FW: Sandwich ELISA with brain tissue homogenates - Question asked by Natasa (Nov/24/2008 )
Hi,
Natasa had PMed me about her question about sandwish ELISA with brain tissue homogenates.
Now I put her PM to this forum.
Hope someone who have more experience with tissue homogenates in ELISA can help her.
"Dear Minnie, excuse me for writing to your private adress, but I think maybe be you can help me with ELISA problem. 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. Probably Im doing something wrong, i dont know what kind of control can i use, if the company claims there is no croos-reactivity with other molecules how i can know if these are false results?
Thank you in advance
Natasa
I will send you my raw results"
http://www.protocol-online.org/forums/inde...msg&id=5593
can you get IL-1 knockout rat?
Natasa had PMed me about her question about sandwish ELISA with brain tissue homogenates.
Now I put her PM to this forum.
Hope someone who have more experience with tissue homogenates in ELISA can help her.
"Dear Minnie, excuse me for writing to your private adress, but I think maybe be you can help me with ELISA problem. 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. Probably Im doing something wrong, i dont know what kind of control can i use, if the company claims there is no croos-reactivity with other molecules how i can know if these are false results?
Thank you in advance
Natasa
I will send you my raw results"
http://www.protocol-online.org/forums/inde...msg&id=5593
Natasa had PMed me about her question about sandwish ELISA with brain tissue homogenates.
Now I put her PM to this forum.
Hope someone who have more experience with tissue homogenates in ELISA can help her.
"Dear Minnie, excuse me for writing to your private adress, but I think maybe be you can help me with ELISA problem. 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. Probably Im doing something wrong, i dont know what kind of control can i use, if the company claims there is no croos-reactivity with other molecules how i can know if these are false results?
Thank you in advance
Natasa
I will send you my raw results"
http://www.protocol-online.org/forums/inde...msg&id=5593
No way, something else, some other type of control?
It could be heterophile antibodies (there should really be a sticky thread on them in this sub forum by now). Have you tried incubating your (homogenate) samples with whole sera from the host species from which your Abs stem from?
There could be a number of reasons for your results though. Troubleshooting makes up a large portion of ELISA work (unfortunately), so don't panic, and just tell yourself you're learning something along the way.