Catalogue

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.

Source
HC10 is a mouse monoclonal IgG2a 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.

Product
Each vial contains 100 µl 1 mg/ml purified monoclonal antibody in PBS containing 0.09% sodium azide.

Formulation: Each vial contains 100 µl 1 mg/ml purified monoclonal antibody in PBS containing 0.09% sodium azide.

Specificity
The mouse monoclonal antibody HC10 recognizes HLA class I heavy chains. HC10 reacts mostly with HLA-B and HLA-C heavy chains and some HLA-A (HLA-A10, HLA-A28, HLA-A29, HLA-A30, HLA-A31, HLA-A32, HLA-A33).

Applications
HC10 was raised against free class I heavy chains of HLA antigens 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. HC10 indeed retains strong reactivity with free class I heavy chains in Western blots. HC10 also produces strong reactivity in immuno-electron microscopy. Its use allows the determination of tissue and subcellular distribution of class I antigen and can be used to specifically block free heavy chains (FHC); See reference 6. 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.

Storage
The antibody is shipped at ambient temperature and may be stored at +4°C. For prolonged storage prepare appropriate aliquots and store at or below -20°C. Prior to use, an aliquot is thawed slowly in the dark at ambient temperature, spun down again and used to prepare working dilutions by adding sterile phosphate buffered saline (PBS, pH 7.2). Repeated thawing and freezing should be avoided. Working dilutions should be stored at +4°C, not refrozen, and preferably used the same day. If a slight precipitation occurs upon storage, this should be removed by centrifugation. It will not affect the performance or the concentration of the product.

Shipping Conditions: Ship at ambient temperatures.

Caution
This product is intended FOR RESEARCH USE ONLY, and FOR TESTS IN VITRO, not for use in diagnostic or therapeutic procedures involving humans or animals. It may contain hazardous ingredients. Please refer to the Safety Data Sheets (SDS) for additional information and proper handling procedures. Dispose product remainders according to local regulations.This datasheet is as accurate as reasonably achievable, but our company accepts no liability for any inaccuracies or omissions in this information.

References
1. Stam NJ, Vroom TM, Peters PJ, Pastoors EB, Ploegh HL. HLA-A- and HLA-B-specific monoclonal antibodies reactive with free heavy chains in western blots, in formalin-fixed, paraffin-embedded tissue sections and in cryo-immuno-electron microscopy. Int. Immunol. 1990;2:113-125. 2. de Kruijf EM, van Nes JGH, Sajet A, Tummers QRJG, Putter H, Osanto S, Speetjens FM, Smit VTHBM, Liefers GJ, van de Velde CJH, Kuppen PJK. The predictive value of HLA Class I tumor cell expression and presence of intratumoral Tregs for chemotherapy in patients with early breast cancer. Clin Cancer Res 2010;16:1272-1280. 3. Gauster M, Blashitz A, Dohr G. Monoclonal antibody HC10 does not bind to HLA-G. Rheumatology 2007; 46: 892-893. 4. Perosa F, Luccarelli G, Prete M, Favoino E, Ferrone S, Dammacco F. Beta-2-Microglobulin-free HLA Class I heavy chain epitope mimicry by monoclonal antibody HC-10-specific peptide. J Immunol 2003;171:1918-1926. 5. Hutter H, Hammer A, Blaschitz A, Hartmann M, Ebbesen P, Dohr G, Ziegler A, Uchanska-Ziegler B. Expression of HLA class I molecules in human first trimester and term placenta trophoblast. Cell Tissue Res 1996; 286:439–447. 6. Abdullah H, Zhang Z, Yee K, Haroon N. KIR3DL1 interaction with HLA-B27 is altered by ankylosing spondylitis associated ERAP1 and enhanced by MHC class I cross-linking. Discov Med. 2015;20(108):79-89 7. Goodridge JP, Burian A, Lee N, Geraghty DE. HLA-F and MHC class I open conformers are ligands for NK cell Ig-like receptors.J Immunol. 2013;191(7):3553-62

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