WB (Western Blot)
(Recombinant Mononucleosomes (H3.3) - biotinylated activity. 2 ?g Mononucleosomes were incubated with 0 ?g, 0.25 ?g, 0.5 ?g, 1 ?g NSD2-SET in reaction buffer for 3 h at room temperature, respectively. Western blot was used for detecting the generation of reaction products (H3K36me2). Recombinant Histone Octamers was used as a control substrate. The Western blot result shows that mononucleosomes are a more suitable substrate for NSD2-SET than histone octamers.)
Mononucleosomes Recombinant Protein
Recombinant Mononucleosomes (H3.3)-biotinylated
Purity
The recombinant protein is >95% pure by SDS-PAGE.
Synonyms
Mononucleosomes; N/A; Recombinant Mononucleosomes (H3.3)-biotinylated; Mononucleosomes recombinant protein
Host
E Coli
Purity/Purification
The recombinant protein is >95% pure by SDS-PAGE.
Form/Format
Recombinant Mononucleosomes (H3.3)-biotinylated (20ug protein + 20ug DNA) is supplied in 10mM Tris-HCl, pH8.0, 1mM EDTA, 2mM DTT and 20% glycerol.
Protein Species
Human
Conjugation
Biotinylated
Protein Details
Recombinant Mononucleosomes (H3.3)-biotinylated consists of 167 bp of 601 DNA with 5' biotin tag 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), H3.3 that includes amino acids 1-136 (end)(accession number NM_005324), and H4 that includes amino acids 1-103 (end)(accession number NM_003548). All of these histones were expressed in E Coli cells.
Notes
Recombinant Mononucleosomes (H3.3)-biotinylated are suitable for use in the study of enzyme kinetics, inhibitor screening, and selectivity profiling. HMT Assay Conditions: 2ug mononucleosomes was incubated with different concentration of NSD2-SET in reaction buffer including 50mM TrisCl, pH 8.6, 0.02% Triton X-100, 2mM MgCl2, 1mM TCEP, 50uM SAM for 3 hours at room temperature. Activity was detected by Western blot.
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
WB (Western Blot)
(Recombinant Mononucleosomes (H3.3) - biotinylated activity. 2 ?g Mononucleosomes were incubated with 0 ?g, 0.25 ?g, 0.5 ?g, 1 ?g NSD2-SET in reaction buffer for 3 h at room temperature, respectively. Western blot was used for detecting the generation of reaction products (H3K36me2). Recombinant Histone Octamers was used as a control substrate. The Western blot result shows that mononucleosomes are a more suitable substrate for NSD2-SET than histone octamers.)
WB (Western Blot)
(Recombinant Mononucleosomes (H3.3) - biotinylated activity. 2 ?g Mononucleosomes were incubated with 0 ?g, 0.25 ?g, 0.5 ?g, 1 ?g NSD2-SET in reaction buffer for 3 h at room temperature, respectively. Western blot was used for detecting the generation of reaction products (H3K36me2). Recombinant Histone Octamers was used as a control substrate. The Western blot result shows that mononucleosomes are a more suitable substrate for NSD2-SET than histone octamers.)
Application Data
(Recombinant Mononucleosomes (H3.3) - biotinylated DNA gel. Mononucleosomes were run on an agarose gel and stained with ethidium bromide. Lane 1: 601 DNA which was used for assembly of mononucleosomes. Lane 2: Intact mononucleosomes. Intact mononucleosomes migrate much higher than free DNA. The agarose gel result showed almost all of the 601 DNA wrapped histone octamers to form mononucleosomes.)
Application Data
(Recombinant Mononucleosomes (H3.3) - biotinylated DNA gel. Mononucleosomes were run on an agarose gel and stained with ethidium bromide. Lane 1: 601 DNA which was used for assembly of mononucleosomes. Lane 2: Intact mononucleosomes. Intact mononucleosomes migrate much higher than free DNA. The agarose gel result showed almost all of the 601 DNA wrapped histone octamers to form mononucleosomes.)
SDS-PAGE
(Recombinant Mononucleosomes (H3.3) - biotinylated protein gel. 40 ug mononucleosomes were incubated with 15 ul streptavidin beads for 1 h at 4C. Samples were centrifuged at 4C, and supernatant was labeled as free histone octamers. Samples were run on a 12.5% SDS-PAGE gel and stained by Commassie blue. The SDS-PAGE gel result showed that more than 80% biotinylated mononucleosomes were pulled down by streptavidin beads. * indicates streptavidin.)
SDS-PAGE
(Recombinant Mononucleosomes (H3.3) - biotinylated protein gel. 40 ug mononucleosomes were incubated with 15 ul streptavidin beads for 1 h at 4C. Samples were centrifuged at 4C, and supernatant was labeled as free histone octamers. Samples were run on a 12.5% SDS-PAGE gel and stained by Commassie blue. The SDS-PAGE gel result showed that more than 80% biotinylated mononucleosomes were pulled down by streptavidin beads. * indicates streptavidin.)
Related Product Information for Mononucleosomes recombinant protein
Short Description: Our Recombinant Mononucleosomes (H3.3)-biotinylated 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), H3.3 that includes amino acids 1-136 (end)(accession number NM_005324) with a C-terminal Biotin tag, and H4 that includes amino acids 1-103 (end)(accession number NM_003548). Recombinant Mononucleosomes (H3.3)-biotinylated was generated in E Coli and has an observed molecular weight of 110 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. Recombinant Mononucleosomes (H3.3)-biotinylated are for enzyme kinetics and inhibitor screening.
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 Mononucleosomes recombinant protein
NCBI and Uniprot Product Information
NCBI Accession #
NCBI GenBank Nucleotide #
Molecular Weight
Molecular weight of histone octamer is 108kDa
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Product Notes
The Mononucleosomes (Catalog #AAA59703) 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 "Mononucleosomes, 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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