Isotype Controls - Announcement

At the recent ISAC XIII congress held in Quebec City in 2006, ISAC organized the first in a series of workshops focusing on Topics in Multicolor Fluorescence. The first workshop was entitled: Appropriate Use of Isotype Controls.

The workshop resulted in some lively discussions and revealed an apparent confusion about the use and interpretation of isotype antibodies. The same issue has appeared in numerous discussions on the Purdue Cytometry Mailing list. During the workshop, we concluded that an appropriate course of action would be to collect data supporting and refuting the use of isotype antibodies.

ISAC members have already received this message and many have submitted data. We would like to thank those who were quick to send us their contributions on the topic. Now, we would like to expand this opportunity to all subscribers of this mailing list.

Recently, the ISAC membership has ratified a change to the ISAC Constitution & Bylaws Education Committee creating ISAC Standing Committee for Education. The Task Force addressing the issue of the appropriate use of isotype controls will continue as a subcommittee under this new structure.

 The goals of the subcommittee’s first task are:

• Gather and consolidate information about use of isotype antibodies
• Determine what further laboratory experiments (if any) need to be performed to clarify existing data
• Facilitate and develop a consensus guideline for appropriate use of isotype antibodies
• Educate scientific community (users, editorial boards, etc)

In our efforts to meet these goals, we ask for your help in the form of data (complete with experimental notes) in support of a particular strategy for appropriate use of isotype antibodies. The deadline for submission of contributions is June 30, 2007.

We will compile, evaluate, and organize this data in an effort to create a useful guideline for appropriate use of isotype antibodies. All contributors of published data will be acknowledged in the final manuscript. The data must not hold previous copyright.

Please upload your data to ftp://ftp.isac-net.org (you must copy and paste this address into your browser, enter Isotypes (case sensitive) for login name and data2008 for password) along with the following details:

• Sample description
• Cell source
• Fresh /Culture/Transplant
• Stimulation conditions
• Staining procedure
• Surface/Intracellular
• Dye Uptake
• Panel information
• All fluorochrome labels
• Cytometer type
• Modifications/Enhancements
• Laser lines

The committee may request to have access to the raw data (.fcs). Your permission to print will be obtained prior to publication.

We would enjoy your participation in this request for data to begin our endeavor to define background fluorescence and develop a series of ISAC guidelines for use of appropriate controls in flow cytometry.

Best regards,

Lori Krueger (Chair)
Ruud Hulspas
Lora Barsky
Mary Price
email: IsotypesEd@ISAC-net.org