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Human Antigen Processing and Presentation of Endogenous Antigen

$260
100 µl
APPLICATIONS
REACTIVITY
Human, Monkey

Application Methods: Flow Cytometry, Immunofluorescence (Immunocytochemistry), Immunohistochemistry (Paraffin), Western Blotting

Background: β2-microglobulin (B2M) is a principal component of the Major Histocompatibility Complex (MHC) class I molecule, a ternary membrane protein complex that displays fragments derived from proteolyzed cytosolic proteins on the surface of cells for recognition by the surveillance immune system (1,2). As an integral component of the MHC class I complex, β2-microglobulin plays a critically important role in immune system function (3). It has important relevance to cancer biology research; for example, research studies have shown that nearly one-third of diffuse large B cell lymphomas contain mutations that inactivate β2-microglobulin gene function, thereby allowing tumor cells to escape immune detection (4). In addition, β2-microglobulin has been identified as an amyloid preprotein with collagen-binding affinity (5); its accumulation in osteoarthritic lesions of long-term dialysis patients is reportedly a contributing factor to the condition known as amyloid osteoarthropathy (6).

$305
50 tests
100 µl
This Cell Signaling Technology antibody is conjugated to phycoerythrin (PE) and tested in-house for direct flow cytometry analysis in human cells. The antibody is expected to exhibit the same species cross-reactivity as the unconjugated β2-microglobulin (D8P1H) Rabbit mAb #12851.
APPLICATIONS
REACTIVITY
Human, Monkey

Application Methods: Flow Cytometry

Background: β2-microglobulin (B2M) is a principal component of the Major Histocompatibility Complex (MHC) class I molecule, a ternary membrane protein complex that displays fragments derived from proteolyzed cytosolic proteins on the surface of cells for recognition by the surveillance immune system (1,2). As an integral component of the MHC class I complex, β2-microglobulin plays a critically important role in immune system function (3). It has important relevance to cancer biology research; for example, research studies have shown that nearly one-third of diffuse large B cell lymphomas contain mutations that inactivate β2-microglobulin gene function, thereby allowing tumor cells to escape immune detection (4). In addition, β2-microglobulin has been identified as an amyloid preprotein with collagen-binding affinity (5); its accumulation in osteoarthritic lesions of long-term dialysis patients is reportedly a contributing factor to the condition known as amyloid osteoarthropathy (6).

$260
100 µl
APPLICATIONS
REACTIVITY
Human, Mouse

Application Methods: Western Blotting

Background: CD8+ cytotoxic T cells recognize peptides presented by MHC class I molecules on the surface of infected cells and tumor cells. The transporters associated with antigen processing 1 and 2 (TAP1 and TAP2) form the TAP complex which resides on the ER membrane and transports peptides from the cytoplasm into the ER for loading onto MHC class I molecules (1-8). In addition, TAP localized to endosomal membranes is important for cross-presentation by dendritic cells (9,10). IFN-γ produced by T cells and NK cells in response to infection causes upregulation of TAP1 and TAP2, resulting in increased antigen presentation to T cells (11). Some viral proteins inhibit TAP function or downregulate TAP expression resulting in viral immune evasion (12,13). In addition, investigators have observed reduced TAP expression in a variety of tumor types, and it is thought to be one mechanism for tumor immune evasion (14).

$115
20 µl
$269
100 µl
APPLICATIONS
REACTIVITY
Human

Application Methods: Flow Cytometry, Immunofluorescence (Immunocytochemistry), Immunohistochemistry (Paraffin), Western Blotting

Background: CD74, which is also known as the MHC Class II-associated invariant chain (Ii), is a type II transmembrane glycoprotein that plays a critical role in the antigen presentation process as a chaperone of MHC Class II proteins. It is expressed at high levels on B cells and to a lesser extent on numerous antigen presenting cell (APC) types including dendritic cells, Langerhans cells, monocytes, and macrophages as well as non-traditional APCs such as epithelial cells (1,2). CD74 was initially identified for its ability to regulate the folding and intracellular trafficking of newly synthesized MHC Class II molecules. Following expression, CD74 self-assembles as a trimer that serves as a scaffold for the assembly of MHC Class II molecules. Through this interaction, CD74 blocks the peptide binding cleft of MHC Class II molecules and prevents their premature association with endogenous polypeptides (3). Binding to CD74 also facilitates the translocation of MHC Class II molecules from the endoplasmic reticulum to the endocytic compartments during antigen presentation (4). In addition to its role as an MHC Class II chaperone, CD74 is also the receptor for macrophage migration-inhibitory factor (MIF). Binding to CD74 and its co-receptor, CD44, has been shown to induce the activation of the NFkB and ERK pathways to promote cell proliferation and survival signals (5,6). Recent studies have identified CXCR2 and CXCR4 as co-receptors for CD74 where MIF binding to CD74 complexes contributes to MIF-mediated monocyte chemotaxis and the induction of Akt signaling, respectively (7,8). Increased CD74 surface expression has been reported under inflammatory conditions and in certain types of cancer cells implying a potential role in tumorigenesis (9).

$108
250 PCR reactions
500 µl
SimpleChIP® Human TAP1 Promoter Primers contain a mix of forward and reverse PCR primers that are specific to a region of the human TAP1 promoter. These primers can be used to amplify DNA that has been isolated using chromatin immunoprecipitation (ChIP). Primers have been optimized for use in SYBR® Green quantitative real-time PCR and have been tested in conjunction with SimpleChIP® Enzymatic Chromatin IP Kits #9002 and #9003 and ChIP-validated antibodies from Cell Signaling Technology®. The peptide transporter TAP1 gene is regulated by Stat proteins in response to interferon signaling.
REACTIVITY
Human

Background: The chromatin immunoprecipitation (ChIP) assay is a powerful and versatile technique used for probing protein-DNA interactions within the natural chromatin context of the cell (1,2). This assay can be used to either identify multiple proteins associated with a specific region of the genome or to identify the many regions of the genome bound by a particular protein (3-6). ChIP can be used to determine the specific order of recruitment of various proteins to a gene promoter or to "measure" the relative amount of a particular histone modification across an entire gene locus (3,4). In addition to histone proteins, the ChIP assay can be used to analyze binding of transcription factors and co-factors, DNA replication factors, and DNA repair proteins. When performing the ChIP assay, cells are first fixed with formaldehyde, a reversible protein-DNA cross-linking agent that "preserves" the protein-DNA interactions occurring in the cell (1,2). Cells are lysed and chromatin is harvested and fragmented using either sonication or enzymatic digestion. Fragmented chromatin is then immunoprecipitated with antibodies specific to a particular protein or histone modification. Any DNA sequences that are associated with the protein or histone modification of interest will co-precipitate as part of the cross-linked chromatin complex and the relative amount of that DNA sequence will be enriched by the immunoselection process. After immunoprecipitation, the protein-DNA cross-links are reversed and the DNA is purified. Standard PCR or quantitative real-time PCR are often used to measure the amount of enrichment of a particular DNA sequence by a protein-specific immunoprecipitation (1,2). Alternatively, the ChIP assay can be combined with genomic tiling micro-array (ChIP on chip) techniques, high throughput sequencing (ChIP-Seq), or cloning strategies, all of which allow for genome-wide analysis of protein-DNA interactions and histone modifications (5-8). SimpleChIP® primers have been optimized for amplification of ChIP-isolated DNA using real-time quantitative PCR and provide important positive and negative controls that can be used to confirm a successful ChIP experiment.

$260
100 µl
APPLICATIONS
REACTIVITY
Human

Application Methods: Immunoprecipitation, Western Blotting

Background: CD8+ cytotoxic T cells recognize peptides presented by MHC class I molecules on the surface of infected cells and tumor cells. The transporters associated with antigen processing 1 and 2 (TAP1 and TAP2) form the TAP complex which resides on the ER membrane and transports peptides from the cytoplasm into the ER for loading onto MHC class I molecules (1-8). In addition, TAP localized to endosomal membranes is important for cross-presentation by dendritic cells (9,10). IFN-γ produced by T cells and NK cells in response to infection causes upregulation of TAP1 and TAP2, resulting in increased antigen presentation to T cells (11). Some viral proteins inhibit TAP function or downregulate TAP expression resulting in viral immune evasion (12,13). In addition, investigators have observed reduced TAP expression in a variety of tumor types, and it is thought to be one mechanism for tumor immune evasion (14).