Nordic BD FACS Users Meeting DNA Session. "Product update from BD Biosciences by Gerd Hempel, BD Biosciences Europe
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1 Nordic BD FACS Users Meeting DNA Session "Product update from BD Biosciences by Gerd Hempel, BD Biosciences Europe
2 DNA Transcription factors Transcription factor specific Antibodies from BD Biosciences GATA3: A Key Transcriptional Regulator in Development, Differentiation, and Cancer p53 Acetylation: A Call to Action Transcription factor specific Antibodies available from BD Biosciences
3 GATA3: A Key Transcriptional Regulator in Development, Differentiation, and Cancer is a zinc-finger transcription factor expressed during development by both hematopoietic (T cells, thymocytes) and non-hematopoietic cells (developing kidney, central and peripheral nervous system) is known to play important roles in development, differentiation, and cancer.
4 GATA3 in development Transcriptional regulation of positive selection, CD4- or CD8-lineage commitment and post-commitment development of thymocytes. Nature Reviews Immunology; Volume 9, February 2009, p
5 GATA3 in TH2-cell differentiation and function GATA-binding protein 3 is required for both STAT6-dependent and STAT6-independent pathways of T-helper-2-cell differentiation. Nature Reviews Immunology; Volume 9, February 2009, p
6 GATA3 and cancer A number of studies have been focused on the altered expression of GATA3 in various types of cancer such as breast, cervical, esophageal, and pancreatic cancer, although the precise role of GATA3 remains unclear. In breast cancer, for example, GATA3 expression is a prognostic marker of the survival outcome. High levels of GATA3 are seen in tumors of the Luminal A subtype, intermediate levels in the Luminal B subtype, and low levels in the Her2+ and Basal subtypes.
7 Tools for studying GATA3 biology GATA3 is an essential transcription factor whose biological activities affect the function of both malignant and healthy cells. An anti-gata3 monoclonal antibody (clone L ) has been co-developed in a collaboration between BD Biosciences and the National Cancer Institute as an attempt to provide tools to study these activities. Its specificity was established by Western blot analysis of mouse Th1 (2D6 cells), Th2 (D10.G4.1 cells), thymoma (EL4 cells) and B (A20 cells) cell line lysates
8 Differential expression of GATA3 by mouse Th1, Th2, thymocyte, and B cell lines Th2 Th1 T-Cell B-Cell GATA3 ~50kDa n g/ml ng/ml ng/ml Anti-actin C ng/ml Anti-actin C4
9 Expression of GATA3 by human T and B cells Jurkat Ramos Tonsil tissue Tonsil tissue 64,32,16,8,4,2,1,0.5 ng/ml 64,32,16,8,4,2,1,0.5 ng/ml GATA3 staining Isotype staining Anti-actin C4
10 Analysis of GATA3 expression by breast carcinoma cell lines MCF-7 MDA-MB-231 Pseudo-colored green: GATA3 staining Pseudo-colored blue: Hoechst 33342
11 Flow cytometric analysis of GATA3 expression by mouse and human T and B cell lines. Red: Ramos Blue: Jurkat Red: A20 (B-Cell) Blue: EL4 (T-Cell) Red: 2D6 (Th1) Blue: D10 (Th2)
12 Differential expression of GATA3 by subsets of mouse thymocytes
13 GATA3 expression in normal and malignant breast tissue GATA3 Isotype A, B: normal tissue C, D: malignant highly differentiated E, F: malignant low differentiated
14 GATA3 Antibodies
15 p53 Acetylation: A Call to Action p53 (also known as tumor protein 53 or TP53) regulates the cell cycle and is involved with preserving genomic integrity. This tumor suppressor gene is mutated in many human cancers, resulting in failed cellular responses to DNA damage. Much information is available on tumour suppressor p53 regarding the complexity of its function and regulation in carcinogenesis. Protein acetylation plays a crucial role in regulating chromatin structure and transcriptional activity. Site-specific acetylation of a growing number of non-histone proteins, including p53 and E2F, has been shown to regulate their activity, localization, specific interactions as well as stability/degradation
16 Activation of p53 and cellular response Nature Reviews Cancer; Volume 4, October 2004, p
17 p53 Acetylation: A Call to Action Gaps still exist in our knowledge regarding the role of p53 post-translational modifications in carcinogenesis and cancer prevention. A thorough understanding of p53 will be extremely useful in the development of new strategies for treating and preventing cancer, including restoration of p53 function and selective killing of tumours Monoclonal antibody L82-51 specifically recognizes p53 acetylated by cell lines or normal human peripheral blood lymphocytes that are treated with DNA-damaging agents.
18 p53 acetylation in treated HeLa cells A: untreated HeLa cells B: treated with adriamycin and trichostatin (deacetylase inhibitor)
19 p53 acetylation in treated human lymphocytes Lane 1: human PBMC untreated Lane 2-4: treated with adriamycin and trichostatin (deacetylase inhibitor) Lane 1,2: anti-p53 ack382 antibody alone Lane 3: preincubated with non-acetylated p53 K382 peptide Lane 4: preincubated with acetylated p53 ack382 peptide
20 Flow cytometric analysis of cells expressing acetylated p53 Blue: untreated Red: adriamycin/trichostatin treated
21 p53 related products from BD Biosciences
22 Transcription Marker specific Antibodies from BD Biosciences BD Biosciences features BD Pharmingen and BD Transduction Laboratories brand antibodies directed to regulatory factors the E2A family including E12, E47 and E2-2 as well as for a wide range of transcription factors including myc, Stat, NF-AT, myod, ERG, and Id. And other famos ones like NF-κB, H2AX (ps140), FoxP3, ELK-1 (ps383), and many more
23 Acknowledgement GATA3: A Key Transcriptional Regulator in Development, Differentiation, and Cancer Efthalia Chronopulou Guy Solsaa Jeanne Elia David Ernst Naomi Nobida Brad Paulsen Xiang Dong Ji Jing-Ping Shih p53 Acetylation: A Call to Action Jie Chen Cad Sisouvanthong Guo-Jian Guo David Ernst
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