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youyou's Content

There have been 8 items by youyou (Search limited from 13-August 19)


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#29685 Inducing Adherence in Suspension Cells

Posted by youyou on 20 July 2009 - 06:30 AM in Tissue and Cell Culture

Hello all,

I'm looking for an easy way to induce adherence of a Jurkat suspension cell line I'm currently running. Some protocols I've found have suggested incubation in serum-free media (~ 10 mins in a collagen-coated plasticware) prior to doing fixation, but I just wanted to get some thoughts. I need to find an easier way to induce adherence in these cells for translocation assays, and spinning everytime I need to do a wash step is time-consuming and tedious. Any thoughts are appreciated!


Hi

could you please pass me some of the protocols or articles you have found ? i would be grateful , and have you tried using fibronectin or vitronectin coating of plasticware to adhere the suspension cells ? but attention it also means incubation in serum free medium .



#29684 suspension cell culture

Posted by youyou on 20 July 2009 - 06:22 AM in Tissue and Cell Culture

Hi everyone

i have a question about the optimum number of cells in suspension that can be cultured in multi well plate i.e in a 24 well plate what is the max. no. of cells that can be cultured for a period of 72 hours???????

thanx for your opinions in advance

yusra



#24727 RT-PCR

Posted by youyou on 20 May 2009 - 12:23 AM in PCR, RT-PCR and Real-Time PCR

Hi everyone

I have a small oligo primer of 20 mer and the PCR product size should be at 158bp and i am only getting bands at 1.5kb band on gel electrophoresis which is too large!! My oligo primers (F and R) have the same melting temperature at 62'C so i used an annealing temperature equal to the melting temperature for 28 cycles. Can anyone help me on how to calculate the required temperatures and how to programme my elongation?????????????

Can you please send me a link to help me with PCR ??????????

thanx in advance.



#22256 coating tissue culture lates with fibronectin

Posted by youyou on 20 April 2009 - 06:14 AM in Tissue and Cell Culture

Hi everyone

could someone advice me on how to coat my tissue culture plates with fibronectin? the negative and positive controls? and do i have to culture my cells in serum free medium in this condition?
Thanx



#21493 protein array

Posted by youyou on 10 April 2009 - 06:51 AM in Macroarray and Microarray

hi evryone


i would like to ask if the protein micro array should be done for cells that have been cultured in serum free culture media.
And if not would this mean the presence of a background fond.

thanx



#20795 Trypan Blue and Hoechst staining..

Posted by youyou on 02 April 2009 - 07:14 AM in General Lab Techniques

HI


I saw your question, I think it would be better if you used the annexin or the propidium iodide which are more precise for the detection of cell mortality.



#19104 what is meant by passages

Posted by youyou on 17 March 2009 - 09:07 AM in Tissue and Cell Culture

Hi

i wanted to know exactly what is meant by passage in cell culture and how many passages can be max. done for Nalm6 cells ? :(



#19103 Cell count using a hemocytometer

Posted by youyou on 17 March 2009 - 09:00 AM in Tissue and Cell Culture

This is different from the mallasez counting chamber , since the size of each square is 1mm .




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