BD fastimmune protocol
CPRUSSIN at atlas.niaid.nih.gov
Wed Feb 26 19:41:13 EST 1997
I am concerned that by only collecting 3000 CD3+ cells you will be
generating less than the most robust numbers for IL-4 frequencies. The
significance of a frequency is proportional to the number of events
you acquire. David Parks has a table in the FACS chapter he wrote for
Paul's Fundamental Immunology, 2nd edition. By only collecting 3000
events, a value of 5% could be anything from 4 to 6%. By increasing
that to 50,000 events, you narrow the range to 4.7% to 5.3%.
Laboratory of Allergic Diseases
NIAID/ National Institutes of Health
From: Michelle N Fiordalisi
Sent: Wednesday, February 26, 1997 9:39 AM
Subject: Re: BD fastimmune protocol
Since I've had several requests for a workable protocol for
cytoking staining, I'd lot I'd post it for every one.
The procedure I've been using is based on the BD protocol published in
their Application Note 1.
I then acquire 3000-5000 events in a CD3+ gate and analyze by
quadrant analysis (CellQuest) of dot plots. I calculate % positivity
using BD's formula: (Activ. sample - activ. isotype control) - (
sample - Unstim. isotype control).
Michelle N. Fiordalisi, Ph.D.
Clinical Immunology Fellow
University of North Carolina Hospitals
email: mnfiord at med.unc.edu
Phone: 919-966-4058 FAX: 919-966-0486
More information about the Cytometry