SANTA CRUZ BIOTECHNOLOGY, INC. NELF-A (A-20): sc-23599

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BACKGROUND PRODUCT NELF-A, for negative elongation factor A, is a factor required for DRB- Each vial contains 200 µg IgG in 1.0 ml of PBS with < 0.1% sodium azide sensitive transcription. NELF-A is one of the five components of the NELF and 0.1% gelatin. complex that cooperates with DSIF to repress RNA polymerase II elongation. Blocking peptide available for competition studies, sc-23599 P, (100 µg pep- Control of transcription elongation requires a complex interplay between tide in 0.5 ml PBS containing < 0.1% sodium azide and 0.2% BSA). positive transcription elongation factor b (P-TEFb) and negative transcription elongation factors, DSIF and NELF. DSIF and NELF act as negative transcrip- APPLICATIONS tion elongation factors by increasing the time the polymerase spends at pause sites. DSIF/NELF inhibition of transcription is prevented by P-TEFb in NELF-A (A-20) is recommended for detection of NELF-A of mouse, rat and cooperation with FACT. NELF-A, also known as WHSC2 (Wolf-Hirschhorn human origin by Western Blotting (starting dilution 1:200, dilution range syndrome candidate 2), has a molecular mass of 57 kDa. Wolf-Hirschhorn 1:100-1:1000), immunoprecipitation [1–2 µg per 100–500 µg of total protein syndrome is a multiple malformation syndrome characterized by mental and (1 ml of cell lysate)] and immunofluorescence (starting dilution 1:50, dilution developmental defects resulting from a hemizygous deletion of the distal range 1:50-1:500). short arm of 4 (4p16.3). The human NELF-A maps to chro- Molecular Weight of NELF-A: 66 kDa. mosome 4p16.3 and encodes a 528 amino acid protein that is expressed in endothelial cells. Positive Controls: DU 145 nuclear extract: sc-24960.

REFERENCES RECOMMENDED SECONDARY REAGENTS 1. Yamaguchi, Y., Takagi, T., Wada, T., Yano, K., Furuya, A., Sugimoto, S., To ensure optimal results, the following support (secondary) reagents are Hasegawa, J., and Handa, H. 1999. NELF, a multisubunit complex containing recommended: 1) Western Blotting: use donkey anti-goat IgG-HRP: sc-2020 RD, cooperates with DSIF to repress RNA polymerase II elongation. Cell 97: (dilution range: 1:2000-1:100,000) or Cruz Marker™ compatible donkey 41-51. anti-goat IgG-HRP: sc-2033 (dilution range: 1:2000-1:5000), Cruz Marker™ Molecular Weight Standards: sc-2035, TBS Blotto A Blocking Reagent: 2. Wright, T.J., Costa, J.L., Naranjo, C., Francis-West, P., and Altherr, M.R. sc-2333 and Western Blotting Luminol Reagent: sc-2048. 2) Immunoprecip- 1999. Comparative analysis of a novel gene from the Wolf-Hirschhorn/ itation: use Protein A/G PLUS-Agarose: sc-2003 (0.5 ml agarose/ 2.0 ml). Pitt-Rogers-Danks syndrome critical region. Genomics 59: 203-212. 3) Immunofluorescence: use donkey anti-goat IgG-FITC: sc-2024 (dilution 3. Wada, T., Orphanides, G., Hasegawa, J., Kim, D.K., Shima, D., Yamaguchi, range: 1:100-1:400) or donkey anti-goat IgG-TR: sc-2783 (dilution range: Y., Fukuda, A., Hisatake, K., Oh, S., Reinberg, D., and Handa, H. 2000. 1:100-1:400) with UltraCruz™ Mounting Medium: sc-24941. FACT relieves DSIF/NELF-mediated inhibition of transcriptional elongation and reveals functional differences between P-TEFb and TFIIH. Mol. Cell 5: DATA 1067-1072. 132 K 4. Mariotti, M., Manganini, M., and Maier, J.A. 2000. Modulation of WHSC2 90 K expression in human endothelial cells. FEBS Lett. 487: 166-170. < NELF-A 55 K 5. Ping, Y.H. and Rana, T.M. 2001. DSIF and NELF interact with RNA poly- 43 K merase II elongation complex and HIV-1 Tat stimulates P-TEFb-mediated phosphorylation of RNA polymerase II and DSIF during transcription elon- 34 K gation. J. Biol. Chem. 276: 12951-12958.

6. Renner, D.B., Yamaguchi, Y., Wada, T., Handa, H., and Price, D.H. 2001. NELF-A (A-20): sc-23599. Western blot analysis of A highly purified RNA polymerase II elongation control system. J. Biol. NELF-A expression in DU 145 nuclear extract. Chem. 276: 42601-42609. 7. Online Mendelian Inheritance in Man, OMIM™. 2001. Johns Hopkins RESEARCH USE University, Baltimore, MD. MIM Number: 606026. World Wide Web URL: For research use only, not for use in diagnostic procedures. http://www.ncbi.nlm.nih.gov/omim/ PROTOCOLS SOURCE See our web site at www.scbt.com or our catalog for detailed protocols NELF-A (A-20) is an affinity purified goat polyclonal antibody raised against and support products. a peptide mapping at the N-terminus of NELF-A of human origin.

STORAGE Store at 4° C, **DO NOT FREEZE**. Stable for one year from the date of shipment. Non-hazardous. No MSDS required.

Santa Cruz Biotechnology, Inc. 1.800.457.3801 831.457.3800 fax 831.457.3801 Europe +00800 4573 8000 49 6221 4503 0 www.scbt.com