"Staining Solutions"

Supplier: ProSci Inc.

This antibody recognises a protein of 205 kDa-220 kDa, identified as CD45RA, an isoform of the human leukocyte common antigen (CD45). Human CD45 contains three exons which encode peptide segments designated A, B and C. 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+ Tcells, 50% of peripheral CD8+ T-cells, B-cells, and leukemic B-cell lines. T-cells expressing CD45RA are naive or virgin Tcells. T-cells expressing CD45RO are memory Tcells. CD45RA and CD45RO define complementary, predominantly non-overlapping populations of resting peripheral Tcells. This CD45RA antibody is useful in the study of the subpopulation of CD4+ or CD8+ T-cells. It is especially useful in differentiating T-cell lymphomas (CD45RO +ve) from B cell lymphomas (CD45RA +ve).

Supplier: ProSci Inc.

The Cell-division cycle protein 20 is an essential regulator of cell division that is encoded by the CDC20 gene in humans. To the best of current knowledge its most important function is to activate the anaphase promoting complex (APC/C), a large 11-13 subunit complex that initiates chromatid separation and entrance into anaphase. The APC/C-Cdc20 protein complex has two main downstream targets. Firstly, it targets securin for destruction, enabling the eventual destruction of cohesin and thus sister chromatid separation. It also targets S and M-phase (S/M) cyclins for destruction, which inactivates S/M cyclin-dependent kinases (Cdks) and allows the cell to exit from mitosis. A closely related protein, Cdc20homologue-1 (Cdh1) plays a complementary role in the cell cycle.

Cdc20 appears to act as a regulatory protein interacting with many other proteins at multiple points in the cell cycle. It is required for two microtubule-dependent processes: nuclear movement prior to anaphase, and chromosome separation. [Wiki].

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Mucin-1 is a large cell surface mucin glycoprotein expressed by most glandular and ductal epithelial cells and some hematopoietic cell lineages. It is expressed on most secretory epithelium, including mammary gland and some hematopoietic cells. It is expressed abundantly in lactating mammary glands and over expressed in >90% breast carcinomas and metastases. The transgenic protein has been shown to associate with all four c-erbB receptors and localize with c-erbB1 (EGFR) in lactating glands. The gene contains seven exons and produces several different alternatively spliced variants. The major expressed form of the protein uses all seven exons and is a type 1 transmembrane protein with a large extracellular tandem repeat domain. The tandem repeat domain is highly O glycosylated and alterations in glycosylation have been shown in epithelial cancer cells. Mucin-1 antibody is useful as a pan-epithelial marker for detecting early metastatic loci of carcinoma in bone marrow or liver.

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Recognises a sialoglycoprotein of 27 to 32 kDa, identified as CD99, or MIC2 gene product, or E2 antigen. MIC2 gene is located in the pseudo-autosomal region of the human X and Y chromosome. MIC2 gene encodes two distinct proteins, which are produced by alternative splicing of the CD99 gene transcript and are identified as bands of 30 and 32 kDa (p30/32). Although its function is not fully understood, CD99 is implicated in various cellular processes including homotypic aggregation of T cells, upregulation of T cell receptor and MHS molecules, apoptosis of immature thymocytes and leukocyte diapedesis. CD99 is expressed on the cell membrane of some lymphocytes, cortical thymocytes, and granulosa cells of the ovary. Most pancreatic islet cells, Sertoli cells of the testis, and some endothelial cells express this antigen. Mature granulocytes express very little or no CD99. MIC2 is strongly expressed on Ewing s sarcoma cells and primitive peripheral neuroectodermal tumors.

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Tyrosine-protein kinase ZAP70 is a tyrosine kinase that plays an essential role in regulation of the adaptive immune response. Regulates motility, adhesion and cytokine expression of mature T-cells, as well as thymocyte development. Contributes also to the development and activation of primary B-lymphocytes. When antigen presenting cells (APC) activate T-cell receptor (TCR), a serie of phosphorylations lead to the recruitment of ZAP70 to the doubly phosphorylated TCR component CD247/CD3Z through ITAM motif at the plasma membrane. This recruitment serves to localization to the stimulated TCR and to relieve its autoinhibited conformation. Release of ZAP70 active conformation is further stabilized by phosphorylation mediated by LCK. Subsequently, it phosphorylates at least 2 essential adapter proteins: LAT and LCP2. In turn, a large number of signaling molecules are recruited and ultimately lead to lymphokine production, T-cell proliferation and differentiation. Furthermore, ZAP70 controls cytoskeleton modifications, adhesion and mobility of T-lymphocytes, thus ensuring correct delivery of effectors to the APC. [UniProt].

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Cytokeratin 8 (CK8) belongs to the type II (or B or basic) subfamily of high molecular weight cytokeratins and exists in combination with cytokeratin 18 (CK18). Cytokeratin 8 is primarily found in the non-squamous epithelia and is present in majority of adenocarcinomas and ductal carcinomas. It is absent in squamous cell carcinomas. Hepatocellular carcinomas are defined by the use of antibody that recognises only cytokeratin 8 and 18. Cytokeratin 8 exists on several types of normal and neoplastic epithelia, including many ductal and glandular epithelia such as colon, stomach, small intestine, trachea, and esophagus as well as in transitional epithelium. Antibody to Cytokeratin 8 does not react with skeletal muscle or nerve cells. Epithelioid sarcoma, chordoma, and adamantinoma show strong positivity corresponding to that of simple epithelia (with antibodies against Cytokeratin 8, 18 and 19). Reportedly, Cytokeratin 8 antibody is useful for the differentiation of lobular (“ring-like, perinuclear”) from ductal (“peripheral-predominant”) carcinoma of the breast.

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Recognises a protein of 47-55 kDa, which is identified as FOXP3. Its precise epitope is not known, but it has been mapped to the N-terminal portion of the protein. The FOX family of transcription factors is a large group of proteins that share a common DNA binding domain termed a winged-helix or forkhead domain. During early development, FOXP1 and FOXP2 are expressed abundantly in the lung, with lower levels of expression in neural, intestinal and cardiovascular tissues, where they act as transcription repressors. FOXP1 is widely expressed in adult tissues, while neoplastic cells often exhibit a dramatic change in expression level or localization of FOXP1. Mutations in FOXP3 gene cause IPEX, a fatal, X-linked inherited disorder characterised by immune dysregulation. The FOXP3 protein is essential for normal immune homeostasis. Specifically, FOXP3 represses transcription through a DNA binding forkhead domain, thereby regulating T cell activation.

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Recognises a sialoglycoprotein of 27 to 32 kDa, identified as CD99, or MIC2 gene product, or E2 antigen. MIC2 gene is located in the pseudo-autosomal region of the human X and Y chromosome. MIC2 gene encodes two distinct proteins, which are produced by alternative splicing of the CD99 gene transcript and are identified as bands of 30 and 32 kDa (p30/32). Although its function is not fully understood, CD99 is implicated in various cellular processes including homotypic aggregation of T cells, upregulation of T cell receptor and MHS molecules, apoptosis of immature thymocytes and leukocyte diapedesis. CD99 is expressed on the cell membrane of some lymphocytes, cortical thymocytes, and granulosa cells of the ovary. Most pancreatic islet cells, Sertoli cells of the testis, and some endothelial cells express this antigen. Mature granulocytes express very little or no CD99. It is strongly expressed on Ewing s sarcoma cells and primitive peripheral neuroectodermal tumors.

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Thyroglobulin is a 660 kDa dimeric pre-protein with mutiple glycosylation sites, detected at ~300 kDa in western blot. 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 thyroglobulin antibody even though sometimes only focally. Poorly differentiated carcinomas of the thyroid are frequently thyroglobulin antibody 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. Thyroglobulin antibody, combined with calcitonin antibody, can identify medullary carcinomas of the thyroid. Furthermore, thyroglobulin antibody, combined with TTF1 antibody, can be a reliable marker to differentiate between primary thyroid and lung neoplasms.

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At least five CD1 genes (CD1a, b, c, d, and e) are identified. CD1 proteins have been demonstrated to restrict Tcell response to non-peptide lipid and glycolipid antigens and play a role in non-classical antigen presentation. CD1a is a non-polymorphic MHC Class 1 related cell surface glycoprotein, expressed in association with Beta-2 microglobulin. CD1a antibody labels Langerhans cell histiocytosis (Histiocytosis X), extranodal histiocytic sarcoma, a subset of T-lymphoblastic lymphoma/leukemia, and interdigitating dendritic cell sarcoma of the lymph node. When combined with antibody against TTF-1 and CD5, CD1a antibody is useful in distinguishing between pulmonary and thymic neoplasms since it is consistently expressed in thymic lymphocytes in both typical and atypical thymomas, but only focally in 1/6 of thymic carcinomas and not in lymphocytes in pulmonary neoplasms. CD1a antibody is reported to be a new marker for perivascular epithelial cell tumor (PEComa).

Supplier: ProSci Inc.

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 granulocyte-colony stimulating factor (G-CSF) in the cytoplasm of mature granulocytes. It shows no reactivity with any other cell types. Markers of myeloid cells are useful in the identification of different levels of cellular differentiation. It reacts with early precursor and mature forms of myeloid cells. It is useful for the detection of myeloid leukemias and granulocytic sarcomas. It can be used as a marker of granulocytes in normal tissues or inflammatory processes.G-CSF is a pleiotropic cytokine that influences differentiation, proliferation and activation of the neutrophilic granulocyte lineage. The human G-CSF cDNA encodes a 207 amino acid precursor containing a 29 amino acid signal peptide that is proteolytically cleaved to form a 178 amino acid residue mature protein. Two G-CSFs, which are identical except for a three amino acid deletion in the amino-terminus of one form of the protein have been isolated from human cells. Murine and human G-CSF s share 73% sequence identity at the amino acid level.

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At least five CD1 genes (CD1a, b, c, d, and e) are identified. CD1 proteins have been demonstrated to restrict T cell response to non-peptide lipid and glycolipid antigens and play a role in non-classical antigen presentation. CD1a is a non-polymorphic MHC Class 1 related cell surface glycoprotein, expressed in association with Beta-2 microglobulin. CD1a antibody labels Langerhans cell histiocytosis (Histiocytosis X), extranodal histiocytic sarcoma, a subset of T-lymphoblastic lymphoma/leukemia, and interdigitating dendritic cell sarcoma of the lymph node. When combined with antibody against TTF-1 and CD5, CD1a antibody is useful in distinguishing between pulmonary and thymic neoplasms since CD1a is consistently expressed in thymic lymphocytes in both typical and atypical thymomas, but only focally in 1/6 of thymic carcinomas and not in lymphocytes in pulmonary neoplasms. antibody to CD1a is reported to be a new marker for perivascular epithelial cell tumor (PEComa).

Supplier: Biosensis

Monoclonal antibody MC192 against the rat low affinity nerve growth factor receptor (p75NTR) is derived from the fusion of Sp2/0-Ag 14 myeloma cells with mouse immune splenocytes. MC192 monoclonal antibody was originally generated by Chandlers et al. p75NTR was originally discovered as a low affinity nerve growth factor receptor. Later it was found that it was the receptor for all neurotrophins. It mediates signals of neurotrophins for neuronal survival, apoptosis, neurite outgrowth and synaptic plasticity. Recently, it has been revealed that p75NTR is not only acts as the receptor for neurotrophins but also the receptor for many other pathological ligands such as prions, rabies virus and amyloid beta. p75NTR also acts as a co-receptor for NOGO which mediates inhibitory signals of myelin associated protein. p75NTR is highly expressed in a number of non-neuronal and neuronal cells including motor neurons during development and also in damaged neurons. MC192 recognizes the extracellular domain of the neurotrophin receptor p75NTR in rat. MC192 antibody may be used for immunocytochemical localisation of rat cells expressing p75NTR, ELISA and western blot. This antibody has also been used for the construction of the MC192-saporin immunotoxin for specific elimination of neuronal populations in basal forebrain cholinergic neurons to generate an animal model for Alzheimer's disease. Using Flow Cytometry, this antibody has frequently been employed for panning to isolate p75NTR-expressing rat cells. MC192 has a potential use as the ligand for gene delivery into p75NTR-expressing rat cells via a receptor-mediated mechanism. FUNCTION: Low affinity receptor which can bind to NGF, BDNF, NT-3, and NT-4. Can mediate cell survival as well as cell death of neural cells. SUBUNIT: Homodimer; disulfide-linked. Interacts with p75NTR-associated cell death executor. Interacts with NGFRAP1/BEX3. Interacts with TRAF2, TRAF4, TRAF6, PTPN13 and RANBP9. Interacts through TRAF6 with SQSTM1 which bridges NGFR to NTRK1 (By similarity). Interacts with BEX1. SUBCELLULAR LOCATION: Membrane; single-pass type I membrane protein. DOMAIN: Death domain is responsible for interaction with RANBP9. PTM: N- and O-glycosylated. PTM: Phosphorylated on serine residues. SIMILARITY: Contains 1 death domain. SIMILARITY: Contains 4 TNFR-Cys repeats.

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Cyclin B1 (CCNB1) is a regulatory protein involved in mitosis. The gene product complexes with p34 (Cdk1) to form the maturation-promoting factor (MPF). Cyclin B1 contributes to the switch-like all or none behavior of the cell in deciding to commit to mitosis. Its activation is well-regulated, and positive feedback loops ensure that once the CCNB1-Cdk1 complex is activated, it is not deactivated. CCNB1-Cdk1 is involved in the early events of mitosis, such as chromosome condensation, nuclear envelope breakdown, and spindle pole assembly. Once activated, CCNB1-Cdk1 promotes several of the events of early mitosis. The active complex phosphorylates and activates 13S condensin, which helps to condense chromosomes. Another important function of the complex is to break down the nuclear envelope. The nuclear envelope is a membranous structure containing large protein complexes supported by a network of nuclear lamins. Phosphorylation of the lamins by CCBN1-Cdk1 causes them to dissociate, compromising the structural integrity of the nuclear envelope so that it breaks down. The destruction of the nuclear envelope is important because it allows the mitotic spindle to access the chromosomes. [Wiki].

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Recognises a protein of 55 kDa, which is identified as fascin-1. Its actin binding ability is regulated by phosphorylation. Antibody to fascin-1 is a very sensitive marker for Reed-Sternberg cells and variants in nodular sclerosis, mixed cellularity, and lymphocyte depletion Hodgkin's disease. It is uniformly negative in lymphoid cells, plasma cells, and myeloid cells. Fascin-1 is also expressed in dendritic cells. This marker may be helpful to distinguish between Hodgkin lymphoma and non-Hodgkin lymphoma in difficult cases. Also, the lack of expression of fascin-1 in the neoplastic follicles in follicular lymphoma may be helpful in distinguishing these lymphomas from reactive follicular hyperplasia in which the number of follicular dendritic cells is normal or increased. Antibody to fascin-1 has been suggested as a prognostic marker in neuroendocrine neoplasms of the lung as well as in ovarian cancer. Fascin-1 expression may be induced by Epstein-Barr virus (EBV) infection of B cells with the possibility that viral induction of fascin in lymphoid or other cell types must also be considered in EBV-positive cases.