Supplied in: 20 mM Tris-HCl, pH 8.0 @ 25°C, 2 M Concentration: 20 µM (0.55 mg/ml) calculated Endonuclease Assay: Incubation of a 50 µl reaction H2A/H2B NaCl, 1 mM EDTA, 1 mM DTT. Store at –20°C. using the molar extinction coefficient of Histone Dimer containing 10 µg (~360 pmol) of Histone H2A/H2B Dimer Human, (Because of high salt, the solution does not always (11,920) and its absorbance at 280 nm (4,5). Dimer Human, Recombinant with 1 µg of φX174 remain frozen.) RF I (supercoiled) DNA for 4 hours at 37°C Recombinant Quality Control Assays: resulted in < 5.0% conversion to RF II form (nicked kDa 1 2 3 4 5 6 7 circle) as determined by agarose gel electrophoresis. 1-800-632-7799 [email protected] 250 SDS-PAGE: 2.0, 4.0, 8.0 µg of Histone H2A/H2B www.neb.com 150 Dimer Human, Recombinant were loaded on a References: M2508S 001120813081 100 80 10–20% Tris-Glycine SDS-PAGE gel and stained with 1. Kornberg, R.D. (1977) Annu. Rev. Biochem. 46, 60 Coomassie Blue. The calculated molecular weights 931–954. 50 of the two subunits found in this protein complex 2. van Holde, K.E. (1989) 1–497. 40 M2508S 30 are 13,990.28 Da for Histone H2A and 13,788.97 Da 3. Mersha, F., unpublished observations for . Their apparent molecular weight on 25 4. Gill, S.C. and von Hippel, P.H. (1989) Anal. Bio- 10–20% Tris-Glycine SDS-PAGE gel are ~14 kDa and 2 nmol 20 µM Lot: 0011208 chem. 182, 319–326. 20 ~14.5 kDa. RECOMBINANT Store at –20°C Exp: 8/13 15 5. Pace, C.N. et al. (1995) Protein Science 4, 10 Protease Assay: After incubation of 10 µg (~360 pmol) 2411–2423. Description: Histone H2A combines with Histone of Histone H2A/H2B Dimer Human, Recombinant with H2B to form the H2A/H2B heterodimer. Two H2A/H2B SDS-PAGE analysis of Histone H2A/H2B Dimer, Recombinant. heterodimers interact with an H3/H4 tetramer to form Lane 1&7: NEB Protein Ladder (NEB #P7703) a standard mixture of for 4 hours at 37°C, no the (1,2). are also modified Lane 2: Histone H2A (NEB #M2502S) proteolytic activity could be detected by SDS-PAGE. by various enzymes and can act as substrates for Lane 3: Histone H2B (NEB #M2505S) Lane 4–6: 2, 4, 8 µg Histone H2A/H2B Dimer Exonuclease Assay: Incubation of a 50 µl reaction them. These modifications have been shown to be containing 10 µg (~360 pmol) of .1/H4 important in gene regulation. Because the histones Tetramer Human, Recombinant with 1 µg of a mixture are folded with their subunit partners, the dimer Note: To use as a substrate in an enzyme modification of single and double-stranded [3H] E. coli DNA may be a better substrate for specific enzymes and assay or other salt sensitive protocol, use 1 to 2 µl of (200,000 cpm/µg) for 4 hours at 37°C released < 0.1% modifications (3). the dimer in a minimum 20 µl reaction so that the salt concentration in the reaction ≤ 200 mM. of the total radioactivity. Source: Purified H2A and H2B (NEB #M2502 and NWB #M2505) were denatured, refolded and purified CERTIFICATE OF ANALYSIS by gel filtration.

Supplied in: 20 mM Tris-HCl, pH 8.0 @ 25°C, 2 M Protein Concentration: 20 µM (0.55 mg/ml) calculated Endonuclease Assay: Incubation of a 50 µl reaction Histone H2A/H2B NaCl, 1 mM EDTA, 1 mM DTT. Store at –20°C. using the molar extinction coefficient of Histone Dimer containing 10 µg (~360 pmol) of Histone H2A/H2B Dimer Human, (Because of high salt, the solution does not always (11,920) and its absorbance at 280 nm (4,5). Dimer Human, Recombinant with 1 µg of φX174 remain frozen.) RF I (supercoiled) plasmid DNA for 4 hours at 37°C Recombinant Quality Control Assays: resulted in < 5.0% conversion to RF II form (nicked kDa 1 2 3 4 5 6 7 1-800-632-7799 circle) as determined by agarose gel electrophoresis. [email protected] 250 SDS-PAGE: 2.0, 4.0, 8.0 µg of Histone H2A/H2B www.neb.com 150 Dimer Human, Recombinant were loaded on a References: M2508S 001120813081 100 80 10–20% Tris-Glycine SDS-PAGE gel and stained with 1. Kornberg, R.D. (1977) Annu. Rev. Biochem. 46, 60 Coomassie Blue. The calculated molecular weights 931–954. 50 of the two subunits found in this protein complex 2. van Holde, K.E. (1989) Chromatin 1–497. 40 M2508S 30 are 13,990.28 Da for Histone H2A and 13,788.97 Da 3. Mersha, F., unpublished observations for Histone H2B. Their apparent molecular weight on 25 4. Gill, S.C. and von Hippel, P.H. (1989) Anal. Bio- 2 nmol 20 µM Lot: 0011208 10–20% Tris-Glycine SDS-PAGE gel are ~14 kDa and chem. 182, 319–326. 20 ~14.5 kDa. RECOMBINANT Store at –20°C Exp: 8/13 15 5. Pace, C.N. et al. (1995) Protein Science 4, 10 Protease Assay: After incubation of 10 µg (~360 pmol) 2411–2423. Description: Histone H2A combines with Histone of Histone H2A/H2B Dimer Human, Recombinant with H2B to form the H2A/H2B heterodimer. Two H2A/H2B SDS-PAGE analysis of Histone H2A/H2B Dimer, Recombinant. heterodimers interact with an H3/H4 tetramer to form Lane 1&7: NEB Protein Ladder (NEB #P7703) a standard mixture of proteins for 4 hours at 37°C, no the histone octamer (1,2). Histones are also modified Lane 2: Histone H2A (NEB #M2502S) proteolytic activity could be detected by SDS-PAGE. by various enzymes and can act as substrates for Lane 3: Histone H2B (NEB #M2505S) Lane 4–6: 2, 4, 8 µg Histone H2A/H2B Dimer Exonuclease Assay: Incubation of a 50 µl reaction them. These modifications have been shown to be containing 10 µg (~360 pmol) of Histone H3.1/H4 important in gene regulation. Because the histones Tetramer Human, Recombinant with 1 µg of a mixture are folded with their subunit partners, the dimer Note: To use as a substrate in an enzyme modification of single and double-stranded [3H] E. coli DNA may be a better substrate for specific enzymes and assay or other salt sensitive protocol, use 1 to 2 µl of (200,000 cpm/µg) for 4 hours at 37°C released < 0.1% modifications (3). the dimer in a minimum 20 µl reaction so that the salt concentration in the reaction ≤ 200 mM. of the total radioactivity. Source: Purified H2A and H2B (NEB #M2502 and NWB #M2505) were denatured, refolded and purified CERTIFICATE OF ANALYSIS by gel filtration.