Datasheet for Histone H3.2 Human, Recombinant (M2506; Lot 0021608)
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encoding the cloned human histone H3.2 gene, Histone H3.2 Human, Recombinant is reaction containing 10 µg of Histone H3.2 Human, Histone H3.2 HIST2H3A or HIST2H3C. (Genbank accession 15257.09 Da as determined by ESI-TOF MS Recombinant with 1 µg of a mixture of single and number: BC130637) (Electrospray Ionization-Time of Flight Mass double-stranded [3H] E. coli DNA (200,000 cpm/ Human, Recombinant Spectrometry). The average mass calculated from µg) for 4 hours at 37°C released < 0.1% of the Supplied in: 20 mM Sodium Phosphate (pH 7.0), primary sequence is 15256.82 Da. This confirms total radioactivity. 1-800-632-7799 300 mM NaCl, 1 mM EDTA and 1 mM DTT. the protein identity of the histone. For a typical [email protected] example of mass spectrometry data, please see Endonuclease Assay: Incubation of a 50 µl www.neb.com Note: The protein concentration (1 mg/ml, 66 µM) the product page at www.neb.com. reaction containing 10 µg of Histone H3.2 M2506S 002160818081 is calculated using the molar extinction coefficient Human, Recombinant with 1 µg of φX174 RF I for Histone H3.2 (3960) and its absorbance at N-terminal Protein Sequencing: Protein identity (supercoiled) plasmid DNA for 4 hours at 37°C 280 nm (4,5). 1.0 A units = 3.9 mg/ml was confirmed using Edman Degradation to resulted in < 5.0% conversion to RF II form M2506S 280 sequence the intact protein. (nicked circle) as determined by agarose gel 100 µg 1.0 mg/ml Lot: 0021608 Synonym: Histone H3/m, H3/o electrophoresis. Enzyme Modification: SET7 Methyltransferase: RECOMBINANT Store at –20°C Exp: 8/18 Gene Synonym: H3F2, H3FM After incubation of a 25 µl reaction for 10 minutes Protein Sequence: ARTKQTARKSTGGKAPRKQLA Description: Histone H3 combines with Histone at 37°C, 1 unit of SET7 methyltransferase TKAARKSAPATGGVKKPHRYRPGTVALREIRRYQKS H4 to form the H3/H4 tetramer. Two H2A/H2B Quality Control Assays: transfers 20 pmols of methyl group to Histone TELLIRKLPFQRLVREIAQDFKTDLRFQSSAVMALQEA heterodimers interact with an H3/H4 tetramer to SDS-PAGE: 0.5, 1.0, 2.0, 5.0, 10.0 µg of Histone H3.2 Human, Recombinant. SEAYLVGLFEDTNLCAIHAKRVTIMPKDIQLARRIRGE form the histone octamer (1,2). It is also modified H3.2 Human, Recombinant were loaded on a 10– RA (Genbank accession number: Q71DI3) Protease Assay: After incubation of 10 µg by various enzymes and can act as a substrate for 20% Tris-Glycine SDS-PAGE gel and stained with of Histone H3.2 Human, Recombinant with a them. These modifications have been shown to be Coomassie Blue. The calculated molecular weight References: standard mixture of proteins for 4 hours at 37°C, important in gene regulation. is 15256.82 Da. Its apparent molecular weight on 10–20% Tris-Glycine SDS-PAGE gel is ~17 kDa. no proteolytic activity could be detected by SDS- (see other side) Histone H3.2, an H3 variant that is found in all For a typical example of gel image, please see the PAGE. eukaryotes except budding yeast, is replication de- product page at www.neb.com. pendent and is associated with gene silencing (3). Exonuclease Assay: Incubation of a 50 µl Mass Spectrometry: The mass of purified Source: An E. coli strain that carries a plasmid CERTIFICATE OF ANALYSIS encoding the cloned human histone H3.2 gene, Histone H3.2 Human, Recombinant is reaction containing 10 µg of Histone H3.2 Human, Histone H3.2 HIST2H3A or HIST2H3C. (Genbank accession 15257.09 Da as determined by ESI-TOF MS Recombinant with 1 µg of a mixture of single and number: BC130637) (Electrospray Ionization-Time of Flight Mass double-stranded [3H] E. coli DNA (200,000 cpm/ Human, Recombinant Spectrometry). The average mass calculated from µg) for 4 hours at 37°C released < 0.1% of the Supplied in: 20 mM Sodium Phosphate (pH 7.0), primary sequence is 15256.82 Da. This confirms total radioactivity. 1-800-632-7799 300 mM NaCl, 1 mM EDTA and 1 mM DTT. the protein identity of the histone. For a typical [email protected] example of mass spectrometry data, please see Endonuclease Assay: Incubation of a 50 µl www.neb.com Note: The protein concentration (1 mg/ml, 66 µM) the product page at www.neb.com. reaction containing 10 µg of Histone H3.2 M2506S 002160818081 is calculated using the molar extinction coefficient Human, Recombinant with 1 µg of φX174 RF I for Histone H3.2 (3960) and its absorbance at N-terminal Protein Sequencing: Protein identity (supercoiled) plasmid DNA for 4 hours at 37°C 280 nm (4,5). 1.0 A units = 3.9 mg/ml was confirmed using Edman Degradation to resulted in < 5.0% conversion to RF II form M2506S 280 sequence the intact protein. (nicked circle) as determined by agarose gel 100 µg 1.0 mg/ml Lot: 0021608 Synonym: Histone H3/m, H3/o electrophoresis. Enzyme Modification: SET7 Methyltransferase: RECOMBINANT Store at –20°C Exp: 4/18 Gene Synonym: H3F2, H3FM After incubation of a 25 µl reaction for 10 minutes Protein Sequence: ARTKQTARKSTGGKAPRKQLA Description: Histone H3 combines with Histone at 37°C, 1 unit of SET7 methyltransferase TKAARKSAPATGGVKKPHRYRPGTVALREIRRYQKS H4 to form the H3/H4 tetramer. Two H2A/H2B Quality Control Assays: transfers 20 pmols of methyl group to Histone TELLIRKLPFQRLVREIAQDFKTDLRFQSSAVMALQEA heterodimers interact with an H3/H4 tetramer to SDS-PAGE: 0.5, 1.0, 2.0, 5.0, 10.0 µg of Histone H3.2 Human, Recombinant. SEAYLVGLFEDTNLCAIHAKRVTIMPKDIQLARRIRGE form the histone octamer (1,2). It is also modified H3.2 Human, Recombinant were loaded on a 10– RA (Genbank accession number: Q71DI3) Protease Assay: After incubation of 10 µg by various enzymes and can act as a substrate for 20% Tris-Glycine SDS-PAGE gel and stained with of Histone H3.2 Human, Recombinant with a them. These modifications have been shown to be Coomassie Blue. The calculated molecular weight standard mixture of proteins for 4 hours at 37°C, important in gene regulation. is 15256.82 Da. Its apparent molecular weight on 10–20% Tris-Glycine SDS-PAGE gel is ~17 kDa. no proteolytic activity could be detected by SDS- (see other side) Histone H3.2, an H3 variant that is found in all For a typical example of gel image, please see the PAGE. eukaryotes except budding yeast, is replication de- product page at www.neb.com. pendent and is associated with gene silencing (3). Exonuclease Assay: Incubation of a 50 µl Mass Spectrometry: The mass of purified Source: An E. coli strain that carries a plasmid CERTIFICATE OF ANALYSIS 1. Kornberg, R.D. (1977) Annu. Rev. Biochem. 46, 931–954. ISO 9001 ISO 14001 ISO 13485 2. van Holde, K.E. (1989) Chromatin, 1–497. Registered Registered Registered Quality Environmental Medical Devices 3. Hake, S.B. et al (2006) J.Biol. Chem. 281, 559- Management Management 568. NEW ENGLAND BIOLABS® is a registered trademark of New England 4. Gill, S.C. and von Hippel, P.H. (1989) Anal. Bio- Biolabs, Inc. chem. 182, 319–326. This product is intended for research purposes only. This product 5. Pace, C.N. et al. (1995) Protein Science, 4, is not intended to be used for therapeutic or diagnostic purposes in 2411–2423. humans or animals. Page 2 (M2506) References: 1. Kornberg, R.D. (1977) Annu. Rev. Biochem. 46, ISO 9001 ISO 14001 ISO 13485 931–954. Registered Registered Registered Quality Environmental Medical Devices 2. van Holde, K.E. (1989) Chromatin, 1–497. Management Management 3. Hake, S.B. et al (2006) J.Biol. Chem. 281, 559- NEW ENGLAND BIOLABS® is a registered trademark of New England 568. Biolabs, Inc. 4. Gill, S.C. and von Hippel, P.H. (1989) Anal. Bio- This product is intended for research purposes only. This product chem. 182, 319–326. is not intended to be used for therapeutic or diagnostic purposes in 5. Pace, C.N. et al. (1995) Protein Science, 4, humans or animals. 2411–2423. Page 2 (M2506).