"Staining Solutions"

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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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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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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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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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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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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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This antibody reacts with an extracellular domain (close to transmembrane) of NCAM (Neural cell adhesion molecule), also called CD56. Three isoforms of NCAM/CD56 are produced by differential splicing of the RNA transcript from a single gene. The 135 kDa isoform is the basic molecule, which is glycosylated or sialylated to produce the mature species. NCAM antibody recognises two proteins of the neural cell adhesion molecule, the basic molecule expressed on most neuroectodermally derived tissues and neoplasms (e.g. retinoblastoma, medulloblastomas, astrocytomas, neuroblastomas, and small cell carcinomas). It is also expressed on some mesodermally derived tumors (rhabdomyosarcoma). NCAM antibody plays an important role in the diagnosis of nodal and nasal NK/T-cell lymphomas.

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Expression of TMEM16A (DOG1) protein is elevated in the gastrointestinal stromal tumors (GIST’s), c-kit signaling-driven mesenchymal tumors of the GI tract. It is rarely expressed in other soft tissue tumors, which, due to appearance, may be difficult to diagnose. Immunoreactivity for TMEM16A has been reported in 97,8 percent of scorable GIST’s, including all c-kit negative GIST’s. Overexpression has been suggested to aid in the identification of GISTs, including Platelet-Derived Growth Factor Receptor Alpha (PDGFR alpha) mutants that fail to express c-kit antigen. The overall sensitivity of TMEM16A and c-kit in GIST’s is nearly identical: 94,4% vs. 94,7%.

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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.

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IgM is the first antibody generated in an immune response to an antigen. It is generally a pentamer with each of the five immunoglobulins linked together with disulfide bonds. In its pentamer form, it has a molecular mass of 970 kDa and 10 antigen binding sites (due to the large size of most antigens, not all binding sites can be filled simultaneously.  IgM antibodies account for approximately 5%-10% of all the antibody in the body.

This mAb is specific for the human IgM heavy chain. It does not cross-react with other immunoglobulin heavy chains, T-cells, monocytes, granulocytes, or erythrocytes. IgM antibody is useful in the identification of certain cancer types. Some tumors express a single heavy chain class. Demonstration of clonality in lymphoid infiltrates indicates that the infiltrate is clonal and therefore malignant.

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It recognises a 66 kDa protein, which is identified as Tripartite motif-containing protein 29 (TRIM29). It interacts with the intermediate filament protein vimentin, a substrate for the PKC family of protein kinases, and with hPKCI-1, an inhibitor of the PKCs. TRIM29 protein contains both zinc finger and leucine zipper motifs, suggesting that the it may form homodimers and possibly associate with DNA. High expression of TRIM29 has been reported in gastric cancer and pancreatic cancer, and correlates with enhanced tumor growth and lymph node metastasis. TRIM29 is also able to distinguish lung squamous cell carcinoma from lung adenocarcinoma with ~90% positive accuracy, when used in a panel with TTF-1, p63, CK5/6, and Napsin-A antibodies.

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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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Actin participates in many important cellular processes, including muscle contraction, cell motility, cell division and cytokinesis, vesicle and organelle movement, cell signaling, and the establishment and maintenance of cell junctions and cell shape. An actin protein's mass is roughly 42- kDa and it is the monomeric subunit of two types of filaments in cells: microfilaments, one of the three major components of the cytoskeleton, and thin filaments, part of the contractile apparatus in muscle cells. It can be present as either a free monomer called G-actin (globular) or as part of a linear polymer microfilament called F-actin (filamentous), both of which are essential for such important cellular functions as the mobility and contraction of cells during cell division. [Wiki]

This antibody recognises actin of skeletal, cardiac, and smooth muscle cells.

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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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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.