Anti-Human HLA-A, B, C (MHC Class I) [Clone W6/32] — Purified in vivo PLATINUM™ Functional Grade
Anti-Human HLA-A, B, C (MHC Class I) [Clone W6/32] — Purified in vivo PLATINUM™ Functional Grade
Product No.: H463
Clone W6/32 Target HLA-A,B,C Formats AvailableView All Product Type Monoclonal Antibody Alternate Names Major Histocompatibility Class I, MHC class I, human leukocyte antigen (HLA) Isotype Mouse IgG2a k Applications B , FC , IHC FF , in vivo , IP , PhenoCycler® , WB |
Antibody DetailsProduct DetailsReactive Species Baboon ⋅ Chimpanzee ⋅ Cynomolgus Monkey ⋅ Feline ⋅ Bovine ⋅ Human Host Species Mouse Recommended Isotype Controls Recommended Isotype Controls Recommended Dilution Buffer Immunogen Human tonsil cell membrane Product Concentration ≥ 5.0 mg/ml Endotoxin Level ≤ 0.5 EU/mg as determined by the LAL method Purity ≥98% monomer by analytical SEC ⋅ >98% by SDS Page Formulation This monoclonal antibody is aseptically packaged and formulated in 0.01 M phosphate buffered saline (150 mM NaCl) PBS pH 7.2 - 7.4 with no carrier protein, potassium, calcium or preservatives added. Due to inherent biochemical properties of antibodies, certain products may be prone to precipitation over time. Precipitation may be removed by aseptic centrifugation and/or filtration. Product Preparation Functional grade preclinical antibodies are manufactured in an animal free facility using in vitro cell culture techniques and are purified by a multi-step process including the use of protein A or G to assure extremely low levels of endotoxins, leachable protein A or aggregates. Pathogen Testing To protect mouse colonies from infection by pathogens and to assure that experimental preclinical data is not affected by such pathogens, all of Leinco’s Purified Functional PLATINUM<sup>TM</sup> antibodies are tested and guaranteed to be negative for all pathogens in the IDEXX IMPACT I Mouse Profile. Storage and Handling Functional grade preclinical antibodies may be stored sterile as received at 2-8°C for up to one month. For longer term storage, aseptically aliquot in working volumes without diluting and store at ≤ -70°C. Avoid Repeated Freeze Thaw Cycles. Country of Origin USA Shipping Next Day 2-8°C RRIDAB_2893098 Applications and Recommended Usage? Quality Tested by Leinco FC The suggested concentration for this HLA-A,B,C Clone W6/32 antibody for staining cells in flow cytometry is ≤ 2.0 μg per 106 cells in a volume of 100 μl or 100μl of whole blood. Titration of the reagent is recommended for optimal performance for each application.
WB The suggested concentration for for use in western blotting is 1-10 μg/ml. WB Bit1 antibody can be used for the detection of Bit1 by Western blot at 1 - 4 μg/mL. Additional Applications Reported In Literature ? B PhenoCycler-Fusion (CODEX)® IHC FF IP Each investigator should determine their own optimal working dilution for specific applications. See directions on lot specific datasheets, as information may periodically change. DescriptionDescriptionSpecificity Clone W6/32 recognizes the human MHC class I molecules HLA-A, -B, and -C.
Background HLA antibody, clone W6/32, recognizes the major histocompatibility complex (MHC) class I molecules human leukocyte antigen (HLA)-A, HLA-B, and HLA-C. MHC class I is ubiquitously expressed on the cell surface of nucleated cells and consists of a 45-kDa type I transmembrane glycoprotein (α-chain or heavy chain) and a 12-kDa soluble protein (β2-microglobulin, β2M)1,2. The α-chain consists of three domains (α1, α2, and α3)3. α1 and α2 form the closed antigen-binding groove and bind to 8-10 aa peptides derived from cytosolic antigens4-6. β2M noncovalently associates with α3, which is essential for MHC stability. MHC class I plays a critical role in the adaptive immune response by presenting endogenous antigens to cytotoxic CD8 T cells. MHC class I molecules can also present exogenous antigens to CD8 T cells via a process known as cross-presentation7. The T cell receptor (TCR)/CD3 complex of CD8 T cells interacts with peptide-MHC class I, which induces CD8 T cell activation and subsequent cell-killing. CD8 molecules also bind to MHC class I, which helps augment TCR signaling8. In contrast to CD8 T cells, MHC class I is an inhibitory ligand for natural killer (NK) cells, promoting self tolerance9. MHC class I also contributes to the positive selection of CD8 T cells and NK cell specificity10,11.
Antigen Distribution HLA-A, -B, and -C are ubiquitously expressed on nucleated cells.
Ligand/Receptor CD3/TCR, CD8 Function Antigen presentation PubMed NCBI Gene Bank ID UniProt.org Research Area Immunology . Innate Immunity Leinco Antibody AdvisorPowered by AI: AI is experimental and still learning how to provide the best assistance. It may occasionally generate incorrect or incomplete responses. Please do not rely solely on its recommendations when making purchasing decisions or designing experiments. Clone W6/32 is a mouse monoclonal antibody specific for human MHC Class I molecules (HLA-A, B, and C), and in mice, its common in vivo applications mainly involve studies using humanized mouse models rather than wild-type or standard laboratory mice. Key in vivo applications of W6/32 in mice:
Important Considerations:
Summary Table: W6/32 In Vivo Use in Mice
Clone W6/32 is not functional for direct mouse MHC detection; its in vivo use in mice is exclusively tied to experimental systems incorporating human cells or tissues such as humanized mouse models, where it provides a critical tool for validating and analyzing human immune processes. Other commonly used antibodies or proteins alongside W6/32 in the literature include those that recognize different epitopes or subclasses of HLA class I, HLA class II molecules, or associate with immunological reference markers such as β2-microglobulin, CD8, and NK cell ligands. Key examples often found in the literature:
Research context also frequently determines additional marker selection:
HLA-E–specific antibodies such as 3D12 and 4D12 are also commonly cited, particularly in contexts where non-classical class I molecules are relevant. In summary, common pairs or panels with W6/32 include:
Selection depends on the experimental goal: detection, functional analysis, or specificity mapping. Clone W6/32 is a monoclonal antibody that has been extensively used in immunological research for over six decades, with several key findings emerging from its widespread application in scientific literature. Recognition of a Conserved HLA Class I EpitopeThe most fundamental discovery is that W6/32 recognizes a conformational epitope that is conserved across diverse HLA-I allotypes, effectively bypassing the extreme polymorphism that typically confounds HLA class I studies. This monomorphic epitope is expressed on native β2-microglobulin (β2m)-associated Major Histocompatibility Complex (MHC) class I molecules, specifically HLA-A, HLA-B, and HLA-C. The antibody binds to a discontinuous epitope comprised of the α1, α2, and α3 domains of the HLA-I heavy chain along with β2-microglobulin. Structural CharacterizationCrystal structure analysis has revealed that W6/32 binds beneath the antigen-binding groove of HLA class I molecules. The epitope comprises a region of low polymorphism, which explains the pan-HLA-I nature of its binding capability. This structural positioning is particularly significant because it does not overlap with binding sites for T cell antigen receptors or killer cell immunoglobulin-like receptors (KIRs). Functional Implications for Immune Cell InteractionsA critical finding is that the W6/32 epitope coincides with binding sites for leukocyte immunoglobulin-like receptors (LILRs) and CD8 coreceptors. Functional studies demonstrated that using W6/32 to block the interaction between NK cells and HLA-I only weakly impaired inhibition mediated by KIR3DL1, but significantly impacted HLA-LILR recognition. This selective blocking pattern provides important insights into differential receptor-ligand interactions on HLA class I molecules. Clinical and Research ApplicationsThe antibody has proven valuable for detecting variations in HLA class I expression across different cell types and disease states. Research has documented that HLA class I expression is upregulated on activated cells or those responding to proinflammatory cytokines, while reduced expression is found on certain virus-infected or tumor cells. Studies have identified cases where tumors show complete loss of HLA class I expression, with some pancreatic carcinomas testing negative for W6/32. The ability of W6/32 to recognize its epitope only on properly folded, β2m-associated HLA class I molecules makes it particularly useful for assessing the native conformation and stability of these proteins in various experimental contexts. Dosing regimens of clone W6/32 are not standardized across mouse models, because this monoclonal antibody is primarily used for ex vivo or in vitro applications—such as flow cytometry and immunofluorescence detection of human HLA class I molecules—rather than for systemic administration in live mice. Most suppliers and reviews confirm that no established in vivo dosing protocol exists for W6/32 in typical mouse strains, as it is not intended for immunomodulation or depletion in murine models. Key context and details:
Exception—rare in vivo use:
Summary Table: W6/32 Use Contexts
Conclusion: References & Citations1. Mitaksov V & Fremont DH. (2006) J Biol Chem. 281(15):10618-25 2. Wieczorek M, et al. (2017) Front Immunol. 8:292 3. Jones EY. (1997) Curr Opin Immunol. 9(1):75-9 4. Matsumura M, et al. (1992) Science. 257:927–34.10.1126/science.1323878 5. Bouvier M & Wiley DC. (1994) Science. 265:398–402.10.1126/science.8023162 6. Zacharias M & Springer S. (2004) Biophys J. 87:2203–14.10.1529/biophysj.104.044743 7. Cruz FM, et al (2017) Annu Rev Immunol. 35:149-176 8. Artyomov MN, et al (2010) Proc Natl Acad Sci USA. 107(39):16916-16921 9. Orr MT & Lanier LL. (2010) Cell. 142(6):847-856 10. Raulet DH. (1994) Adv Immunol. 55:381-421 11. Salcedo M & Ljunggren HG. (1996) Chem Immunol. 64:44-58 Technical ProtocolsCertificate of Analysis |
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