Mouse HLA Class 2 Antigen DR52 Monoclonal Antibody | anti-HLA-DRB3 antibody

HLA Class 2 Antigen DR52 (R0.84)

Cat. Num. AAA603244

• Gene Names: HLA-DRB3; HLA-DR3B
• Purity: Ascites; Ascites
• Synonyms: HLA Class 2 Antigen DR52; Monoclonal Antibody; HLA Class 2 Antigen DR52 (R0.84); Anti -HLA Class 2 Antigen DR52 (R0.84); anti-HLA-DRB3 antibody

• Host: Mouse
• Clonality: Monoclonal
• Isotype: IgM
• Clone Number: 4i141
• Specificity: Reacts to HLA Class 2 Antigen-DR52. Specificity was determined at a 1:10 dilution by the microcytotoxicity test under standard NIH conditions.
• Purity/Purification: Ascites; Ascites
• Form/Format: Supplied as a lyophilized powder in PBS, 1% BSA. Reconstitute to 100ul sterile dH2O to make a 10X stock solution.

• Application Notes: Suitable for use in Flow Cytometry, Cell Typing, Tissue Staining, and Chimerism Studies.; Dilution: Cytotoxicity: Add 0.9ml of 1% BSA in PBS to 100ul of10X working dilution.
• Immunogen: HLA Class 2 Antigen-DR52.
• Cross-reactivity: Cross-reactivity with related HLA Class 2 Antigen-DR5 has been tested during the development of the MBS603244 antibody. MBS603244 has an R value of 0.84. The R value obtained from the specificity testing using the microcytoxicity test. The R-value or correlation coefficient is calculated from the following:; ; (AD) - (BC); r = ---------------------------------------------; [(A+B)(C+D)(A+C)(B+D)] 1/2; ; where; A = Number of cells with the antigen and positive with the serum; B = Number of cells with the antigen but are negative (false negative); C = Number of cells without the antigen but are positive (false positive); D = Number of cells without the antigen and negative with the serum; ; Reference for the NIH microcytotoxicity assay:; Ray, JG, Hare, DB and Pedersen PD.,eds. (1976) Manual of Tissue Typing Techniques (National Institutes of Health, Bethesda,MD), DHEW Pub.no. [NIH] 76-545.
• Protocol: INTENDED USE; For use in the detection of human leukocyte antigens (HLA) on the surface of cells. ; ; SUMMARY AND EXPLANATION; The antibodies recognize private and public determinants on HLA molecules. ; ; PRINCIPLE; These antibodies were characterized using the microcytotoxicity test. Briefly, viable lymphocytes are incubated with complement-binding antibody. If the lymphocytes express an antigen recognized by a specific antibody, the Fab portion of the antibody binds to the antigen, forming antigenantibody complex. After these complexes have formed, rabbit complement is added. The C1q and Ca++ from the complement binds to the FC portion of the antibody. One IgM antibody is required to bind one molecule of C1q, or two IgG antibodies are required to bind one molecule of C1q. Binding of C1q with antigen-antibody complexes initiates the complement cascade, which leads to cell lysis. In a negative reaction, the lymphocytes are alive. In a positive reaction, the lymphocytes are dead.. ; ; REAGENTS; 1. Identification HLA Bulk Murine Monoclonal Antibodies are supplied in lyophilized form (100ul). Specificity and dilution factors are determined by the microcytotoxicity test using NIH standard conditions. ; 2. Warning or Caution; A. For Research Use Only.; B. Caution: Avoid repeated freezing and thawing of control reagents after reconstitution. ; 3. Storage Instructions: Store at 4° C. After reconstitution, store at -20° C. Avoid high temperatures. ; 4. Purification or Treatment Required for Use Reconstitution is required for use of lyophilized reagents. ; 5. Instability Indications Do not use if antibody has not been stored properly. ; ; INSTRUMENT REQUIREMENTS; • Centrifuge; • Phase contrast microscope; ; SPECIMEN COLLECTION AND PREPARATION; Since viable lymphocytes are required for the microcytotoxicity test, blood should be received and processed immediately following procurement.; Lymphocyte yield decreases with time and extreme temperature. Blood should be collected in acid citrate dextrose (ACD) or sodium heparin and should be stored horizontally at RT (and processed within 48 hours for maximum T and B lymphocyte yield). ; ; PROCEDURE; A. Materials Provided: MBS603244 HLA Class 2 Antigen DR52 (R0.84) ; B. Materials Required, But Not Provided Sterile dH2O, PBS 10X solution, 1% BSA in PBS; ; DIRECTIONS FOR USE; 1. Preparing BSA; A. Measure out 10g of BSA (protease free). ; B. Dissolve BSA in 100ml of PBS 10X solution. ; C. Add 900ml of sterile water to make a final volume of 1 liter. ; 2. Preparing HLA Bulk Monoclonal Antibodies for Use; A. To avoid volume loss, in a microcentrifuge, centrifuge vial for a few seconds before opening. (Powder may accumulate in cap during shipment). ; B. Reconstitute with 100ul of distilled water to obtain a 10X working dilution. ; C. For a cytotoxic working dilution, add 0.9ml of 1% BSA in PBS to 100ul working dilution. ; ; LIMITATIONS OF THE PROCEDURE; ; Cell isolation difficulties and contamination of the lymphocyte preparation with red cells, monocytes, platelets, or granulocytes may cause erroneous results. In addition, erroneous results may occur when cell concentrations are above or below acceptable levels. Bacterial contamination or change in the pH of the antisera may cause false negative reactions.; ; EXPECTED VALUES; Cell death will occur in any test well in which the HLA cell surface antigen is recognized by its matched anti-HLA antibody. Live lymphocytes indicate a negative reaction. Dead lymphocytes indicate a positive reaction.
• Preparation and Storage: Lyophilized powder may be stored at 4 degree C for short-term only. Reconstitute to nominal volume by adding sterile 40-50% glycerol and store at -20 degree C. Reconstituted product is stable for 12 months at -20 degree C. For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap. Further dilutions can be made in assay buffer.; Please contact MyBioSource for bulk requirements and licensing requests relating to production applications.

Product Categories/Family for anti-HLA-DRB3 antibody

Antibodies; Abs to HLA

NCBI and Uniprot Product Information

• NCBI GI #: 4753784
• NCBI GeneID: 3125
• UniProt Accession #: Q9UQT9; P13761; P79483
• Molecular Weight: 10,806 Da
• NCBI Official Full Name: human leucocyte antigen DR52 beta 1 chain
• NCBI Official Synonym Full Names: major histocompatibility complex, class II, DR beta 3
• NCBI Official Symbol: HLA-DRB3
• NCBI Official Synonym Symbols: HLA-DR3B
• NCBI Protein Information: major histocompatibility complex, class II, DR beta 3; DR7; MHC class II antigen DRB3; human leucocyte antigen DRB3; MHC class II HLA-DR beta 3 chain; MHC class II antigen DR beta 3 chain; HLA class II histocompatibility antigen, DR beta 3 chain; HLA class II histocompatibility antigen, DRB1-7 beta chain
• UniProt Protein Name: Leucocyte antigen DR52 beta 1 chain
• Protein Family: HLA class II histocompatibility antigen
• UniProt Gene Name: HLA-DRB3
• UniProt Entry Name: Q9UQT9_HUMAN

NCBI Description

HLA-DRB3 belongs to the HLA class II beta chain paralogues. This class II molecule is a heterodimer consisting of an alpha (DRA) and a beta (DRB) chain, both anchored in the membrane. It plays a central role in the immune system by presenting peptides derived from extracellular proteins. Class II molecules are expressed in antigen presenting cells (APC: B lymphocytes, dendritic cells, macrophages). The beta chain is approximately 26-28 kDa and its gene contains 6 exons. Exon one encodes the leader peptide, exons 2 and 3 encode the two extracellular domains, exon 4 encodes the transmembrane domain and exon 5 encodes the cytoplasmic tail. Within the DR molecule the beta chain contains all the polymorphisms specifying the peptide binding specificities. Typing for these polymorphisms is routinely done for bone marrow and kidney transplantation. DRB1 is expressed at a level five times higher than its paralogues DRB3, DRB4 and DRB5. The presence of DRB3 is linked with allelic variants of DRB1, otherwise it is omitted. There are 4 related pseudogenes: DRB2, DRB6, DRB7, DRB8 and DRB9. [provided by RefSeq, Jul 2008]

Uniprot Description

HLA-DRB3: Binds peptides derived from antigens that access the endocytic route of antigen presenting cells (APC) and presents them on the cell surface for recognition by the CD4 T-cells. The peptide binding cleft accommodates peptides of 10-30 residues. The peptides presented by MHC class II molecules are generated mostly by degradation of proteins that access the endocytic route, where they are processed by lysosomal proteases and other hydrolases. Exogenous antigens that have been endocytosed by the APC are thus readily available for presentation via MHC II molecules, and for this reason this antigen presentation pathway is usually referred to as exogenous. As membrane proteins on their way to degradation in lysosomes as part of their normal turn-over are also contained in the endosomal/lysosomal compartments, exogenous antigens must compete with those derived from endogenous components. Autophagy is also a source of endogenous peptides, autophagosomes constitutively fuse with MHC class II loading compartments. In addition to APCs, other cells of the gastrointestinal tract, such as epithelial cells, express MHC class II molecules and CD74 and act as APCs, which is an unusual trait of the GI tract. To produce a MHC class II molecule that presents an antigen, three MHC class II molecules (heterodimers of an alpha and a beta chain) associate with a CD74 trimer in the ER to form a heterononamer. Soon after the entry of this complex into the endosomal/lysosomal system where antigen processing occurs, CD74 undergoes a sequential degradation by various proteases, including CTSS and CTSL, leaving a small fragment termed CLIP (class-II-associated invariant chain peptide). The removal of CLIP is facilitated by HLA-DM via direct binding to the alpha-beta-CLIP complex so that CLIP is released. HLA-DM stabilizes MHC class II molecules until primary high affinity antigenic peptides are bound. The MHC II molecule bound to a peptide is then transported to the cell membrane surface. In B-cells, the interaction between HLA-DM and MHC class II molecules is regulated by HLA-DO. Primary dendritic cells (DCs) also to express HLA-DO. Lysosomal miroenvironment has been implicated in the regulation of antigen loading into MHC II molecules, increased acidification produces increased proteolysis and efficient peptide loading. Belongs to the MHC class II family.

Protein type: Membrane protein, integral

Chromosomal Location of Human Ortholog: 6p21.3

Cellular Component: Golgi membrane; membrane; lysosomal membrane; late endosome membrane; integral to plasma membrane; plasma membrane; trans-Golgi network membrane; external side of plasma membrane; MHC class II protein complex

Molecular Function: MHC class II receptor activity; protein binding; peptide antigen binding

Biological Process: T-helper 1 type immune response; detection of bacterium; cytokine and chemokine mediated signaling pathway; antigen processing and presentation of exogenous peptide antigen via MHC class II; signal transduction; immunoglobulin production during immune response; T cell receptor signaling pathway; humoral immune response mediated by circulating immunoglobulin; negative regulation of T cell proliferation; inflammatory response to antigenic stimulus; regulation of interleukin-4 production; negative regulation of interferon-gamma production; T cell costimulation; immune response; protein tetramerization