Histone H2A/H2B Dimer Human, Recombinant

Description

Histone H2A combines with Histone H2B to form the H2A/H2B heterodimer. Two H2A/H2B heterodimers interact with an H3/H4 tetramer to form the histone octamer. Histones are also modified by various enzymes and can act as substrates for them. These modifications have been shown to be important in gene regulation. Because the histones are folded with their subunit partners, the dimer may be a better substrate for specific enzymes and modifications.

Protein Sequence:
Human Histone H2A Protein Sequence: SGRGK QGGKA RAKAK SRSSR AGLQF PVGRV HRLLR KGNYS ERVGA GAPVY LAAVL EYLTA EILEL AGNAA RDNKK TRIIP RHLQL AIRND EELNK LLGRV TIAQG GVLPN IQAVL LPKKT ESHHK AKGK (Genbank accession number: AAN59960)

Human Histone H2B Protein Sequence: PEPAK SAPAP KKGSK KAVTK AQKKD GKKRK RSRKE SYSIY VYKVL KQVHP DTGIS SKAMG IMNSF VNDIF ERIAG EASRL AHYNK RSTIT SREIQ TAVRL LLPGE LAKHA VSEGT KAVTK YTSSK(Genbank accession number: AAN59961)

SDS-PAGE analysis of Histone H2A/H2B Dimer, Recombinant
SDS-PAGE analysis of Histone H2A/H2B Dimer, Recombinant
Lane 1&7: NEB Protein Ladder (NEB #P7703)
Lane 2: Histone H2A (NEB #M2502S)
Lane 3: Histone H2B (NEB #M2505S)
Lane 4–6: 2.0, 4.0, 8.0 µg Histone H2A/H2B Dimer

Product Source

Purified H2A and H2B (NEB #M2502 and NEB #M2505) were denatured, refolded and the dimer was purified by gel filtration.

Properties and Usage

Storage Temperature

-20°C

Storage Conditions

20 mM Tris-HCl
2 M NaCl
1 mM DTT
1 mM EDTA
pH 8.0 @ 25°C

Notes

  1. 20 μM, 0.55 mg/ml is calculated using the molar extinction coefficient for Histone H2A/H2B Dimer (11,920) and its absorbance at 280 nm (4, 5)
  2. To use as a substrate in an enzyme modification assay or other salt sensitive protocol, use 1 to 2 µl of the dimer in a minimum 20 µl reaction so that the salt concentration in the reaction ≤ 200 mM.
  3. Because of high salt, the solution does not always remain frozen.

References

  1. Kornberg, R.D. (1977). Annu. Rev. Biochem. 46, 931-954.
  2. Van Holde, K.E. (1989). Chromatin. 1-479.
  3. Mersha, F. unpublished observations.
  4. Gill, S.C. and von Hippel, P.H. (1989). Anal. Biochem. 182, 319-326.
  5. Pace, C.N. et al. (1995). Protein Science. 4, 2411-2423.

Tech Tips

To use as a substrate in an enzyme modification assay or other salt sensitive protocol, use 1 to 2 μl of the dimer in a minimum 20 μl reaction so that the salt concentration in the reaction = 200 mM.

Datacards

The Product Summary Sheet, or Data Card, includes details for how to use the product, as well as details of its formulation and quality controls. The following file naming structure is used to name the majority of these document files: [Catalog Number]Datasheet-Lot[Lot Number]. For those product lots not listed below, please contact NEB at info@neb.com or fill out the Technical Support Form for appropriate document.

Interactive Tools

Quality Control

Quality Control Assays

The following Quality Control Tests are performed on each new lot and meet the specifications designated for the product. Individual lot data can be found on the Product Summary Sheet/Datacard or Manual which can be found in the Supporting Documents section of this page. Further information regarding NEB product quality can be found here.
  • Endonuclease Activity (Nicking):
    The product is tested in a reaction containing a supercoiled DNA substrate. After incubation for 4 hours the percent converted to the nicked form is determined by agarose gel electrophoresis.
  • Exonuclease Activity (Radioactivity Release):
    The product is tested in a reaction containing a radiolabeled mixture of single and double-stranded DNA. After incubation for 4 hours the exonuclease activity is determined by the % release of radioactive nucleotides.
  • Protease Activity (SDS-PAGE):
    The product is tested for protease activity by incubation with a standard mixture of proteins resulting in no detectable degradation of the proteins as determined by SDS-PAGE.

Safety Data Sheet

The following is a list of Safety Data Sheet (SDS) that apply to this product to help you use it safely.

Datacards

The Product Summary Sheet, or Data Card, includes details for how to use the product, as well as details of its formulation and quality controls. The following file naming structure is used to name the majority of these document files: [Catalog Number]Datasheet-Lot[Lot Number]. For those product lots not listed below, please contact NEB at info@neb.com or fill out the Technical Support Form for appropriate document.