Supplied in: 20 mM Sodium Phosphate (pH 7.0), Quality Control Assays: Mass Spectrometry: The mass of purified Histone Histone H2A 300 mM NaCl and 1 mM EDTA. SDS-PAGE: 0.5, 1.0, 2.0, 5.0, 10.0 µg of Histone H2A Human, Recombinant is 13,990.72 Da as determined by ESI-TOF MS (Electrospray Human, Recombinant H2A Human, Recombinant were loaded on a Note: The protein concentration (1 mg/ml or 71 µM) 10–20% Tris-Glycine SDS-PAGE gel and stained Ionization-Time of Flight Mass Spectrometry). The is calculated using the molar extinction coefficient with Coomassie Blue. The calculated molecular average mass calculated from primary sequence 1-800-632-7799 for Histone H2A (3840) and its absorbance at weight is 13990.24 Da. Its apparent molecular is 13,990.24 Da. This confirms the protein identity [email protected] 280 nm (3,4). 1.0 A units = 3.6 mg/ml of the histone. www.neb.com 280 weight on 10–20% Tris-Glycine SDS-PAGE gel is ~15 kDa. kDa 1 2 3 4 5 6 7 Protease Assay: After incubation of 5 µg of His- 250 tone H2A Human, Recombinant with a standard 150 mixture of proteins for 4 hours at 37°C, no proteo- M2502S 100 80 lytic activity could be detected by SDS-PAGE. ) 13,990.72 100 µg 1.0 mg/ml Lot: 0151508 60 6 50 2 Exonuclease Assay: Incubation of a 50 µl RECOMBINANT Store at –20°C Exp: 8/17 40 reaction containing 10 µg of Histone H2A Human, 1

30 Counts (x 10 Recombinant with 1 µg of a mixture of single and Description: Histone H2A combines with Histone 14,033.82 3 H2B to form the H2A-H2B heterodimer. Two H2A/ 25 double-stranded [ H] E. coli DNA (200,000 cpm/ 0 µg) for 4 hours at 37°C released < 0.1% of the total H2B heterodimers interact with an H3/H4 tetramer 20 10,000 12,000 14,000 16,000 18,000 20,000 to form the histone octamer (1,2). Histone H2A is radioactivity. 15 Deconvoluted Mass (Da) also modified by various enzymes and can act as a 10 Endonuclease Assay: Incubation of a 50 µl substrate for them. These modifications have been ESI-TOF Analysis of Histone H2A Human, Recombinant. reaction containing 10 µg of Histone H2A shown to be important in gene regulation. SDS-PAGE analysis of Histone H2A Human, Recombinant. Lane 1&7: NEB Protein Ladder (NEB #P7703), Lane 2 thru 6: Human, Recombinant with 1 µg of φX174 RF I Source: An strain that carries a plasmid 0.5–10.0 µg Histone H2A Human, Recombinant. (Please see (supercoiled) plasmid DNA for 4 hours at 37°C E. coli Quality Control section for more information) encoding the cloned human Histone H2A resulted in < 5.0% conversion to RF II form gene, HIST3H2A. (Genbank accession number: (nicked circle) as determined by agarose gel electrophoresis. AY131974) (see other side) CERTIFICATE OF ANALYSIS

Supplied in: 20 mM Sodium Phosphate (pH 7.0), Quality Control Assays: Mass Spectrometry: The mass of purified Histone Histone H2A 300 mM NaCl and 1 mM EDTA. SDS-PAGE: 0.5, 1.0, 2.0, 5.0, 10.0 µg of Histone H2A Human, Recombinant is 13,990.72 Da as determined by ESI-TOF MS (Electrospray Human, Recombinant H2A Human, Recombinant were loaded on a Note: The protein concentration (1 mg/ml or 71 µM) 10–20% Tris-Glycine SDS-PAGE gel and stained Ionization-Time of Flight Mass Spectrometry). The is calculated using the molar extinction coefficient with Coomassie Blue. The calculated molecular average mass calculated from primary sequence 1-800-632-7799 for Histone H2A (3840) and its absorbance at weight is 13990.24 Da. Its apparent molecular is 13,990.24 Da. This confirms the protein identity [email protected] 280 nm (3,4). 1.0 A units = 3.6 mg/ml of the histone. www.neb.com 280 weight on 10–20% Tris-Glycine SDS-PAGE gel is ~15 kDa. kDa 1 2 3 4 5 6 7 Protease Assay: After incubation of 5 µg of His- 250 tone H2A Human, Recombinant with a standard 150 mixture of proteins for 4 hours at 37°C, no proteo- M2502S 100 80 lytic activity could be detected by SDS-PAGE. ) 13,990.72 100 µg 1.0 mg/ml Lot: 0151508 60 6 50 2 Exonuclease Assay: Incubation of a 50 µl RECOMBINANT Store at –20°C Exp: 8/17 40 reaction containing 10 µg of Histone H2A Human, 1

30 Counts (x 10 Recombinant with 1 µg of a mixture of single and Description: Histone H2A combines with Histone 14,033.82 3 H2B to form the H2A-H2B heterodimer. Two H2A/ 25 double-stranded [ H] E. coli DNA (200,000 cpm/ 0 µg) for 4 hours at 37°C released < 0.1% of the total H2B heterodimers interact with an H3/H4 tetramer 20 10,000 12,000 14,000 16,000 18,000 20,000 to form the histone octamer (1,2). Histone H2A is radioactivity. 15 Deconvoluted Mass (Da) also modified by various enzymes and can act as a 10 Endonuclease Assay: Incubation of a 50 µl substrate for them. These modifications have been ESI-TOF Analysis of Histone H2A Human, Recombinant. reaction containing 10 µg of Histone H2A shown to be important in gene regulation. SDS-PAGE analysis of Histone H2A Human, Recombinant. Lane 1&7: NEB Protein Ladder (NEB #P7703), Lane 2 thru 6: Human, Recombinant with 1 µg of φX174 RF I Source: An strain that carries a plasmid 0.5–10.0 µg Histone H2A Human, Recombinant. (Please see (supercoiled) plasmid DNA for 4 hours at 37°C E. coli Quality Control section for more information) encoding the cloned human Histone H2A resulted in < 5.0% conversion to RF II form gene, HIST3H2A. (Genbank accession number: (nicked circle) as determined by agarose gel electrophoresis. AY131974) (see other side) CERTIFICATE OF ANALYSIS Protein Sequence: SGRGKQGGKARAKAKSRSSR AGLQFPVGRVHRLLRKGNYSERVGAGAPVYLAAV LEYLTAEILELAGNAARDNKKTRIIPRHLQLAIRND EELNKLLGRVTIAQGGVLPNIQAVLLPKKTESHHKA KGK (Genbank accession number: AAN59960)

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. Bio- chem. 182, 319–326. 4. Pace, C.N. et al. (1995) Protein Science 4, 2411–2423.

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NEW ENGLAND BIOLABS® is a registered trademark of New England Biolabs, Inc. This product is intended for research purposes only. This product is not intended to be used for therapeutic or diagnostic purposes in humans or animals.

Page 2 (M2502)

Protein Sequence: SGRGKQGGKARAKAKSRSSR AGLQFPVGRVHRLLRKGNYSERVGAGAPVYLAAV LEYLTAEILELAGNAARDNKKTRIIPRHLQLAIRND EELNKLLGRVTIAQGGVLPNIQAVLLPKKTESHHKA KGK (Genbank accession number: AAN59960)

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. Bio- chem. 182, 319–326. 4. Pace, C.N. et al. (1995) Protein Science 4, 2411–2423.

ISO 9001 ISO 14001 ISO 13485 Registered Registered Registered Quality Environmental Medical Devices Management Management

NEW ENGLAND BIOLABS® is a registered trademark of New England Biolabs, Inc. This product is intended for research purposes only. This product is not intended to be used for therapeutic or diagnostic purposes in humans or animals.

Page 2 (M2502)