Fluidigm protocols?
Hi all,
I am new to the Cytof world, it is nice to have an informative and supportive community here. I will be managing a Helios instrument at JAX-GM in Farmington, CT. I am currently standardizing procedures for getting the instrument and service running and I have a couple of newbie questions:
1. How have the Fluidigm protocols worked for everybody? I have seen other protocols with custom buffers; is that purely a matter of cost reduction?
2. What is your general workflow for testing a new Ab? Buy pure--use fluorochrome secondary for flow testing-- metal conjugate?Also, has anyone encountered issues with metal conjugating polyclonal antibodies from rabbit?
3. Have you been able to successfully combine intracellular and nuclear antigen staining into the same procedure?
Thank you in advance for any input.
Sincerely,
Santhosh Sivajothi
I am new to the Cytof world, it is nice to have an informative and supportive community here. I will be managing a Helios instrument at JAX-GM in Farmington, CT. I am currently standardizing procedures for getting the instrument and service running and I have a couple of newbie questions:
1. How have the Fluidigm protocols worked for everybody? I have seen other protocols with custom buffers; is that purely a matter of cost reduction?
2. What is your general workflow for testing a new Ab? Buy pure--use fluorochrome secondary for flow testing-- metal conjugate?Also, has anyone encountered issues with metal conjugating polyclonal antibodies from rabbit?
3. Have you been able to successfully combine intracellular and nuclear antigen staining into the same procedure?
Thank you in advance for any input.
Sincerely,
Santhosh Sivajothi