"Staining Solutions"

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Proliferating cell nuclear antigen is an auxiliary protein of DNA polymerase delta and is involved in the control of eukaryotic DNA replication by increasing the polymerase's processibility during elongation of the leading strand. Induces a robust stimulatory effect on the 3'-5' exonuclease and 3'-phosphodiesterase, but not apurinic-apyrimidinic (AP) endonuclease, APEX2 activities. Plays a key role in DNA damage response (DDR) by being conveniently positioned at the replication fork to coordinate DNA replication with DNA repair and damage tolerance pathways. Acts as a loading platform to recruit DDR proteins that allow completion of DNA replication after damage and promote postreplication repair: Monoubiquitinated PCNA leads to recruitment of translesion (TLS) polymerases, while 'Lys-63'-linked polyubiquitination of PCNA is involved in error-free pathway and employs recombination mechanisms to synthesize across the lesion. [UniProt].

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Recognises a protein of 42-46 kDa, identified as MAGE-1. This mAb does not cross-react with other members of MAGE-family. Human malignant neoplasms carry rejection antigens that are recognized by the patients' autologous, tumor directed and specific, cytolytic, CD8+ T lymphocyte clones (CTL). The MAGE family of genes codes an important group of antigens. It was identified that melanomas and primary glial brain tumors express common melanoma associated antigens (MAAs). Because MAGE-1 is expressed on a significant proportion of human neoplasms of various histological types (melanoma, brain tumors of glial origin, neuroblastoma, non-small cell lung cancer, breast, gastric, colorectal, ovarian, renal cell carcinomas) and not on normal tissues, the encoded antigen may serve as a marker of early detection and target for cancer immunotherapy.

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CD45RA is an isoform of the human leukocyte common antigen (CD45). Human CD45 contains three exons which encode peptide segments designated A, B and C, respectively. The differential splicing of the exons generates at least five isoforms, ABC, AB, BC, B and O. This antibody reacts with ABC and BC isoforms. CD45RA is expressed on 40-50% of peripheral CD4+ T-cells, 50% of peripheral CD8+ T-cells, B-cells, and leukemic B-cell lines. T-cells expressing CD45RA are naive or virgin T-cells. T-cells expressing CD45RO are memory T-cells. CD45RA and CD45RO define complementary, predominantly non-overlapping populations of resting peripheral T-cells. This MAb is useful in study on the subpopulation of CD4+ or CD8+ T-cells. It can especially be used to differentiate T-cell lymphomas (CD45RO +ve) from B cell lymphomas (CD45RA +ve).

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This antibody recognises an oncofetal antigen of 220 kDa, identified as a tumor-associated glycoprotein (TAG-72) with properties of a mucin. This antibody defines the mucin-carried sialylated-Tn epitope. TAG-72 is usually expressed by adenocarcinomas, but is negative in mesotheliomas. Studies have reported that this antibody has 80% sensitivity and 93% specificity for pulmonary adenocarcinoma. Therefore, TAG-72 is a useful marker to distinguish between mesothelioma and adenocarcinoma. However, false positive reactions can occur so results must be interpreted with the utmost caution. This antibody may be useful in the differentiation of non-small cell carcinomas from small cell carcinomas of the lung. The combined use of TAG-72 antibody with a GCDFP-15 antibody is valuable in the diagnosis of apocrine carcinoma.

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In humans, multiple isoforms of Cytokeratin 6 (6A-6F), encoded by several highly homologous genes, have distinct tissue expression patterns, and Cytokeratin 6A is the dominant form in epithelial tissue. The gene encoding human Cytokeratin 6A maps to chromosome 12q13, and mutations in this gene are linked to several inheritable hair and skin pathologies. Keratins 6 and 16 are expressed in keratinocytes, which are undergoing rapid turnover in the suprabasal region (also known as hyper-proliferation-related keratins). Keratin 6 is found in hair follicles, suprabasal cells of a variety of internal stratified epithelia, in epidermis, in both normal and hyper-proliferative situations. Epidermal injury results in activation of keratinocytes, which express CK6 and CK16. CK6 is strongly expressed in about 75% of head and neck squamous cell carcinomas. Expression of CK6 is particularly associated with differentiation.

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This antibody recognises an oncofetal antigen of 220 kDa, identified as a tumor-associated glycoprotein (TAG-72) with properties of a mucin. This antibody defines the mucin-carried sialylated-Tn epitope. TAG-72 is usually expressed by adenocarcinomas, but is negative in mesotheliomas. Studies have reported that this antibody has 80% sensitivity and 93% specificity for pulmonary adenocarcinoma. Therefore, TAG-72 is a useful marker to distinguish between mesothelioma and adenocarcinoma. However, false positive reactions can occur so results must be interpreted with the utmost caution. This antibody may be useful in the differentiation of non-small cell carcinomas from small cell carcinomas of the lung. The combined use of TAG-72 Ab with a GCDFP-15 Ab is valuable in the diagnosis of apocrine carcinoma.

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It recognises a protein of HMW, identified as mucin 3 glycoprotein (MUC3). This mAb shows no cross-reaction with human milk fat globule membranes, MUC1, or MUC2. The Mucins are a family of highly glycosylated, secreted proteins with a basic structure consisting of a variable number of tandem repeats (VNTRs) encoded by 60 base pairs (Mucin 1), 69 base pairs (Mucin 2) and 51 base pairs (Mucin 3). The number of repeats is highly polymorphic and varies among different alleles. Mucin 1 proteins are expressed as type I membrane proteins in addition to secreted forms. Mucin 1 is aberrantly expressed in epithelial tumors including breast carcinomas. Mucin 2 coats the epithelia of the intestines and airways and is associated with colonic tumors. Mucin 3 is a major component of various mucus gels and is broadly expressed in normal and tumor cells.

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Reacts with human CD59, a 20 kDa glycosyl phosphatidyl-inositol (GPI)-anchored cell surface protein. CD59 regulates complement-mediated cell lysis, and it is involved in lymphocyte signal transduction. This protein is a potent inhibitor of the complement membrane attack complex, whereby it binds complement C8 and/or C9 during the assembly of this complex, thereby inhibiting the incorporation of multiple copies of C9 into the complex, which is necessary for osmolytic pore formation. CD59 is widely distributed on cells in all tissues. It inhibits formation of MAC, thus protecting cells from complement-mediated lysis. The expression of CD59 on erythrocytes is important for their survival. Genetic defects in GPI-anchor attachment, that cause a reduction or loss of CD59 and CD55 on erythrocytes produce the symptoms of the disease paroxysmal hemoglobinuria (PNH). It is useful for study on GPI-anchored proteins, PNH and CD59 functions.

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Thyroglobulin is a 660 kDa dimeric pre-protein with mutiple glycosylation sites. It is produced by and processed within the thyroid gland to produce the hormone thyroxine and triiodothyronine. Prior to forming dimers, thyroglobulin monomers undergo conformational maturation in the endoplasmic reticulation. The vast majority of follicular carcinomas of the thyroid will give positive immunoreactivity for anti-thyroglobulin even though sometimes only focally. Poorly differentiated carcinomas of the thyroid are frequently anti-thyroglobulin negative. Adenocarcinomas of other-than-thyroid origin do not react with this antibody. This antibody is useful in identification of thyroid carcinoma of the papillary and follicular types. Presence of thyroglobulin in metastatic lesions establishes the thyroid origin of tumor. Anti-thyroglobulin, combined with anti-calcitonin, can identify medullary carcinomas of the thyroid. Furthermore, anti-thyroglobulin, combined with anti-TTF1, can be a reliable marker to differentiate between primary thyroid and lung neoplasms.

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This mAb recognises a protein of 130 kDa, identified as Ksp-cadherin. Cadherins form a superfamily of related glycoproteins that mediate calcium-dependent cell adhesion and transmit signals from the extracellular matrix to the cytoplasm. Cadherins have been implicated in embryogenesis, tissue morphogenesis, tissue structure maintenance, cell polarization, neoplastic invasiveness and metastasis, and membrane transport. It is suggested that Ksp-cadherin is a marker for terminal differentiation of the basolateral membranes of renal tubular epithelial cells. Within the kidney, Ksp-Cadherin is found exclusively in the basolateral membrane of renal tubular epithelial cells and collecting duct cells, and not in glomeruli, renal interstitial cells, or blood vessels. Ksp-Cadherin has been suggested to distinguish Chromophobe Renal-Cell Carcinoma from Oncocytoma.

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PECAM-1, or platelet endothelial cell adhesion molecule-1, also called CD31, is made up of six extracellular immunoglobulin-like domains, a single transmembrane domain, and a cytoplasmic domain containing serine and tyrosine residues that when phosphorylated, activate the molecules signaling function. It is expressed on platelets, monocytes, neutrophils, bone marrow macrophage and dendritic cells precursors, and at the intracellular junction of endothelial cells. PECAM-1, on the endothelial lining of blood vessels, helps maintain the vascular barrier. In response to inflammatory mediators, the endothelial pull away from each other allowing fluid, inflammatory cells and proteins into the extravascular space. The inflammatory response fades as adhesion of neighboring cells increase. PECAM-1 also plays an important role in angiogenesis, promoting endothelial cell motility, junctional development and capillary morphogenesis.

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CD45RA is an isoform of the human leukocyte common antigen (CD45). Human CD45 contains three exons which encode peptide segments designated A, B and C, respectively. The differential splicing of the exons generates at least five isoforms, ABC, AB, BC, B and O. This antibody reacts with ABC and BC isoforms. CD45RA is expressed on 40-50% of peripheral CD4+ T-cells, 50% of peripheral CD8+ T-cells, B-cells, and leukemic B-cell lines. T-cells expressing CD45RA are naive or virgin T-cells. T-cells expressing CD45RO are memory T-cells. CD45RA and CD45RO define complementary, predominantly non-overlapping populations of resting peripheral T-cells. This MAb is useful in study on the subpopulation of CD4+ or CD8+ T-cells. It can especially be used to differentiate T-cell lymphomas (CD45RO +ve) from B cell lymphomas (CD45RA +ve).

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This mAb recognises a protein of 165 kDa, identified as carbamoyl phosphate synthetase 1 (CPS1). This mitochondrial enzyme catalyses synthesis of carbamoyl phosphate from ammonia and bicarbonate. This reaction is the first committed step of the urea cycle, which is important in the removal of excess urea from cells. Deficiency of CPS1 is an autosomal recessive disorder that causes hyperammonemia. CPS1 is a hepatocyte specific protein that localizes to the mitochondria of hepatocytes. It is a sensitive marker for distinguishing hepatocellular carcinomas (HCC) from other metastatic carcinomas as well as cholangio-carcinomas. HCC s occur primarily in the stomach, but they are also found in many other organs. CPS1 may also be a useful marker for intestinal metaplasia. Reportedly, strong expression of CPS1 correlates with smaller tumor size and longer patient survival. Occasionally, CPS1 is also found in gastric carcinomas as well as in a few other non-hepatic tumors.

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Recognises a protein of 75 kDa, identified as mu heavy chain of human immunoglobulins. It does not cross-react with alpha (IgA), gamma (IgG), epsilon (IgE), or delta (IgD), heavy chains, T-cells, monocytes, granulocytes, or erythrocytes. Monomeric IgM is expressed as a membrane bound antibody on the surface of B cells and as a pentamer when secreted by plasma cells. IgM antibody is prominent in early immune responses to most antigens. Aberrant levels are associated with immune deficiency states, hereditary deficiencies, myeloma, Waldenstrom's macroglobulinemia, chronic infection and hepatocellular disease. This mAb is useful in the identification of leukemias, plasmacytomas, and certain non-Hodgkins lymphomas. The most common feature of these malignancies is the restricted expression of a single heavy chain class. Demonstration of clonality in lymphoid infiltrates indicates that the infiltrate is clonal and therefore malignant.

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This antibody recognises a glycoprotein of ~200 kDa, identified as renal cell carcinoma protein, carbonic anhydrase IX/CAIX, or gp200. Its epitope resides in the carbohydrate domain of the protein. The CAIX antibody 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, CAIX 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, it is expressed by 93% of primary and 84% of metastatic renal cell carcinomas. This CAIX antibody may be useful in the investigations of carcinomas of proximal nephrogenic differentiation, especially those showing tubular differentiation.

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This antibody reacts with a protein of 22 kDa, identified as beta subunit of HCG. It does not cross react with the alpha subunit. HCG is a glycoprotein which is secreted in large quantities by normal trophoblasts. It is present only in trace amounts in non-pregnant urine and sera but rises sharply during pregnancy. HCG is composed of two non-identical, non-covalently linked polypeptide chains designated as the alpha and beta subunits. The alpha subunit is identical to that of thyroid stimulating hormone (TSH), follicle stimulating hormone (FSH), and luteinizing hormone (LH). HCG-beta antibody detects cells and tumors of trophoblastic origin such as choriocarcinoma. Large cell carcinoma and adenocarcinoma of the lung demonstrate antibody positivity in 90% and 60% of cases respectively; 20% of lung squamous cell carcinomas are positive. HCG expression by non-trophoblastic tumors may indicate aggressive behavior.