HeLa Nuclear Extracts
€29,00 – €595,00
Volume:from 150ul to 15ml. Concentration:~6mg/ml (Bradford). Hela Nuclear Extracts are prepared from fresh cells in exponential phase only (Viability>95%).
The HeLa cell line was established from an adenocarcinoma of the cervix in 1952. It is the first continuous human cell line.
Cultures are screened for the presence of bacteria, yeast, fungi and mycoplasma (DNA amplification). NBCS used in the culture medium is certified from New Zealand origin.
HeLa Nuclear Extracts Production
HeLa cells are grown in sonoperfused fedbatch (cytostat) mode at a constant concentration of 5×106 cells/ml (cell viability: 93%-99%) under GLP conditions in our facility in Mons, Belgium. Cells are harvested in exponential phase.
The Nuclear Extracts are prepared according to:
Dignam, J. D., Lebovitz, R. M., and Roeder, R. G. (1983) Nucleic Acids Res. 11, 1475-1489
Conc. Protein (Bradford) = +/- 6 mg/ml
Buffer used for dialysis of our Nuclear Extracts is:
- HEPES 20mM
- KCl 100mM
- EDTA 0.2mM
- Glycerol 20%
- PMSF 0.2mM
- DTT 0.5mM
Our HeLa Nuclear Extracts are used by research or production entities worldwide for the study of biochemical processing, high throughput screening or purification of biological material from human origin.
Source of HDAC activity, transcription factors, chromatin proteins and histones.
Also suitable for use in:
- separation by SDS-PAGE;
- Gel Shift assays used for protein-DNA interactions studies;
- HDAC assays;
- positive control for Western Blot assays;
- in vitro splicing;
- transcription factors studies;
- cell division cycle and apoptosis studies;
- spliceosome and other proteome studies (DNA ends, snRNP’s, DNA PKcs, …).
Downloads – Documents
- Laggerbauer, B., Lauber, J. and Lührmann, R. (1996). Identification of an RNA-dependent ATPase activity in mammalian U5 SnRNPs. Nucl. Ac. Res. 24, 868-875.
- Sirand-Pugnet, P., Durosay, P., Clouet d’Orval, B., Brody, E., Marie, J. (1995). R-Tropomyosin pre-mRNA folding around a muscle-specific exon interferes with several steps of spliceosome assembly. J. Mol. Biol. 251, 591-602.
- Gell, D. and Jackson, S. P. (1999). Mapping of protein-protein interactions within the DNA-dependent protein kinase complex. Nucl. Ac. Res. 27, 3494-3502.
- Izzard, R.A., Jackson, S.P. and Smith, G.C.M. (1999). Competitive and non-competitive inhibition of the DNA-dependent protein kinase. Cancer Res. 59, 2581-2586.
- Lakin, N.D., Hann, B.C. and Jackson, S.P. (1999) The ataxia-telangiectasia related protein ATR mediates DNA-dependent phosphorylation of P53. Oncogene 68, 3989-3995.
- Smith, G.C., Cary, R.B., Lakin, N.D., Hann, B.C., Teo, S._H., Chen, D.J. and Jackson, S.P. (1999) Purification and DNA binding properties of the ataxia-telangiectasia gene product ATM. Proc. Natl. Acad. Sci. USA 96, 11134-11139.
- Smith, G.C., d’Adda di Fagagna, F., Lakin, N.D. and Jackson, S.P. (1999) Cleavage and inactivation of ATM during apoptosis. Molec. Cell. Biol. 19, 6076-6084.
- Chew, S.L., Baginsky, L., Eperon, I.C. (2000) An exonic splicing silencer in the testes-specific DNA ligase III B exon. Nucleic Acid Res. 28, 402-410.
- Deplus, R., Brenner, C., Burgers, W.A., Putmans, P., Kouzarides, T., de Launoit, Y., Fuks, F. (2002) Dnmt3L is a transcriptional repressor that recruits histone deacetylase. Nucleic Acid Res. 30, n°17, 3831-3838.
- Hallay, H., Locker, N., Ayadi, L., Ropers, D., Guittet, E. and Branlant, C. (2006) Biochemical and NMR study on the competition between proteins SC35, SRp40 and hnRNP A1 at the HIV-1 Tat exon 2 splicing site. J. Biol. Chem., Vol. 281, Issue 48, 37159-37174
- Hanna, C., Gjerde, D., Nguyen, L., Dickman, M., Brown, P. and Hornby, D. (2006) Micro-scale open-tube capillary separations of functional proteins. Analytical Biochemistry 350, 128-137.
- Jacquenet, S., Méreau, A., Bilodeau, P.S., Damier, L., Martin Stoltzfus, C. and Branlant, C. (2001) A Second Exon Splicing Silencer within Human Immunodeficiency Virus Type 1 tat Exon 2 Represses Splicing of Tat mRNA and Binds Protein hnRNP H*. J. Biol. Chem., 276, No 44, Issue of November 2, 40464-40475
- Roche,K.C., Wiechens,N., Owen-Hughes,T and Perkins, N.D. (2004) The FHA domain protein SNIP1 is a regulator of the cell cycle and cyclin D1 expression. Oncogene 23, 8185-8195
- Brandt, D., Marion, S., Griffiths, G., Watanabe, T., Kaibuchi, K. and Grosse, R. (2007) Dia 1 and IQGAP1 interact in cell migration and phagocytic cup formation. Journal of Cell Biology 178, 193-200
- Ropers, D., Ayadi, L., Gattoni, R., Jacquenet, S., Damier, L., Branlant, C. and Stévenin, J. (2004) Differential effects of the SR proteins 9G8, SC35, ASF/SF2 and SRp40 on the utilization of the A1 to A5 splicing sites of HIV-1 RNA. J. Biol. Chem., 279, 29963-29973
- Gehring, N.H., Lamprinaki, S., Kulozik, A.E. and Hentze, M. (2009) Disassembly of exon junction complexes by PYM. Cell 137, 536-548
- Gehring, N.H., Lamprinaki, S., Hentze, M. and Kulozik, A.E. (2009) The hierarchy of exon-junction complex assembly by the spliceosome explains key features of mammalian nonsense-mediated mRNA decay. Plos Biology 7 (5) e 1000120
- Rivera-Calzada, A., Spagnolo, L., Laurence H., P., Llorca, O. (2007) Structural model of full-lengh human Ku70-Ku80 heterodimer and its recognition of DNA and DNA-PKcs. EMBO reports 8, 56-62
- Stefan Reber, Jolanda Stettler1, Giuseppe Filosa, Martino Colombo Daniel Jutzi, Silvia C Lenzken, Christoph Schweingruber, Rémy Bruggmann, Angela Bachi, Silvia ML Barabino, Oliver Mühlemann & Marc-David Ruepp (2016) Minor intron splicing is regulated by FUS and affected by ALS-associated FUS mutants. The EMBO Journal 35: 1504–1521
- JiwenYang, Lee-HsuehHung, ThomasLicht, SawaKostin, MarioLooso, EkaterinaKhrameeva, AlbrechtBindereif, AndreSchneider, ThomasBraun. (2014) RBM24 Is a Major Regulator of Muscle-Specific Alternative Splicing. Developmental Cell Volume 31, Issue 1, 13 October 2014, Pages 87-99
- Vivien Berthelot, Gildas Mouta-Cardoso, Nadia Hégarat, François Guillonneau, Jean-Christophe François, Carine Giovannangeli, Danièle Praseuth, Filippo Rusconi. (2016) The human DNA ends proteome uncovers an unexpected entanglement of functional pathways. Nucleic Acids Research, Volume 44, Issue 10, 2 June 2016, Pages 4721–4733
- Katarzyna Dorota Raczynska Marc-David Ruepp Aleksandra Brzek Stefan Reber Valentina Romeo Barbara Rindlisbacher Manfred Heller Zofia Szweykowska-Kulinska Artur Jarmolowski Daniel Schümperli. (2015) FUS/TLS contributes to replication-dependent histone gene expression by interaction with U7 snRNPs and histone-specific transcription factors. Nucleic Acids Research, Volume 43, Issue 20, 16 November 2015, Pages 9711–9728
- Tim Schneider, Lee-Hsueh Hung, Silke Schreiner, Stefan Starke, Heinrich Eckhof , Oliver Rossbach, Stefan Reich, Jan Medenbach, and Albrecht Bindereif. (2016) CircRNA-protein complexes: IMP3 protein component defines subfamily of circRNPs. Nature Sci Rep. 2016; 6: 31313.
- Andri Christodoulou and Hideki Yokoyama. (2015) Purification of nuclear localization signal-containing proteins and its application to investigation of the mechanisms of the cell division cycle. Small GTPases. 2015 Jan-Mar; 6(1): 20–27
- Lorenzo Lafranchi, Harmen R de Boer, Elisabeth GE de Vries, Shao-En Ong, Alessandro A Sartori and Marcel ATM van Vugt. (2014) APC/CCdh1 controls CtIP stability during the cell cycle and in response to DNA damage. EMBO J. 2014 Dec 1; 33(23): 2860–2879.
- Takashi Ochi, Andrew N. Blackford, Julia Coates, Satpal Jhujh, Shahid Mehmood, Naoka Tamura, Jon Travers, Qian Wu, Viji M. Draviam, Carol V. Robinson, Tom L. Blundell, and Stephen P. Jackson (2015) PAXX, a paralog of XRCC4 and XLF, interacts with Ku to promote DNA double-strand break repair. Science. 2015 Jan 9; 347(6218): 185–188.
- Andrew N. Blackford, Jadwiga Nieminuszczy, Rebekka A. Schwab, Yaron Galanty, Stephen P. Jackson, and Wojciech Niedzwiedz (2015) TopBP1 Interacts with BLM to Maintain Genome Stability but Is Dispensable for Preventing BLM Degradation. Mol Cell. 2015 Mar 19; 57(6): 1133–1141
- Pia-Amata Leimbacher, Samuel E.Jones, Ann-Marie K.Shorrocks, Marade Marco Zompit, Matthew Day, Jordy Blaauwendraad, Diana Bundschuh, Sarah Bonham, Roman Fischer, Daniel Fink, Benedikt M.Kessler, Antony W.Oliver, Laurence H.Pearl, Andrew N.Blackford, ManuelStucki (2019) MDC1 Interacts with TOPBP1 to Maintain Chromosomal Stability during Mitosis. Mol Cell. 2019 May 2; 74(3): 571-583.e8