Hi Attila, Do you mean you wish to perform a HLA Class II type on these cells? if so you are looking at the wrong technology, HLA typing (especially class II) is now usually performed using molecular methods either PCR-SSP or PCR-SSO. If you are new to this you should buy a commercial kit (try Dynal at http://www.tissue-typing.com) Only immunologically active cells express Class II so B Cells and activated T Cells are the usual ones to go for, Class 2 molecules are divided up into DRA,DRB1,DRB3,DRB4,DRB5, DQA,DQB, DPA,DPB as well as some minor HLA antigens (sorry no such thing as D) most of these should be present but its not the fact that there present that is important its what type are present there are many!! and for this you will need a molecular method. Good Luck John ----- Original Message ----- From: "Dr. Attila Tarnok" <tarnok@medizin.uni-leipzig.de> To: cyto-inbox Sent: Wednesday, November 15, 2000 4:14 PM Subject: MHC classII typing > > Dear Colleaugues, > > we want to perform MHC II typing from peripheral blood in patients with > suspected autoimmune disease. Being completely unexperienced in this field I > have some very basic questions to the community. > > 1. Is it sufficient to stain the cells for HLA-DP, HLA-DQ, HLA-DR (I only found > ABs to these subclasses) or do I need more antibodies? > 2. Is it ueseful to use additionally an AB against the common HLA-D molecule? > 3. Can I do all stainings in one well and use directly labelled ABs? > 4. Where can I get HLA-DP directly dye labelled? > 5. Should I gate on B-cells or monocytes? Does this matter at all? > 6. What results should I expect: Is a patient either one of the HLA-D types or > do I get a mix of all three types? > 7. Anything important that should be taken into account? > > Any help is appreciated. Thanks > Attila Tarnok > > >
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