Source: An E. coli strain that carries a plasmid Mass Spectrometry: The mass of purified double-stranded [3H] E. coli DNA (200,000 cpm/ encoding the cloned human histone H3.2 gene, Histone H3.2 Human, Recombinant is 15257.09 Da µg) for 4 hours at 37°C released < 0.1% of the total Histone H3.2 HIST2H3A or HIST2H3C. (Genbank accession as determined by ESI-TOF MS (Electrospray radioactivity. Human, Recombinant number: BC130637) Ionization-Time of Flight Mass Spectrometry). The average mass calculated from primary sequence Endonuclease Assay: Incubation of a 50 µl 1-800-632-7799 Supplied in: 20 mM Sodium Phosphate (pH 7.0), is 15256.82 Da. This confirms the protein identity reaction containing 10 µg of Histone H3.2 Human, [email protected] 300 mM NaCl, 1 mM EDTA and 1 mM DTT. as well as the absence of any modifications of the Recombinant with 1 µg of φX174 RF I (supercoiled) www.neb.com histone. For a typical example of mass spectrometry plasmid DNA for 4 hours at 37°C resulted in M2506S 002131215121 Note: The protein concentration (1 mg/ml, 66 µM) data, please see the product page at www.neb.com. < 5.0% conversion to RF II form (nicked circle) as is calculated using the molar extinction coefficient determined by agarose gel electrophoresis. M2506S B r for Histone H3.2 (3960) and its absorbance at N-terminal Protein Sequencing: Protein identity was confirmed using Edman Degradation to sequence the Protein Sequence: ARTKQTARKSTGGKAPRKQLA 100 µg 1.0 mg/ml Lot: 0021312 280 nm (4,5). 1.0 A280 units = 3.9 mg/ml intact protein. TKAARKSAPATGGVKKPHRYRPGTVALREIRRYQKS RECOMBINANT Store at –20°C Exp: 12/15 Synonym: Histone H3/m, H3/o TELLIRKLPFQRLVREIAQDFKTDLRFQSSAVMALQEA Enzyme Modification: SET7 Methyltransferase: SEAYLVGLFEDTNLCAIHAKRVTIMPKDIQLARRIRGE Description: Histone H3 combines with Histone H4 Gene Synonym: H3F2, H3FM After incubation of a 25 µl reaction for 10 minutes RA (Genbank accession number: Q71DI3) to form the H3/H4 tetramer. Two H2A/H2B heterodi- at 37°C, 1 unit of SET7 methyltransferase transfers mers interact with an H3/H4 tetramer to form the Quality Control Assays: 20 pmols of methyl group to Histone H3.2 Human, References: histone octamer (1,2). It is also modified by various SDS-PAGE: 0.5, 1.0, 2.0, 5.0, 10.0 µg of Histone Recombinant. 1. Kornberg, R.D. (1977) Annu. Rev. Biochem. 46, enzymes and can act as a substrate for them. These H3.2 Human, Recombinant were loaded on a 931–954. modifications have been shown to be important in 10–20% Tris-Glycine SDS-PAGE gel and stained Protease Assay: After incubation of 10 µg of Histone 2. van Holde, K.E. (1989) Chromatin, 1–497. gene regulation. with Coomassie Blue. The calculated molecular H3.2 Human, Recombinant with a standard mixture 3. Hake, S.B. et al (2006) J.Biol. Chem. 281, 559- of proteins for 4 hours at 37°C, no proteolytic activity 568. Histone H3.2, an H3 variant that is found in all weight is 15256.82 Da. Its apparent molecular could be detected by SDS-PAGE. 4. Gill, S.C. and von Hippel, P.H. (1989) Anal. eukaryotes except budding yeast, is replication de- weight on 10–20% Tris-Glycine SDS-PAGE gel is ~17 kDa. For a typical example of gel image, please Biochem. 182, 319–326. pendent and is associated with gene silencing (3). Exonuclease Assay: Incubation of a 50 µl see the product page at www.neb.com. 5. Pace, C.N. et al. (1995) Protein Science, 4, reaction containing 10 µg of Histone H3.2 Human, 2411–2423. Recombinant with 1 µg of a mixture of single and CERTIFICATE OF ANALYSIS Source: An E. coli strain that carries a plasmid Mass Spectrometry: The mass of purified double-stranded [3H] E. coli DNA (200,000 cpm/ encoding the cloned human histone H3.2 gene, Histone H3.2 Human, Recombinant is 15257.09 Da µg) for 4 hours at 37°C released < 0.1% of the total Histone H3.2 HIST2H3A or HIST2H3C. (Genbank accession as determined by ESI-TOF MS (Electrospray radioactivity. Human, Recombinant number: BC130637) Ionization-Time of Flight Mass Spectrometry). The average mass calculated from primary sequence Endonuclease Assay: Incubation of a 50 µl 1-800-632-7799 Supplied in: 20 mM Sodium Phosphate (pH 7.0), is 15256.82 Da. This confirms the protein identity reaction containing 10 µg of Histone H3.2 Human, [email protected] 300 mM NaCl, 1 mM EDTA and 1 mM DTT. as well as the absence of any modifications of the Recombinant with 1 µg of φX174 RF I (supercoiled) www.neb.com histone. For a typical example of mass spectrometry plasmid DNA for 4 hours at 37°C resulted in M2506S 002131215121 Note: The protein concentration (1 mg/ml, 66 µM) data, please see the product page at www.neb.com. < 5.0% conversion to RF II form (nicked circle) as is calculated using the molar extinction coefficient determined by agarose gel electrophoresis. M2506S B r for Histone H3.2 (3960) and its absorbance at N-terminal Protein Sequencing: Protein identity was confirmed using Edman Degradation to sequence the Protein Sequence: ARTKQTARKSTGGKAPRKQLA 100 µg 1.0 mg/ml Lot: 0021312 280 nm (4,5). 1.0 A280 units = 3.9 mg/ml intact protein. TKAARKSAPATGGVKKPHRYRPGTVALREIRRYQKS RECOMBINANT Store at –20°C Exp: 12/15 Synonym: Histone H3/m, H3/o TELLIRKLPFQRLVREIAQDFKTDLRFQSSAVMALQEA Enzyme Modification: SET7 Methyltransferase: SEAYLVGLFEDTNLCAIHAKRVTIMPKDIQLARRIRGE Description: Histone H3 combines with Histone H4 Gene Synonym: H3F2, H3FM After incubation of a 25 µl reaction for 10 minutes RA (Genbank accession number: Q71DI3) to form the H3/H4 tetramer. Two H2A/H2B heterodi- at 37°C, 1 unit of SET7 methyltransferase transfers mers interact with an H3/H4 tetramer to form the Quality Control Assays: 20 pmols of methyl group to Histone H3.2 Human, References: histone octamer (1,2). It is also modified by various SDS-PAGE: 0.5, 1.0, 2.0, 5.0, 10.0 µg of Histone Recombinant. 1. Kornberg, R.D. (1977) Annu. Rev. Biochem. 46, enzymes and can act as a substrate for them. These H3.2 Human, Recombinant were loaded on a 931–954. modifications have been shown to be important in 10–20% Tris-Glycine SDS-PAGE gel and stained Protease Assay: After incubation of 10 µg of Histone 2. van Holde, K.E. (1989) Chromatin, 1–497. gene regulation. with Coomassie Blue. The calculated molecular H3.2 Human, Recombinant with a standard mixture 3. Hake, S.B. et al (2006) J.Biol. Chem. 281, 559- of proteins for 4 hours at 37°C, no proteolytic activity 568. Histone H3.2, an H3 variant that is found in all weight is 15256.82 Da. Its apparent molecular could be detected by SDS-PAGE. 4. Gill, S.C. and von Hippel, P.H. (1989) Anal. eukaryotes except budding yeast, is replication de- weight on 10–20% Tris-Glycine SDS-PAGE gel is ~17 kDa. For a typical example of gel image, please Biochem. 182, 319–326. pendent and is associated with gene silencing (3). Exonuclease Assay: Incubation of a 50 µl see the product page at www.neb.com. 5. Pace, C.N. et al. (1995) Protein Science, 4, reaction containing 10 µg of Histone H3.2 Human, 2411–2423. Recombinant with 1 µg of a mixture of single and CERTIFICATE OF ANALYSIS.