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Histone H10 Human, Recombinant |
 | | | | Histone H10 Human, Recombinant | | | |
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Prices are in US dollars and valid only for US orders.
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Description:
Histone H1 acts on the linker region of polynucleosome DNA to condense the chromatin into structures of ~30 nm (1). It is not necessary for octamer or nucleosome core particle formation. Eight different Histone H1 proteins have been identified in the human genome (2). Histone H10 is a non replication-dependent histone that is highly expressed in cells that have terminally differentiated (3).
Protein Sequence:
TENST SAPAA KPKRA KASKK STDHP KYSDM IVAAI QAEKN RAGSS RQSIQ KYISH YKVGE NADSQ IKLSI KRLVT TGVLK QTKGV GASGS FRLAK SDEPK KSVAF KKTKK EIKKV ATPKK ASKPK KAASK APTKK PKATP VKKAK KKLAA TPKKA KKPKT VKAKP VKASK PKKAK PVKPK AKSSA KRAGK KK (Genbank accession number: P07305)
Figure 1: SDS-PAGE analysis of Histone H1° Human, Recombinant. Lane 1: NEB Protein Marker, Broad Range (NEB #P7702), Lanes 2 thru 6: 0.5, 1.0, 2.0, 5.0, 10.0 µg of Histone H1° Human, Recombinant. (Please see Quality Control section below for more information.)
Figure 2: ESI-TOF Analysis of Histone H1° Human, Recombinant. (Please see Quality Control section below for more information.)
Source:
An E.coli strain that carries a plasmid encoding the human histone H1 gene, H1F0 or H1FV. (Genbank accession number: X03473)
Properties & Usage
Molecular Weight:
Theoretical: 20,731.53 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, 48 µM) is calculated using the molar extinction coefficient for Histone H1 (3840) and its absorbance at 280 nm (4,5). 1.0 A280 units = 5.4 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:
0.5, 1.0, 2.0, 5.0, 10.0 µg of Histone H10 Human, Recombinant were loaded on a 10-20% Tris-Glycine SDS-PAGE gel and stained with Coomassie Blue. The calculated molecular weight is 20731.53 Da. Its apparent molecular weight on 10-20% Tris-Glycine SDS-PAGE gel is ~30 kDa.
Mass Spectrometry:
The mass of purified Histone H10 Human, Recombinant is 20732.26 Da as determined by ESI-TOF MS (Electrospray Ionization-Time of Flight Mass Spectrometry). The average mass calculated from primary sequence is 20731.53 Da. This confirms the protein identity as well as the absence of any modifications of the histone.
N-terminal Protein Sequencing:
Protein identity was confirmed using Edman Degradation to sequence the intact protein.
Enzyme Modification:
After incubation of a 30 µl reaction containing 50 units (pmol/min) of CDK2-cyclin A (NEB #P6025) for 1 minute at 30°C, 96 pmols of phosphate were transferred to Histone H10 Human, Recombinant (10 µM).
Protease Assay:
After incubation of 5 µg of Histone H10 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 H10 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 H10 Human, Recombinant with 1 µg of ΦX174 RF I (suprecoiled) 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
- van Holde, K.E. (1989) Chromatin, 1-497.
- Marzluff, W.F., et al. (2002) Genomics, 80, 487-497.
- Pehrson, J.R. and Cole, R.D. (1982) Biochem, 21, 456-460.
- 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 H2A Human, Recombinant
Histone H2B Human, Recombinant
Histone H3.1 Human, Recombinant
Histone H4 Human, Recombinant
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