HLA Class I Heavy Chain antibody

Catalog No. ABIN5691101

Product Details

Target: HLA Class I Heavy Chain

Reactivity: Human

Host: Mouse

Clonality: Monoclonal

Conjugate: Un-conjugated

Application: ELISA, Western Blotting (WB), Flow Cytometry (FACS), Immunoprecipitation (IP), Electron Microscopy (EM), Immunocytochemistry (ICC), Immunohistochemistry (Frozen Sections) (IHC (fro)), Immunohistochemistry (Paraffin-embedded Sections) (IHC (p))

Specificity: The mouse monoclonal antibody HCA2 recognize HLA class I heavy chains. The reactivity spectrum of HCA2 is composed of all HLA-A chains (except HLA-A24), as well as some HLA-B, HLA-C, HLA-E, HLA-F, and HLA-G chains.

Immunogen: HCA2 is a mouse monoclonal IgG1 antibody derived by fusion of SP2/0-Ag14 mouse myeloma cells with spleen cells from BALB/c mice immunized with HLA-B7 and -B40 heavy chains.

Clone: HCA2

Isotype: IgG1

Application Details

Application Notes: The antibody HCA2 reacts preferentially with HLA-A locus heavy chains. HCA2 was raised against free class I heavy chains of HLA, to obtain antibodies that would still react with denatured class I antigens, as they occur in Western blotting, conventional light microscopical analysis of formalin-fixed, paraffin-embedded sections, and cryo-immuno-electron microscopy. HCA2 indeed retains strong reactivity with free class I heavy chains in Western blots. HCA2 in particular reacts in a locus-specific manner by biochemical criteria. Conditions are described for use of HCA2 in immunohistochemical staining of formalin-fixed, paraffin-embedded sections (see references). HCA2 also produces strong reactivity in immuno-electron microscopy. Its use allows the determination of tissue and subcellular distribution of class I antigens. Optimal antibody dilutions for the different applications should be determined by titration, recommended range is 1:100 - 1:200 for flow cytometry, and for immunohistochemistry with avidin-biotinylated horseradish peroxidase complex (ABC) as detection reagent, and 1:100 - 1:1000 for immunoblotting applications.

Restrictions: For Research Use only

Handling

Buffer: Each vial contains 100 μL 1 mg/mL purified monoclonal antibody in PBS containing 0.09 % sodium azide.

Preservative: Sodium azide

Precaution of Use: This product contains Sodium azide: a POISONOUS AND HAZARDOUS SUBSTANCE which should be handled by trained staff only.

Storage: 4 °C,-20 °C

Storage Comment: Store at 4°C, or in small aliquots at -20°C.

References

Dijkstra, Cattaneo, Weeber, Chalabi, van de Haar, Fanchi, Slagter, van der Velden, Kaing, Kelderman, van Rooij, van Leerdam, Depla, Smit, Hartemink, de Groot, Wolkers, Sachs, Snaebjornsson, Monkhorst et al.: "Generation of Tumor-Reactive T Cells by Co-culture of Peripheral Blood Lymphocytes and Tumor Organoids. ..." in: Cell, Vol. 174, Issue 6, pp. 1586-1598.e12, (2019) (PubMed).

Sundara, Kostine, Cleven, Bovée, Schilham, Cleton-Jansen: "Increased PD-L1 and T-cell infiltration in the presence of HLA class I expression in metastatic high-grade osteosarcoma: a rationale for T-cell-based immunotherapy." in: Cancer immunology, immunotherapy : CII, Vol. 66, Issue 1, pp. 119-128, (2017) (PubMed).

Target Details

Target: HLA Class I Heavy Chain

Background: The HLA class I gene family is composed of a group of genes whose products encode cell surface glycoproteins of MW 40-45 kDa, associated non-covalently with the beta-2-microglobulin light chain. They include the three polymorphic molecules HLA-A, -B, and -C, which are ubiquitously expressed and which are able to present intracellular peptides to cytotoxic T cells. Three additional class I genes are known, commonly referred to as non-classical or class Ib genes, all highly homologous to the other class I genes and all of which associate with beta-2-microglobulin light chain. In humans, each of the class Ib genes appears to exhibit a distinct pattern of expression in developing and adult tissues. HLA-E transcripts are distributed widely in adult tissues and have also been found in the placenta and fetal liver. In the adult, the presence of HLA-F has been shown in skin, resting T cells, and B cells, whereas its expression during development has been reported in fetal liver and at low levels in placenta and extra-placental tissues. HLA-G was originally thought to be expressed only in certain populations of placental trophoblasts, but low levels have also been found in a variety of human tissues. Recently it was shown that HLA class I expression in breast cancer cells can have a predictive value for chemotherapy response.