"Staining Solutions"

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CD8 is a cell surface receptor expressed either as a heterodimer with the beta chain or as a homodimer. A majority of thymocytes and a subpopulation of mature Tcells and NK cells express CD8a. It binds to MHC class 1 and through its association with protein tyrosine kinase p56lck plays a role in Tcell development and activation of mature Tcells. For mature Tcells, CD8 and 4 are mutually exclusive, so antibody to CD8 is generally used in conjunction with antibody to CD4. CD8a antibody is a useful marker for distinguishing helper/inducer T-lymphocytes, and most peripheral Tcell lymphomas are CD4+/CD8-. Anaplastic large cell lymphoma is usually CD4+/CD8-, and in T-lymphoblastic lymphoma/leukemia, they are often co-expressed. It is also found in littoral cell angioma of the spleen.

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Recognises a 60 kDa protein, identified as the heat shock protein 60 (Hsp60). Its epitope is localized between amino acids 383-447 of human Hsp60. A wide variety of environmental and pathophysiological stressful conditions trigger the synthesis of a family of proteins known as heat shock proteins (hsps), more appropriately called as stress response proteins (srps). Hsp60 is a potential antigen in a number of autoimmune diseases. In human arthritis and in experimentally induced arthritis in animals, disease development coincides with the development of immune reactivity directed against not only bacterial Hsp60, but also against its mammalian homolog.

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Recognises a carbohydrate determinant of Gal 1-3(Fuc 1-4) GlcNAc which is blood group antigen Lewis A. It is present primarily on epithelial cells such as colon and kidneys. In the tumors and dedifferentiated tissues, decrease of Lewis A antigen was observed. Lewis A (type 1 chain) is expressed in colonic epithelial cells and may be useful for detection of gastrointestinal tumors, pancreatic cancer, and colorectal tumors. Blood group related antigens represent a group of carbohydrate determinants carried on both glycolipids and glycoproteins. They are usually mucin-type, and are detected on erythrocytes, certain epithelial cells, and in secretions of certain individuals. Sixteen genetically and biosynthetically distinct but inter-related specificities belong to this group of antigens, including A, B, H, Lewis A, Lewis B, Lewis X, Lewis Y, and precursor type 1 chain antigens.

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Recognises a protein of 210-220 kDa, which is identified as the Complement receptor 1 or CD35. This mAb is specific for a site in CR1 that is distal from the C3b-binding site, so that it is unable to block CR1/CD35 activity. It is highly specific and shows no cross-reaction with CR2/CD21. The primary function of CR1 is to serve as the cellular receptor for C3b and C4b, the most important components of the complement system leading to clearance of foreign macromolecules. The Knops blood group system is a system of antigens located on this protein. Follicular dendritic cells (FDC) are restricted to the B-cell regions of secondary lymphoid follicles. They are CD21+/CD35+/CD1a-. This mAb labels follicular dendritic cells and follicular dendritic cell sarcoma.

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The tetraspanins are integral membrane proteins expressed on cell surface and granular membranes of hematopoietic cells and are components of multi-molecular complexes with specific integrins. The tetraspanin CD63 is a lysosomal membrane glycoprotein that translocates to the plasma membrane after platelet activation. CD63 is expressed on activated platelets, monocytes and macrophages, and is weakly expressed on granulocytes, T cell and B cells. It is located on the basophilic granule membranes and on the plasma membranes of lymphocytes and granulocytes. CD63 is a member of the TM4 superfamily of leukocyte glycoproteins that includes CD9, CD37 and CD53, which contain four transmembrane regions. CD63 may play a role in phagocytic and intracellular lysosome-phagosome fusion events. CD63 deficiency is associated with Hermansky-Pudlak syndrome and is strongly expressed during the early stages of melanoma progression.

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This mAb recognises a 21 kDa protein, identified as the p21WAF1 tumor suppressor protein. This mAb is highly specific to p21 and shows no cross-reaction with other closely related mitotic inhibitors. p21WAF1 is a specific inhibitor of cdk s and a tumor suppressor involved in the pathogenesis of a variety of malignancies. The expression of this gene acts as an inhibitor of the cell cycle during G1 phase and is tightly controlled by the tumor suppressor protein p53. Its expression is induced by the wild type, but not mutant, p53 suppressor protein. Normal cells generally display a rather intense nuclear p21 expression. Loss of p21 expression has been reported in many carcinomas (gastric carcinoma, non-small cell lung carcinoma, thyroid carcinoma).

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This mAb recognises a 21 kDa protein, identified as the p21WAF1 tumor suppressor protein. This mAb is highly specific to p21 and shows no cross-reaction with other closely related mitotic inhibitors. p21WAF1 is a specific inhibitor of cdk s and a tumor suppressor involved in the pathogenesis of a variety of malignancies. The expression of this gene acts as an inhibitor of the cell cycle during G1 phase and is tightly controlled by the tumor suppressor protein p53. Its expression is induced by the wild type, but not mutant, p53 suppressor protein. Normal cells generally display a rather intense nuclear p21 expression. Loss of p21 expression has been reported in many carcinomas (gastric carcinoma, non-small cell lung carcinoma, thyroid carcinoma).

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Basigin (BSG) also known as extracellular matrix metalloproteinase inducer (EMMPRIN) or cluster of differentiation 147 (CD147) is a protein that in humans is encoded by the BSG gene. This protein is a determinant for the Ok blood group system. Basigin has been shown to be an essential receptor on red blood cells for the human malaria parasite, Plasmodium falciparum. It has a variety of functions. In addition to its metalloproteinase-inducing ability, basigin also regulates several distinct functions, such as spermatogenesis, expression of the monocarboxylate transporter and the responsiveness of lymphocytes. CD147 is a type I integral membrane receptor that has many ligands, including the cyclophilin (CyP) proteins Cyp-A and CyP-B and certain integrins. It is expressed by many cell types, including epithelial cells, endothelial cells and leukocytes. [Wiki].

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Recognises a glycoprotein of ~200 kDa, identified as carbonic anhydrase IX (CAIX/CA9/gp200). Its epitope resides in the carbohydrate domain of gp200. It shows no significant cross-reactivity with other carbohydrate determinants, such as the Lewis blood group antigens, epithelial membrane antigen, HMFG, and AB blood group antigens. In normal kidney, gp200 is localized along the brush border of the pars convoluta and pars recta segments of the proximal tubule, as well as focally along the luminal surface of Bowman's capsule adjoining the outgoing proximal tubule. Reportedly, gp200 is expressed by 93% of primary and 84% of metastatic renal cell carcinomas. This mAb may be useful in the investigations of carcinomas of proximal nephrogenic differentiation especially those showing tubular differentiation.

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Estrogen receptor (ER) is an important regulator of growth and differentiation in the mammary gland. It is found in two forms, alpha and beta. Both forms are widely expressed, however, alpha is the predominant form in endometrium and breast cancer cells, and beta is more highly expressed in brain, kidney, heart, lung and bone. Activated Estrogen receptor dimerizes and forms both homo- and heterodimers. ER contains five domains: A-F. The A and B domain are able to weakly activate ER in the absence of ligand. The C domain binds DNA and the E domain contains the ligand binding site. Presence of ER in breast tumors indicates an increased likelihood of response to anti-estrogen (e.g. tamoxifen) therapy. This antibody is specific to estrogen receptor alpha and shows minimal cross-reaction with other members of the family.

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CD45R, also designated CD45 and PTPRC, has been identified as a transmembrane glycoprotein, broadly expressed among hematopoietic cells. Multiple isoforms of CD45R are distributed throughout the immune system according to cell type. These isoforms arise because of alternative splicing of exons 4, 5, and 6. The corresponding protein domains are characterised by the binding of monoclonal antibodies specific for CD45RA (exon 4), CD45RB (exon 5), CD45RC (exon 6) and CD45RO (exons 4 to 6 spliced out). The variation in these isoforms is localized to the extracellular domain of CD45R, while the intracellular domain is conserved. CD45RB is expressed on mature B-lymphocytes and the majority of lymphomas and leukemias of B-cell origin. This antigen is also expressed on the surface of cells of myeloid lineage.

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Smooth muscle myosin heavy chain (SM-MHC) is a cytoplasmic structural protein, which is a major component of the contractile apparatus in smooth muscle cells. Expression of smooth muscle myosin is developmentally regulated, appearing early in smooth muscle development, and is specific for smooth muscle development. Two isoforms of smooth muscle myosin heavy chain have been identified, designated MHC-1 and MHC-2. The antibody may be useful for the study of breast tumors as the presence of an intact layer of myoepithelial cells is an important feature, which may distinguish benign breast lesions and carcinoma in situ from invasive tumors.

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The c-Myc protein is a transcription factor, which is encoded by the c-Myc gene on human chromosome 8q24. c-Myc is commonly activated in a variety of tumor cells and plays an important role in cellular proliferation, differentiation, apoptosis and cell cycle progression. The phosphorylation of c-Myc has been investigated and previous studies have suggested a functional association between phosphorylation at Thr58/Ser62 by glycogen synthase kinase 3, cyclin dependent kinase, ERK2 and C-Jun N terminal Kinase (JNK) in cell proliferation and cell cycle regulation. Studies also have shown that c-Myc is essential for tumor cell development in vasculogenesis and angiogenesis that distribute blood throughout the cells, and which brought extensive attention in the development of new therapeutic approach for cancer treatment.

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Cytokeratin-7 (CK7) is a member of the keratin gene family. The type II cytokeratins consist of basic or neutral proteins which are arranged in pairs of heterotypic keratin chains coexpressed during differentiation of simple and stratified epithelial tissues. CK7 is specifically expressed in the simple epithelia lining the cavities of the internal organs and in the gland ducts and blood vessels. It is found in simple glandular epithelia, and in transitional epithelium. Epithelial cells of the lung and breast both contain CK7, but some other glandular epithelia, such as those of the colon and prostate, do not. Because the protein is found in both healthy and neoplastic cells, antibodies to CK7 can be used in immunohistochemistry to distinguish ovarian and transitional cell carcinomas from colonic and prostate cancers, respectively. It is commonly used together with CK20 when making such diagnoses. [Wiki].

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This mAb is specific to SUMO-1 and shows no cross-reaction with either SUMO2 or SUMO3. The small ubiquitin-related modifier (SUMO) proteins, which include SUMO1, SUMO2 and SUMO3, belong to the ubiquitin-like protein family. Like ubiquitin, the SUMO proteins are synthesised is precursor proteins that undergo processing before conjugation to target proteins. Also, both utilize the E1, E2, and E3 cascade enzymes for conjugation. However, SUMO and ubiquitin differ with respect to targeting. Ubiquitination predominantly targets proteins for degradation, whereas sumoylation targets proteins to a variety of cellular processing, including nuclear transport, transcriptional regulation, apoptosis and protein stability. The unconjugated SUMO1 protein localizes to the nuclear membrane.

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Cyclins, regulatory subunits which associate with kinases, control many of the important steps in cell cycle progression. The Cdc2 kinase (p34Cdc2) exhibits kinase activity in vitro and exists in a complex with both cyclin B and a protein homologous to p13SUC1. Cdc2 kinase is the active subunit of the M phase promoting factor (MPF) and the M phase-specific Histone H1 kinase. The p34Cdc2/cyclin B complex is required for the G2 to M transition. An additional cell cycle-dependent protein kinase, termed p55cdc, exhibits a high degree of homology with the S. cerevisiae proteins Cdc20 and Cdc4. The p55cdc transcript is readily detectable in a variety of cultured cell lines in growth phase, but disappears when cell growth is chemically arrested.