SDS-Page
(Recombinant Mononucleosomes H3.3 (K18I) – biotin 12.5% SDS-PAGE with Coomassie blue staining MW: 108 kDa Purity: >92%)
Mononucleosomes H3.3 Recombinant Protein | H3.3 recombinant protein
Recombinant Mononucleosomes H3.3 (K18I)-biotin
Synonyms
Mononucleosomes H3.3; Recombinant Mononucleosomes H3.3 (K18I)-biotin; H3.3 recombinant protein
Host
E Coli
Form/Format
Recombinant Mononucleosomes H3.3 (K18I)-biotin are supplied in 10mM Tris-HCl pH8.0, 1mM EDTA, 2mM DTT, and 20% glycerol.
Protein Species
Human
Notes
Recombinant Mononucleosomes H3.3 (K18I)-biotin is suitable for use as substrates for histone modification enzymes, or to generate chromatin in vitro.
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
SDS-Page
(Recombinant Mononucleosomes H3.3 (K18I) – biotin 12.5% SDS-PAGE with Coomassie blue staining MW: 108 kDa Purity: >92%)
SDS-Page
(Recombinant Mononucleosomes H3.3 (K18I) – biotin 12.5% SDS-PAGE with Coomassie blue staining MW: 108 kDa Purity: >92%)
Testing Data
(Recombinant Mononucleosomes H3.3 (K18I) - biotin DNA gel Recombinant Mononucleosomes H3.3 (K18I) - biotin were run on a 2% agarose gel and stained with ethidium bromide. Lane 1: DNA marker. Lane 2: Free 601 DNA which was used for assembly of nucleosome. Lane 3: Intact mononucleosomes H3.3 (K18I) - biotin. Intact mononucleosomes H3.3 (K18I) - biotin migrated much higher than free 601 DNA. The agarose gel shows that almost all of 601 DNA wrapped histone octamers to form nucleosomes.)
Testing Data
(Recombinant Mononucleosomes H3.3 (K18I) - biotin DNA gel Recombinant Mononucleosomes H3.3 (K18I) - biotin were run on a 2% agarose gel and stained with ethidium bromide. Lane 1: DNA marker. Lane 2: Free 601 DNA which was used for assembly of nucleosome. Lane 3: Intact mononucleosomes H3.3 (K18I) - biotin. Intact mononucleosomes H3.3 (K18I) - biotin migrated much higher than free 601 DNA. The agarose gel shows that almost all of 601 DNA wrapped histone octamers to form nucleosomes.)
SDS-Page
(Streptavidin pull down assay for Recombinant Mononucleosomes H3.3 (K18I) – biotin 24 ug biotinylated mononucleosomes were incubated with 10 ul streptavidin beads for 1hr at 4 degree C. Streptavidin beads were washed 3 times with 1 ml binding buffer. Then the beads were added to 60 &mixeo;l 2×SDS loading buffer and boiled for 10 min at 95°&C. 2.4 micro;l samples were loaded and run on a 12.5% SDS-PAGE gel and stained by Commassie blue. The SDS-PAGE gel shows that almost all of biotinylated mononucleosomes were pulled down by streptavidin beads.)
SDS-Page
(Streptavidin pull down assay for Recombinant Mononucleosomes H3.3 (K18I) – biotin 24 ug biotinylated mononucleosomes were incubated with 10 ul streptavidin beads for 1hr at 4 degree C. Streptavidin beads were washed 3 times with 1 ml binding buffer. Then the beads were added to 60 &mixeo;l 2×SDS loading buffer and boiled for 10 min at 95°&C. 2.4 micro;l samples were loaded and run on a 12.5% SDS-PAGE gel and stained by Commassie blue. The SDS-PAGE gel shows that almost all of biotinylated mononucleosomes were pulled down by streptavidin beads.)
Related Product Information for H3.3 recombinant protein
Short Description: Recombinant Mononucleosomes H3.3 (K18I)-biotin consist of a 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) with a point mutation Lys18Ile, and H4 that includes amino acids 1-103 (end)(accession number NM_003548). All of these histones were expressed in E Coli cells. The molecular weight of the histone octamer is 108 kDa. Recombinant Mononucleosomes H3.3 (K18I)-biotin is suitable for use as substrates for histone modification enzymes, or to generate chromatin in vitro.
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 signifcantly more active, as well as specifc, 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. Histone H3.1 and Histone H3.3 are the two main Histone H3 variants found in plants and animals. They are known to be important for gene regulation. Histone H3.1 and H3.3 have been shown to demonstrate unique genomic localization patterns thought to be associated with their specific functions in regulation of gene activity. Specifically, Histone H3.3 primarily colocalizes with marks associated with gene activation (H3K4me3, H2BK120ub1, and RNA pol II occupancy). Deposition of the Histone H3.1 variant into the nucleosome correlates with the canonical DNA synthesis-dependent deposition pathway, whereas Histone H3.3 primarily serves as the replacement Histone H3 variant outside of S-phase, such as during gene transcription. Mutations in histone H3 occur with high genetic penetrance within rare paediatric gliomas and sarcomas. Researchers found that mutations at H3 N-terminal residues at or near PTM sites including R2, R8, K18 and R26 might act as oncohistones.
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 signifcantly more active, as well as specifc, 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. Histone H3.1 and Histone H3.3 are the two main Histone H3 variants found in plants and animals. They are known to be important for gene regulation. Histone H3.1 and H3.3 have been shown to demonstrate unique genomic localization patterns thought to be associated with their specific functions in regulation of gene activity. Specifically, Histone H3.3 primarily colocalizes with marks associated with gene activation (H3K4me3, H2BK120ub1, and RNA pol II occupancy). Deposition of the Histone H3.1 variant into the nucleosome correlates with the canonical DNA synthesis-dependent deposition pathway, whereas Histone H3.3 primarily serves as the replacement Histone H3 variant outside of S-phase, such as during gene transcription. Mutations in histone H3 occur with high genetic penetrance within rare paediatric gliomas and sarcomas. Researchers found that mutations at H3 N-terminal residues at or near PTM sites including R2, R8, K18 and R26 might act as oncohistones.
Product Categories/Family for H3.3 recombinant protein
NCBI and Uniprot Product Information
NCBI Accession #
NCBI GenBank Nucleotide #
Molecular Weight
Molecular weight of the histone octamer is 108kDa
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Product Notes
The H3.3 (Catalog #AAA389372) 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 H3.3, 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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