>On Mon, 9 Mar 1998, frederic clement wrote:
>>> Dear scientists,
>>>> Can anyone help me with commercial available permeabilisation kits or
>> other commercial stuff for detection of intracellular cytokines by
>> flow-cytometry ?
>>>> Thanks a lot.
>>>> PS : Please mail me your advice.
>>>>>Several companies including Pharmingen (San Deigo) and Coulter sell kits
>to permeabilize cells. However, inour laboratory we have found that the
>following procedure works quite well.
>>1) Surface stain as normal
>2) fix with 4%formaldehyde
>3) Wash with PBS
>4) add PBS containing 0.1% saponin and stain intracellularly for 30 min.
>5) Wash out intracellular stain with PBS-azide containing 0.1% saponin.
>6) fix with 1% formaldehyde
>>This is the basic procedure. The literature has published several methods
>based on this type of procedure
>>Good luck
>>Dr. Tim Wyant
>>Also, an Appendix in a Pharnimgen catalogue describes a intracellular
staining procedures. Many of the commercially available kits nowadays
come with Brefeldin A and/or Monesin which may or may not be effetive.
regards,
Aki Hoji (akhst7+ at pitt.edu)
Dept. Infectious Diseases and Microbiology
University of Pittsburgh
