Technical Data
H6098-34
HLA Class 1 Antigen A25,A32
Description:
The human leukocyte antigen system (HLA) is the name of the major histocompatibility complex (MHC) in humans. The superlocus contains a large number of genes related to immune system function in humans. This group of genes resides on chromosome 6, and encodes cell-surface antigen-presenting proteins and many other genes. The HLA genes are the human versions of the MHC genes that are found in most vertebrates (and thus are the most studied of the MHC genes). The proteins encoded by certain genes are also known as antigens, as a result of their historic discovery as factors in organ transplantations. The major HLA antigens are essential elements for immune function. Different classes have different functions: HLA class I antigens (A, B & C) present peptides from inside the cell (including viral peptides if present). These peptides are produced from digested proteins that are broken down in the proteasomes. The peptides are generally small polymers, about 9 amino acids in length. Foreign antigens attract killer T-cells (also called CD8 positive- or cytotoxic T-cells) that destroy cells.

HLA class II antigens (DP,DM, DOA,DOB,DQ, & DR) present antigens from outside of the cell to T-lymphocytes. These particular antigens stimulate T-helper cells to multiply, and these T-helper cells then stimulate antibody-producing B-cells to produce Antibodies to that specific antigen. Self-antigens are suppressed by suppressor T-cells.

HLA class III antigens encode components of the complement system.HLA have other roles. They are important in disease defense. They may be the cause of organ transplant rejections. They may protect against or fail to protect (if down regulated by an infection) cancers. They may mediate autoimmune disease (examples: type I diabetes, coeliac disease). Also, in reproduction, HLA may be related to the individual smell of people and may be involved in mate selection. Aside from the genes encoding the 6 major antigens, there are a large number of other genes, many involved in immune function, located on the HLA complex. Diversity of HLA in human population is one aspect of disease defense, and, as a result, the chance of two unrelated individuals having identical HLA molecules on all loci is very low. Historically, HLA genes were identified as a result of the ability to successfully transplant organs between HLA similar individuals.

Applications:
This antibody has not been tested in methodologies other than Microcytotoxicity Test. Potential applications include Flow Cytometry, Cell Typing, Tissue Staining and Chimerism Studies.

Recommended Dilutions:
Cytotoxicity: Add 0.9ml of 1% BSA in PBS to 100ul of10X working dilution.
Optimal dilutions to be determined by the researcher.

Cytotoxicity:
Expected Results:
Cell death will occur in any test well where 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.
Limitations:
Cell isolation difficulties, contamination of the lymphocyte preparation with red blood cells, monocytes, platelets or granulocytes, cell concentrations outside acceptable levels, bacterial contamination and/or change in pH of antisera may cause erroneous results.

Source:
Ascites fluid

Storage and Stability:
Lyophilized powder may be stored at -20C. Stable for 12 months at -20C. Reconstitute with sterile ddH2O. Aliquot to avoid repeated freezing and thawing. Store at -20C. Reconstituted product is stable for 12 months at -20C. 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.
TypeIsotypeCloneGrade
MabIgM4i99Ascites
SizeStorageShippingSourceHost
100ul-20CBlue IceHumanMouse
Concentration:
Not Determined
Immunogen:
Purity:
Ascites
Form
Supplied as a lyophilized powder from PBS, 1% BSA. Reconstitute with 100ul sterile dH2O to make a 10X stock solution.
Specificity:
Recognizes HLA Class 1 Antigen-A25,A32. Specificity was determined at a 1:10 dilution by the microcytotoxicity test under standard NIH conditions.
Intended for research use only. Not for use in human, therapeutic, or diagnostic applications.
1. NCCLS Tentative Standard. "Leukocyte Differential Counting" Publication Number H20-T, NCCLS, Vol. 4, No. 11 (1984). 2. Pei, R., et al., Amonospecific HLA-27 fluorescein isothiocyanate conjugated monoclonal antibody for rapid, simple and accurate HLA-B27 typing. Tissue Antigen 41: 200-203 (1993). 3. Pei, R., Woo, G. and Lee, J.H. "Detection of blood chimerism at a frequency of one per thousand by flow cytometry. Visuals of the clinical histocompatibility workshop", Paul I. Terasaki, Ed., pp.73-74 (1995). 4. Pei, R., Chen, T., Orpilla, J. and Lee, J.H. "A simultaneous negative and positive selection method that can detect chimerism at a frequency of 1 per 10,000 by flow cytometry", Tissue Antigens 50:197-201 (1997).