AM26558BT-N OriGene Technologies Inc. OriGene EU Acris Antibodies GmbH 9620 Medical Center Drive, Ste 200 Schillerstr. 5 Rockville, MD 20850 32052 Herford UNITED STATES GERMANY Phone: +1-888-267-4436 Phone: +49-5221-34606-0 Fax: +1-301-340-8606 Fax: +49-5221-34606-11 [email protected] [email protected] Monoclonal Antibody to TCL1A / TCL1 - Biotin Alternate names: T-cell leukemia/lymphoma protein 1A, TCL-1 Catalog No.: AM26558BT-N Quantity: 0.1 ml Concentration: 0.5 mg/ml Background: TCL1 (T cell leukemia/lymphoma 1), MTCP1 (mature T cell proliferation 1) and TCL1b belong to the TCL1 proto-oncogene family. The TCL1 gene at chromosome 14q32.1 is co mmonly activated in T cell neoplasms by chromosome r earrangements such as inversions inv(14)(q11;q32) and translocations t(14;14)(q11;q32) or t(7;14)(q35;q32). TCL1 expression is limited to immature thymocytes in early T-cell progenitors and from pre-B to mature B cells in the B-cell lineage. The TCL1/MTCP1/TCL1b proto-oncogene activation is the hallmark of human T-cell prolymphocytic leukemia (T-PLL), a form of adult leukemia. In addition to T- PLL, TCL1 is overexpressed in Burkitt's lymphoma cell lines, the majority of AIDS-related non-Hodgkin’s lymphoma-designated immunoblastic lymphoma plasmacytoids, lymphoblastic lymphoma, chronic lymphocytic leukemia, mantle cell lymphoma, follicular lymphoma, diffuse large B-cell lymphoma, and primary cutaneous B-cell lymphoma. It is also implicated in the development of hematopoietic abnormalities in patients with ataxia- telangiectasia. The members of the TCL1 proto-oncogene family bind to the serine/threonine kinase Akt (PKB), increasing its phosphorylation status and kinase activity. Akt is a crucial regulator in transduction of antiapoptotic and proliferative signals in T cells. TCL1 facilitates heterodimerization of different Akt molecules, which may contribute to the development of human malignancies associated w ith TCL1 overexpression. Uniprot ID: P56279 NCBI: 9606 GeneID: 8115 Host / Isotype: Mouse / IgG1 Recommended SM10B (for use in human samples) Isotype Controls: Clone: 27D6/21 Immunogen: Recombinant human TCL1 Format: State: Liquid Ig fraction Purification: Protein A agarose Label: Biotin Applications: Flow cytometry: 5-10 µg/ml (final concentration). For details see protocol below. Not recommended for Immunohistochemistry. For research and in vitro use only. Not for diagnostic or therapeutic work. OG/20130815 Material Safety Datasheets are available at www.acris-antibodies.com or on request. Acris Antibodies is now part of the OriGene family. Learn more at www.origene.com 1 / 2 AM26558BT-N: Monoclonal Antibody to TCL1A / TCL1 - Biotin Other applications not tested. Optimal dilutions are dependent on conditions and should be determined by the user. Specificity: This antibody reacts with TCL1. Clone 27D6/20 does not cross-react with TCL1b and MTCP1 (ref 7). Species Reactivity: Tested: Human Add. Information: This product was originally produced by MBL International. Storage: Store at 2-8 °C. Shelf life: one year from despatch. General Readings: 1. Boccellato, F., et al., J. Virol. 81, 2274-2282 (2007). 2. Vermi, W., et al., Blood 107 , 454-462 (2006). 3. Zanesi, N., et al., Cancer Res. 66 , 915-920 (2006). 4. Künstle, G., et al., Mol. Cell. Biol. 22 , 1513-1525 (2002). 5. Laine, J., et al., J. Biol. Chem. 277, 3743-3751 (2002). 6. Bichi, R., et al., PNAS 99, 6955-6960 (2002). 7. Narducci, M. G., et al., Cancer Res. 60 , 2095-2100 (2000). Protocols: Flow cytometric analysis for floating cells We usually use Fisher tubes or equivalents as reaction tubes for all steps described below. 1) Wash the cells 3 times with washing buffer [PBS containing 2% fetal calf serum (FCS) and 0.1% NaN3]. 2) Add 100 µ L of 4% paraformaldehyde (PFA) in PBS to the cell pellet after tapping. Mix well, then fix the cells for 10 minutes at 4 o C. 3) Wash the cells 3 times with washing buffer. 4) Add 100 µ L of 0.1% Saponin in PBS to the cell pellet after tapping. Mix well, then permeabilize the cells for 10 minutes at 4 o C. 5) Wash the cells 3 times with washing buffer. 6) Add 20 µ L of Clear Back (human Fc receptor blocking reagent) to the cell pellet after tapping. Mix well and incubate for 5 minutes at room temperature. 7) Add 20 µ L of the primary antibody at the concentration as suggest in the APPLICATIONS diluted in the washing buffer. Mix well and incubate for 20 minutes at room temperature. 8) Add 1 mL of the washing buffer followed by centrifugation at 500 x g for 1 minute at room temperature. Remove supernatant by careful aspiration. 9) Add 20 µ L of 1:100 PE conjugated streptavidin diluted with the washing buffer. Mix well and incubate for 20 minut es at room temperature. 10) Add 1 mL of the washing buffer followed by centrifugation at 500 x g for 1 minute at room temperature. Remove supernatant by careful aspiration. 11) Resuspend the cells with 500 µ L of the washing buffer and analyze by a flow cytometer. (Positive control for Flow cytometry; Raji) Pictures: Flow cytometric analysis of TCL1 expression on Raji. Open histogram indicates the reaction of isotypic control to the cells. Shaded histogram indicates the reaction of AM26558BT-N to the cells. For research and in vitro use only. Not for diagnostic or therapeutic work. OG/20130815 Material Safety Datasheets are available at www.acris-antibodies.com or on request. Acris Antibodies is now part of the OriGene family. Learn more at www.origene.com 2 / 2 Powered by TCPDF (www.tcpdf.org).