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Datasheet for Histone H2B Human, Recombinant (M2505; Lot 0031404)

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Datasheet for Histone H2B Human, Recombinant (M2505; Lot 0031404) Supplied in: 20 mM Sodium Phosphate (pH 7.0), Quality Control Assays: Ionization-Time of Flight Mass Spectrometry). The Histone H2B 300 mM NaCl and 1 mM EDTA. SDS-PAGE: 0.5 µg, 1.0 µg, 2.0 µg, 5.0 µg, average mass calculated from primary sequence Human, Recombinant 10.0 µg Histone H2B Human, Recombinant were is 13788.97 Da. This confirms the protein identity Note: The protein concentration (1 mg/ml, 73 µM) loaded on a 10–20% Tris-Glycine SDS-PAGE gel as well as the absence of any modifications of the is calculated using the molar extinction coefficient and stained with Coomassie Blue. The calculated histone. 1-800-632-7799 for Histone H2B (6400) and its absorbance at molecular weight is 13788.97 Da. Its apparent [email protected] 280 nm (3,4). 1.0 A units = 2.2 mg/ml N-terminal Protein Sequencing: Protein identity 280 molecular weight on 10–20% Tris-Glycine SDS- www.neb.com was confirmed using Edman Degradation to PAGE gel is ~17 kDa. M2505S 003140416041 Synonyms: Histone H2B/q, Histone H2B.1, sequence the intact protein. Histone H2B-GL105 Mass Spectrometry: The mass of purified Histone H2B Human, Recombinant is 13788.5 Da Protease Assay: After incubation of 10 µg of M2505S B r kDa 1 2 3 4 5 6 7 as determined by ESI-TOF MS (Electrospray Histone H2B Human, Recombinant with a standard 250 4.0 mixture of proteins for 2 hours at 37°C, no 100 µg 1.0 mg/ml Lot: 0031404 150 13788.5 100 proteolytic activity could be detected by SDS- RECOMBINANT Store at –20°C Exp: 4/16 80 PAGE. 60 Description: Histone H2B combines with Histone 50 3.0 ) 6 Exonuclease Assay: Incubation of a 50 µl H2A to form the H2A-H2B heterodimer. Two H2A/ 40 30 reaction containing 10 µg of Histone H2B Human, H2B heterodimers interact with an H3/H4 tetramer 25 Recombinant with 1 µg of a mixture of single and to form the histone octamer. (1,2) Histone H2B is 2.0 3 , Counts (x10 double-stranded [ H] E. coli DNA (200,000 cpm/ also modified by various enzymes and can act as 20 15 µg) for 4 hours at 37°C released < 0.1% of the total a substrate for them. These modifications have Intensity 10 radioactivity. been shown to be important in gene regulation. 1.0 SDS-PAGE analysis of Histone H2B Human, Recombinant. Endonuclease Assay: Incubation of a 50 µl Source: An E.coli strain that carries a plasmid Lane 1 and 7: NEB Protein Ladder (NEB #P7703), Lane 2-6: reaction containing 10 µg of Histone H2B encoding the cloned human histone H2B gene, 0.5 µg, 1.0 µg, 2.0 µg, 5.0 µg, 10.0 µg Histone H2B Human, Recombinant. 0.0 Human, Recombinant with 1 µg of φX174 RF I HIST2H2BE or H2BFQ. (Genbank accession 10000 12000 14000 16000 18000 Mass (Da) number: AY131979) (See other side) ESI-TOF Analysis of Histone H2B Human, Recombinant. CERTIFICATE OF ANALYSIS Supplied in: 20 mM Sodium Phosphate (pH 7.0), Quality Control Assays: Ionization-Time of Flight Mass Spectrometry). The Histone H2B 300 mM NaCl and 1 mM EDTA. SDS-PAGE: 0.5 µg, 1.0 µg, 2.0 µg, 5.0 µg, average mass calculated from primary sequence Human, Recombinant 10.0 µg Histone H2B Human, Recombinant were is 13788.97 Da. This confirms the protein identity Note: The protein concentration (1 mg/ml, 73 µM) loaded on a 10–20% Tris-Glycine SDS-PAGE gel as well as the absence of any modifications of the is calculated using the molar extinction coefficient and stained with Coomassie Blue. The calculated histone. 1-800-632-7799 for Histone H2B (6400) and its absorbance at molecular weight is 13788.97 Da. Its apparent [email protected] 280 nm (3,4). 1.0 A units = 2.2 mg/ml N-terminal Protein Sequencing: Protein identity 280 molecular weight on 10–20% Tris-Glycine SDS- www.neb.com was confirmed using Edman Degradation to PAGE gel is ~17 kDa. M2505S 003140416041 Synonyms: Histone H2B/q, Histone H2B.1, sequence the intact protein. Histone H2B-GL105 Mass Spectrometry: The mass of purified Histone H2B Human, Recombinant is 13788.5 Da Protease Assay: After incubation of 10 µg of M2505S B r kDa 1 2 3 4 5 6 7 as determined by ESI-TOF MS (Electrospray Histone H2B Human, Recombinant with a standard 250 4.0 mixture of proteins for 2 hours at 37°C, no 100 µg 1.0 mg/ml Lot: 0031404 150 13788.5 100 proteolytic activity could be detected by SDS- RECOMBINANT Store at –20°C Exp: 4/16 80 PAGE. 60 Description: Histone H2B combines with Histone 50 3.0 ) 6 Exonuclease Assay: Incubation of a 50 µl H2A to form the H2A-H2B heterodimer. Two H2A/ 40 30 reaction containing 10 µg of Histone H2B Human, H2B heterodimers interact with an H3/H4 tetramer 25 Recombinant with 1 µg of a mixture of single and to form the histone octamer. (1,2) Histone H2B is 2.0 3 , Counts (x10 double-stranded [ H] E. coli DNA (200,000 cpm/ also modified by various enzymes and can act as 20 15 µg) for 4 hours at 37°C released < 0.1% of the total a substrate for them. These modifications have Intensity 10 radioactivity. been shown to be important in gene regulation. 1.0 SDS-PAGE analysis of Histone H2B Human, Recombinant. Endonuclease Assay: Incubation of a 50 µl Source: An E.coli strain that carries a plasmid Lane 1 and 7: NEB Protein Ladder (NEB #P7703), Lane 2-6: reaction containing 10 µg of Histone H2B Human, encoding the cloned human histone H2B gene, 0.5 µg, 1.0 µg, 2.0 µg, 5.0 µg, 10.0 µg Histone H2B Human, Recombinant. 0.0 Recombinant with 1 µg of φX174 RF I HIST2H2BE or H2BFQ. (Genbank accession 10000 12000 14000 16000 18000 Mass (Da) number: AY131979) (See other side) ESI-TOF Analysis of Histone H2B Human, Recombinant. CERTIFICATE OF ANALYSIS (suprecoiled) plasmid DNA for 4 hours at 37°C resulted in < 5.0% conversion to RF II form (nicked circle) as determined by agarose gel electrophoresis. Protein Sequence: PEPAKSAPAPKKGSKKAVTK AQKKDGKKRKRSRKESYSIYVYKVLKQVHPDTGI SSKAMGIMNSFVNDIFERIAGEASRLAHYNKRST ITSREIQTAVRLLLPGELAKHAVSEGTKAVTKYTSSK (Genbank accession number: AAN59961) References: 1. Kornberg, R.D. (1977) Annu. Rev. Biochem. 46, 931–954. 2. van Holde, K.E. (1989) Chromatin, 1–497. 3. Gill, S.C. and von Hippel, P.H. (1989) Anal. Biochem. 182, 319–326. 4. Pace, C.N. et al. (1995) Protein Science, 4, 2411–2423. Page 2 (M2505) (suprecoiled) plasmid DNA for 4 hours at 37°C resulted in < 5.0% conversion to RF II form (nicked circle) as determined by agarose gel electrophoresis. Protein Sequence: PEPAKSAPAPKKGSKKAVTK AQKKDGKKRKRSRKESYSIYVYKVLKQVHPDTGI SSKAMGIMNSFVNDIFERIAGEASRLAHYNKRST ITSREIQTAVRLLLPGELAKHAVSEGTKAVTKYTSSK (Genbank accession number: AAN59961) References: 1. Kornberg, R.D. (1977) Annu. Rev. Biochem. 46, 931–954. 2. van Holde, K.E. (1989) Chromatin, 1–497. 3. Gill, S.C. and von Hippel, P.H. (1989) Anal. Biochem. 182, 319–326. 4. Pace, C.N. et al. (1995) Protein Science, 4, 2411–2423. Page 2 (M2505).
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