Mouse C57BL Splenocytes
≥ 5.0 mg/ml
<1.0 EU/µg as determined by the LAL method
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.
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.
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
Next Day 2-8°C
Applications and Recommended Usage?
Quality Tested by Leinco
FC The suggested concentration for clone AF6-88.5 antibody for staining cells in flow cytometry is ≤ 0.25 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.
Other Applications Reported In Literature ?
Each investigator should determine their own optimal working dilution for specific applications. See directions on lot specific datasheets, as information may periodically change.
Clone AF6-88.5, Anti-Mouse H-2Kb, recognizes an epitope on mouse MHC class I H-2K haplotype b. AF6-88.5 does not cross-react with other H-2K haplotypes, including d, f, j, k, p, q, r, s, u, v.
The AF6-88.5 antibody reacts with H-2Kb MHC class I alloantigen and is ubiquitously expressed on nucleated cells from mice of the H-2Kb haplotype, including C57BL/6. H-2Kb is involved in antigen presentation to T cells expressing CD3/TCR and CD8 proteins.
H-2Kb antibody, clone A4C8.1-Do9, recognizes the major histocompatibility complex (MHC) class I H-2K haplotype b (H-2Kb). 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. H-2Kb 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 to 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.
NCBI Gene Bank ID
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
Products are for research use only. Not for use in diagnostic or therapeutic procedures.