Human MHC Class II (HLA-DP, DQ & DR) molecules are expressed on dendritic cells, monocytes, macrophages, and myeloid cells. MHC Class II molecules are heterodimers of polymorphic transmembrane a and b chains and generally bind exogenously derived peptides of about 10-17aa MHC Class II molecules interact with the T cell receptor on CD4+ T cells.
Applications:
Suitable for use in Flow Cytometry and Western Blot.
Other applications not tested.
Recommended Dilutions:
Flow Cytometry: 1:50 (5ug/ml) labels 5x10e5 human Daudi cells
Optimal dilutions to be determined by the researcher.
FACS Protocol:
1. Five x10e5 cultured human Daudi cells were pre-incubated
with 20ul of human IgG at 300ug/ml (to block non-specific binding), after which they were incubated on ice for 45 minutes with 80ul of M3887-55C at a 1:50 dilution factor (5ug/ml).
2. Cells were washed three times, fixed and analyzed by FACS.
3. Cells stained positive with a mean shift of 2.1 log10 fluorescent units when compared to a Mouse IgG1/FITC negative control at a similar concentration.
4. Binding was blocked when cells were pre incubated 10 minutes with 20ul of 0.5mg/ml M3887-55A MHC Class 2 (beta chain).
Storage and Stability:
May be stored at 4 degrees C for short-term only. Aliquot to avoid repeated freezing and thawing. Store at -20 degrees C. Aliquots are stable for at least 12 months. For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap.