I've been collecting hemolymph from bees. I put 5uL of perchloric acid in the tube where the hemolymph would be collected before the centrifugation... after that I immediately store it at -80ºC. Should this be fine or not?! I just want to make sure I am doing it properly so later I have samples to analyze.
By the way, how quick would the hemolymph start to degrade after collection, even if kept in ice during the procedure?
Any help would be greatly appreciated!
Thanks.
Hello Raquel do you speak spanish? Because of your name i imagine that....
I worked a few years ago with extractions of bees hemolymph. Please answer me about your language for a better comunication. Soledad
Hola Soledad, gracias por su respuesta.
I do speak portuguese, but I can understand castellano (written and spoken) but I am not very good at writing. So, you can answer me in castellano if you would like to. I am very much appreciated. Raquel
Bien Raquel escribir en ingles me lleva mas tiempo y ultimamente no tengo mucho disponible. Te envio un informe que realice sobre extraccion de hemolinfa en Apis mellifera para cuantificar proteinas. Puedes ver el punto 2 y 3 del escrito para que sepas la tecnica que utilice y quizas te sirva. Cualquier cosa volvemos a comunicarnos. (perdon pero mi teclado no me permite acentuar las palabras como corresponde). Saludos. Soledad
Hi Raquel, the question is what do you want to do with the hemolymph? I would recommend to put it w fat w possible at -80.
hi ~~I have the same questions!!!
I collect hemolymph to run the SDS page , after SDS page I use the coomassie blue!!
Is that appropriate ?
Hello! Thank you all for your kind replies!
1) Madanagopal Nalini, do you have any paper with that protocol you've mentioned?
2) Rolf Ziegler, that's an appropriate question indeed. For now, I intend to estimate the metal content in the hemolymph of bees using ICP-OES. I may try to estimate JH contents as well. So far I tried to collected the hemolymph directly from head and thorax by briefly centrifuging each bee upside-down in a microtube set-up (antennae and wings cut, ab removed). At the bottom of each microtube I had previously added 5uL of 0.1M perchloric acid. I was able to collect aprox 15/20uL in total per bee. Then stored at -80'C.
3) Chia YING Yen, may I ask how did that work out for you?
Thank you. BW
Soledad Garcia, which temperature did you freeze the HL (hemolymph)?
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