Western Blot shows lysates of MDA-MB-231 human breast cancer cell line, Raji human Burkitt's lymphoma cell line and THP-1 human acute monocytic leukemia cell line. PVDF membrane was probed with 2 µg/ml of Mouse Anti-Human HLA Class I Monoclonal Antibody (Catalog # MAB11502) followed by HRP-conjugated Anti-Mouse IgG Secondary Antibody (Catalog # HAF018). A specific band was detected for HLA Class I at approximately 48 kDa (as indicated). This experiment was conducted under reducing conditions and using Western Blot Buffer Group 1.

Detection of HLA Class I in Human Spleen.

HLA Class I was detected in immersion fixed paraffin-embedded sections of human spleen using Mouse Anti-Human HLA Class I Monoclonal Antibody (Catalog # MAB11502) at 1.7 µg/ml for 1 hour at room temperature followed by incubation with the HRP-conjugated Anti-Mouse IgG Secondary Antibody (Catalog # HAF007) or the Anti-Mouse IgG VisUCyte™ HRP Polymer Antibody (Catalog # VC001). Before incubation with the primary antibody, tissue was subjected to heat-induced epitope retrieval using VisUCyte Antigen Retrieval Reagent-Basic (Catalog # VCTS021). Tissue was stained using DAB (brown) and counterstained with hematoxylin (blue). Specific staining was localized to endothelial cells. View our protocol for Chromogenic IHC Staining of Paraffin-embedded Tissue Sections.

Detection of HLA Class I in PBMC by Flow Cytometry

PBMC were stained with Mouse Anti-Human CD19 APC-conjugated Monoclonal Antibody (Catalog # FAB4867A) and either (A) Mouse Anti-Human HLA Class I Monoclonal Antibody (Catalog # MAB11502) or (B) isotype control antibody (Catalog # MAB004) followed by Phycoerythrin-conjugated Anti-Mouse IgG Secondary Antibody (Catalog # F0102B). View our protocol for Staining Membrane-associated Proteins.

Applications for Human HLA Class I Antibody

Application

Recommended Usage

Flow Cytometry

2.5 µg/10⁶ cells
Sample: Detection of HLA Class I in PBMC by Flow Cytometry

Immunohistochemistry

3-25 µg/mL
Sample: Immersion fixed paraffin-embedded sections of human spleen

Western Blot

2 µg/mL
Sample: MDA-MB-231 human breast cancer cell line, Raji human Burkitt's lymphoma cell line and THP-1 human acute monocytic leukemia cell line

Formulation, Preparation, and Storage

Purification

Protein A or G purified from hybridoma culture supernatant

Reconstitution

Reconstitute lyophilized material at 0.2mg/ml in sterile PBS. For liquid material, refer to CoA for concentration.

Formulation

Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. See Certificate of Analysis for details.
*Small pack size (-SP) is supplied either lyophilized or as a 0.2 μm filtered solution in PBS.

Shipping

Lyophilized product is shipped at ambient temperature. Liquid small pack size (-SP) is shipped with polar packs. Upon receipt, store immediately at the temperature recommended below.

Stability & Storage

Use a manual defrost freezer and avoid repeated freeze-thaw cycles.

12 months from date of receipt, -20 to -70 °C as supplied.
1 month, 2 to 8 °C under sterile conditions after reconstitution.
6 months, -20 to -70 °C under sterile conditions after reconstitution.

Background: HLA Class I

HLA-A, B, and C are approximately 45 kDa transmembrane glycoproteins in the major histocompatibility complex 1 (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 12 kDa 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.

References

Barnstable, C.J. et al. (1978) Cell 14:9.

Long Name

Class I Human Leukocyte Antigen

Alternate Names

HLA-A, HLA-ABC

Entrez Gene IDs

3105 (Human)

Gene Symbol

HLA-A

Additional HLA Class I Products

Product Documents for Human HLA Class I Antibody

Product Datasheets

Download Product Datasheets

COA

SDS

Download SDS

Product Specific Notices for Human HLA Class I Antibody

For research use only

Citations
Reviews

Calculators
Flow Cytometry