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Histone H2A Human, Recombinant |
 | | | | Histone H2A Human, Recombinant | | | |
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Prices are in US dollars and valid only for US orders.
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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. (1,2) Histone H2A 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:
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)
Figure 1: SDS-PAGE analysis of Histone H2A Human, Recombinant. Lane 1: NEB Protein Marker, Broad Range (NEB #P7702), Lane 2: 2 µg Histone H2A Human, Recombinant, Lane 3: 4 µg Histone H2A Human, Recombinant. (please see Quality Control section below for more information.)
Figure 2: ESI-TOF Analysis of Histone H2A Human, Recombinant. (Please see Quality Control section below for more information.)
Source:
An E.coli strain that carries a plasmid encoding the cloned human Histone H2A gene, HIST3H2A. (Genbank accession number: AY131974)
Properties & Usage
Molecular Weight:
Theoretical: 13,990.28 daltons
Reaction & Storage Conditions
Concentration:
1 mg/ml
Storage Conditions:
20 mM sodium phosphate
300 mM NaCl
1 mM DTT
1 mM EDTA
pH 7.0 @ 25°C
Storage Temperature:
-20°C
Notes
General notes:- The protein concentration (1 mg/ml or 71 µM) is calculated using the molar extinction coefficient for Histone H2A (3840) and its absorbance at 280 nm (3,4). 1.0 A280 units = 3.6 mg/ml
FAQs
- Do the histones need to be reconstituted?
- What are the recommended histone storage conditions?
- Are the histones fusion proteins or tagged proteins?
- Can the histones be used as substrates for protein modification enzymes? Which ones?
Quality Control for Current Lot
Quality control values for a specific lot can be found on the datacard which accompanies each product.
SDS-PAGE:
2 µg and 4 µg of Histone H2A Human, Recombinant were loaded on a 10-20% Tris-Tricine SDS-PAGE gel and stained with Coomassie Blue. The calculated molecular weight is 13990.28 Da. Its apparent molecular weight on 10-20% Tris-Tricine SDS-PAGE gel is ~14 kDa. (see Figure 1)
Mass Spectrometry:
The mass of purified Histone H2A Human, Recombinant is 13990 Da as determined by ESI-TOF MS (Electrospray Ionization-Time of Flight Mass Spectrometry). The average mass calculated from primary sequence is 13990.28 Da. This confirms the protein identity as well as the absence of any modifications of the histone. (see Figure 2)
N-terminal Protein Sequencing:
Protein identity was confirmed using Edman Degradation to sequence the intact protein.
Protease Assay:
After incubation of 10 µg of Histone H2A Human, Recombinant with a standard mixture of proteins for 2 hours at 37°C, no proteolytic activity could be detected by SDS-PAGE.
Exonuclease Assay:
Incubation of a 50 µl reaction containing 10 µg of Histone H2A Human, Recombinant with 1 µg of a mixture of single and double-stranded [3H] E. coli DNA (200,000 cpm/µg) for 4 hours at 37°C released < 0.1% of the total radioactivity.
Endonuclease Assay:
Incubation of a 50 µl reaction containing 10 µg of Histone H2A Human, Recombinant with 1 µg of ΦX174 RF I (supercoiled) 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.
References
- Kornberg, R.D. (1977) Annu. Rev. Biochem., 46, 931-954.
- van Holde, K.E. (1989) Chromatin, 1-497.
- Gill, S.C. and von Hippel, P.H. (1989) Anal. Biochem., 182, 319-326.
- Pace, C.N. et al. (1995) Protein Science, 4, 2411-2423.
Companion Products
Histone H10 Human, Recombinant
Histone H2B Human, Recombinant
Histone H3.1 Human, Recombinant
Histone H4 Human, Recombinant
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