Product Class: Generic Enzyme

Histone H2B Human, Recombinant

Catalog #SizeConcentration
M2505S100 μg1 mg/ml

Description

Histone H2B combines with Histone H2A to form the H2A-H2B heterodimer. Two H2A/H2B heterodimers interact with an H3/H4 tetramer to form the histone octamer. Histone H2B is also modified by various enzymes and can act as a substrate for them. These modifications have been shown to be important in gene regulation.

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 H2B Human, Recombinant.
Lane 1 and 7: NEB Protein Ladder (NEB #P7703 ), Lane 2-6: 0.5, 1.0, 2.0, 5.0, 10.0 µg Histone H2B Human, Recombinant.
ESI-TOF Analysis of Histone H2B Human, Recombinant.

Properties and Usage

Storage Temperature

-20°C

Storage Conditions

300 mM NaCl
1 mM EDTA
20 mM sodium phosphate
pH 7.0 @ 25°C

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.
  • 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.
  • Molecular Weight Determination (Mass Spectrometry) :
    The molecular weight of the product is determined using mass spectrometry.
  • N-terminal Protein Sequencing :
    Protein identity is confirmed using Edman Degradation to sequence the N-terminus of the intact protein.
  • 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.

Supporting Documents

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.

Notes

  1. 1 mg/ml, 73 µM is calculated using the molar extinction coefficient for Histone H2B (6400) and its absorbance at 280 nm (3,4). 1.0 A280 units = 2.2 mg/ml

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. (1988). Anal. Biochem.. 182, 319-326.
  4. Pace, C.N. et al. (1995). Protein Science. 4, 2411-2423.
  1. Do the histones need to be reconstituted?
  2. What are the recommended histone storage conditions?
  3. Are the histones fusion proteins or tagged proteins?
  4. Can the histones be used as substrates for protein modification enzymes? Which ones?
After thawing on ice, mix well by pipetting the solution up and down. Do not centrifuge.