so I have a lot of questions. Anyway, Im doing sandwich ELISA with brain tissue homogenates for IL-1, the minimum detectability for this kit (e-Bioscience) is 39 pg, and I didnt get a detectable level of IL-1 in rat serum, but when i use it in brain homogenates, i get actually very high values for ex. 2400 pg/mg total proteins (i put 125 microgr in 50 microl/per well). Probably Im doing something wrong, i dont know what kind of control can i use, if the company claims there is no cross-reactivity with other molecules how i can know if these are false results?
Thank you in advance
Natasa
-natasav-
QUOTE (natasav @ Nov 24 2008, 06:07 AM)
Hi, Im very new in ELISA so I have a lot of questions. Anyway, Im doing sandwich ELISA with brain tissue homogenates for IL-1, the minimum detectability for this kit (e-Bioscience) is 39 pg, and I didnt get a detectable level of IL-1 in rat serum, but when i use it in brain homogenates, i get actually very high values for ex. 2400 pg/mg total proteins (i put 125 microgr in 50 microl/per well). Probably Im doing something wrong, i dont know what kind of control can i use, if the company claims there is no cross-reactivity with other molecules how i can know if these are false results?
Thank you in advance
Natasa
Thank you in advance
Natasa
Just to add, tissue was homogenized in PBS+0.1% Triton X-100+p. inhibitors and cfg. on 15.000g/20 min, supernatant (125 microg in 50 microl) was undiluted used for ELISA
-natasav-