Mouse Anti-HLA Class 1 Antigen ABC (HLA ABC, Major Histocompatibility Complex Class I A + B + C, MHC HLA ABC) (PerCP)
HLA-A, B, and C are ~45kD transmembrane glycoproteins in the major histocompatibility complex I (MHC I) family. They contain three alpha domains in their extracellular regions. HLA molecules are expressed on nearly all nucleated cells in association with the 12kD beta 2-Microglobulin. This complex binds peptides derived from pathogenic cytosolic or extracellular proteins such as viral or microbial proteins. It presents these peptides on the cell surface for recognition by the T cell receptor on CD8+ cytotoxic T cells. The activated cytotoxic T cell then kills the presenting cell. Mismatched MHC I alleles between a host and a donor lead to transplant rejection.
Applications
Suitable for use in Flow Cytometry. Other applications not tested.
Recommended Dilution
Flow Cytometry: 10ul labels 10e6 cells in human peripheral blood lymphocytes Optimal dilutions to be determined by the researcher.
Flow Cytometry Protocol
1. Cells may be Fc-blocked with 1ug of human IgG/10e5 cells for 15 minutes at RT. Do not wash excess blocking IgG from this reaction. 2. After blocking, 10ul of conjugated antibody was added to 1- 2.5x10e5 cells and incubated for 30 minutes at RT. 3. Unbound antibody was removed by washing the cells twice in Flow Cytometry Staining Buffer (Catalog # F4568-10). Note that whole blood requires a RBC lysis step at this point using Flow Cytometry Human Lyse Buffer. 4. Resuspend the cells in Flow Cytometry Staining Buffer for final flow cytometric analysis. As a control for this analysis, cells in a separate tube should be treated with PerCP-labeled mouse IgG2A antibody.
Storage and Stability
May be stored at 4°C. For long-term storage, aliquot and store at 4°C. Do not freeze. Aliquots are stable for 12 months after receipt. For maximum recovery of product, centrifuge the original vial prior to removing the cap. Further dilutions can be made in assay buffer. PerCP conjugates are sensitive to light.
Immunogen
Membranes from human tonsillar lymphocytes
Form
Supplied as a liquid in saline, 0.5% BSA, 0.09% sodium azide. Labeled with PerCP.
Purity
Purified by Protein A affinity chromatography from culture supernatant.
Specificity
Recognizes human MHC class I, HLA-A, B, and C. Recognizes a non-polymorphic epitope shared among products of the HLA-A, B, and C loci and immunoprecipitates both the HLA molecule and beta 2-Microglobulin.
