Anti-Human HLA-A, B, C (MHC Class I) - Purified In vivo PLATINUM Functional Grade

Antibody Details

Product Details

Reactivity Species

Baboon

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Chimpanzee

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Cynomolgus Monkey

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Feline

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Bovine

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Human

Host Species

Mouse

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

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> 95% 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.

Product Preparation

Functional grade preclinical antibodies are manufactured in an animal free facility using only In vitro protein free 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^TM antibodies are tested and guaranteed to be negative for all pathogens in the IDEXX IMPACT I Mouse Profile.

Storage and Handling

This antibody is stable for at least one week when stored at 2-8°C. For long term storage, aliquot in working volumes without diluting and store at – 20°C in a manual defrost freezer. Avoid Repeated Freeze Thaw Cycles.

Country of Origin

USA

Shipping

2-8°C

RRID

AB_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.

Other Applications Reported In Literature ?

B
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.

Description

Specificity

Clone W6/32 recognizes the human MHC class I molecules HLA-A, -B, and -C.

Antigen Distribution

HLA-A, -B, and -C are ubiquitously expressed on nucleated cells.

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 antigens^4-6. 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 specificity^{10, 11}.

Antigen Details

Protein

HLA-A, B, C

Ligand/Receptor

CD3/TCR, CD8

Function

Antigen presentation

PubMed

HLA-A, B, C

NCBI Gene Bank ID

3105

UniProt.org

Information on Uniprot.org

Research Area

Immunology

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Innate Immunity

References & Citations

1. 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 Protocols