Application Data
(Recombinant Polynucleosomes (H3.1) protein activity assay. Results show that polynucleosomes are better substrates than octamers for NSD1-SET and NSD2-SET. Lanes 5-9 represent polynucleosomes that have been subjected to subsequent freeze-thaw cycles to demonstrate their stability. Activity was detected by fluorography.)
Polynucleosomes Recombinant Protein
Recombinant Polynucleosomes (H3.1)
Purity
The recombinant protein is >95% pure by SDS-PAGE..
Synonyms
Polynucleosomes; N/A; Recombinant Polynucleosomes (H3.1); Polynucleosomes recombinant protein
Host
E Coli
Purity/Purification
The recombinant protein is >95% pure by SDS-PAGE..
Form/Format
Recombinant Polynucleosomes (H3.1)(20ug protein + 24ug DNA) are supplied at a concentration of 0.55ug/ul in 10mM Tris-HCl, pH8.0, 1mM EDTA, 2mM DTT and 20% glycerol.
Protein Species
Human
Protein Details
Recombinant Polynucleosomes (H3.1), Human consists of 5000 bp of DNA (plasmid pG5E4) and two molecules each of histones H2A that includes amino acids 1-130 (end)(accession number NM_003512), H2B that includes amino acids 1-126 (end)(accession number NM_003518), H3.1 that includes amino acids 1-136 (end)(accession number NM_003529), and H4 that includes amino acids 1-103 (end)(accession number NM_003548). Plasmid pG5E4 contains 9 of 5S rDNA nucleosome positioning sequences of L. variegatus, 5 of GAL4 binding sites and E4 promoter. Every 5S rDNA can wrap one histone octamer to form a nucleosome. 5 of GAL4 binding sites and E4 promoter can wrap histone octamers to form dinucleosomes. A pG5E4 plasmid can warp histone octamers to form 11 nucleosomes.
Notes
Recombinant Polynucleosomes (H3.1) are suitable for use in the study of enzyme kinetics, inhibitor screening, and selectivity profiling. HMT Assay Conditions: 50mM TrisCl, pH 8.6, 0.02% Triton X-100, 2mM MgCl2, 1mM TCEP, 100uM SAM, 30 ng/ul Recombinant Polynucleosomes (H3.1), 30 ng/ul DOT1L (1-416 aa) at 2 hours at room temperature. Activity was detected by fluorography.
Dry Ice Shipment
Extra charge fee may add to your shipping cost as dry ice is required to ship this product.
Preparation and Storage
Recombinant proteins in solution are temperature sensitive and must be stored at -80 degree C to prevent degradation. Avoid repeated freeze/thaw cycles and keep on ice when not in storage.
Shipping Temp: Dry Ice
Shipping Temp: Dry Ice
Application Data
(Recombinant Polynucleosomes (H3.1) protein activity assay. Results show that polynucleosomes are better substrates than octamers for NSD1-SET and NSD2-SET. Lanes 5-9 represent polynucleosomes that have been subjected to subsequent freeze-thaw cycles to demonstrate their stability. Activity was detected by fluorography.)
Application Data
(Recombinant Polynucleosomes (H3.1) protein activity assay. Results show that polynucleosomes are better substrates than octamers for NSD1-SET and NSD2-SET. Lanes 5-9 represent polynucleosomes that have been subjected to subsequent freeze-thaw cycles to demonstrate their stability. Activity was detected by fluorography.)
Application Data
(Recombinant Polynucleosomes (H3.1) protein DNA Gel-shift assay Polynucleosomes (H3.1) and free plasmid DNA were run on a 1% agarose gel and stained with ethidium bromide. Intact polynucleosomes migrate higher than free DNA, thus the DNA resolves at a higher molecular weight when nucleosome-bound. * represents the mixture of DNA and histone octamers (incorrect assembly).)
Application Data
(Recombinant Polynucleosomes (H3.1) protein DNA Gel-shift assay Polynucleosomes (H3.1) and free plasmid DNA were run on a 1% agarose gel and stained with ethidium bromide. Intact polynucleosomes migrate higher than free DNA, thus the DNA resolves at a higher molecular weight when nucleosome-bound. * represents the mixture of DNA and histone octamers (incorrect assembly).)
SDS-PAGE
(Recombinant Polynucleosomes (H3.1) protein gel. Recombinant Polynucleosomes (H3.1) run on an SDS-PAGE gel and stained with Coomassie Blue.)
SDS-PAGE
(Recombinant Polynucleosomes (H3.1) protein gel. Recombinant Polynucleosomes (H3.1) run on an SDS-PAGE gel and stained with Coomassie Blue.)
Related Product Information for Polynucleosomes recombinant protein
Short Description: Our Recombinant Polynucleosomes (H3.1) consists of 5000 bp of DNA (plasmid) and two molecules each of histones H2A that includes amino acids 1-130 (end)(accession number NM_003512), H2B that includes amino acids 1-126 (end)(accession number NM_003518), H3.1 that includes amino acids 1-136 (end)(accession number NM_003529), and H4 that includes amino acids 1-103 (end)(accession number NM_003548). Recombinant Polynucleosomes (H3.1) was generated in E Coli, and has an observed molecular weight of 108 kDa. Nucleosomes comprise the smallest subunit of chromatin; they consist of 147 bp of DNA wrapped around an octamer of core histone proteins (H2A, H2B, H3 and H4). Therefore, nucleosomes are more physiologically relevant substrates than histones and histone-derived peptides for use in in vitro studies. More importantly, some histone methyltransferases are significantly more active, as well as specific, when using nucleosomal substrates in HMT assays, such as DOT1L and NSD family enzymes.
Background: In vivo, histones are wrapped around by DNA in chromatin. Therefore, nucleosomes are more physiologically relevant substrates than histones and histone-derived peptides for in vitro studies. More importantly, some histone methyltransferases are significantly more active, as well as specific, when using nucleosomal substrates in HMT assays, such as DOT1L and NSD family enzymes. Nucleosomes are also widely used in histone methyltransferase screening assays to identify small molecular inhibitors for drug discovery.
Background: In vivo, histones are wrapped around by DNA in chromatin. Therefore, nucleosomes are more physiologically relevant substrates than histones and histone-derived peptides for in vitro studies. More importantly, some histone methyltransferases are significantly more active, as well as specific, when using nucleosomal substrates in HMT assays, such as DOT1L and NSD family enzymes. Nucleosomes are also widely used in histone methyltransferase screening assays to identify small molecular inhibitors for drug discovery.
Product Categories/Family for Polynucleosomes recombinant protein
NCBI and Uniprot Product Information
NCBI Accession #
NCBI GenBank Nucleotide #
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Product Notes
The Polynucleosomes (Catalog #AAA59700) is a Recombinant Protein produced from E Coli and is intended for research purposes only. The product is available for immediate purchase. It is sometimes possible for the material contained within the vial of "Polynucleosomes, Recombinant Protein" to become dispersed throughout the inside of the vial, particularly around the seal of said vial, during shipment and storage. We always suggest centrifuging these vials to consolidate all of the liquid away from the lid and to the bottom of the vial prior to opening. Please be advised that certain products may require dry ice for shipping and that, if this is the case, an additional dry ice fee may also be required.Precautions
All products in the AAA Biotech catalog are strictly for research-use only, and are absolutely not suitable for use in any sort of medical, therapeutic, prophylactic, in-vivo, or diagnostic capacity. By purchasing a product from AAA Biotech, you are explicitly certifying that said products will be properly tested and used in line with industry standard. AAA Biotech and its authorized distribution partners reserve the right to refuse to fulfill any order if we have any indication that a purchaser may be intending to use a product outside of our accepted criteria.Disclaimer
Though we do strive to guarantee the information represented in this datasheet, AAA Biotech cannot be held responsible for any oversights or imprecisions. AAA Biotech reserves the right to adjust any aspect of this datasheet at any time and without notice. It is the responsibility of the customer to inform AAA Biotech of any product performance issues observed or experienced within 30 days of receipt of said product. To see additional details on this or any of our other policies, please see our Terms & Conditions page.Item has been added to Shopping Cart
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