Mononucleosomes Recombinant Protein | H2AFB2 recombinant protein

Recombinant Mononucleosomes (H2A.Bbd)-biotinylated

Cat. Num. AAA388259

• Gene Names: H2AFB2; H2A.Bbd
• Purity: The recombinant protein is >95% pure by SDS-PAGE.
• Synonyms: Mononucleosomes; Recombinant Mononucleosomes (H2A.Bbd)-biotinylated; H2A.Bbd; H2AFB2 recombinant protein

• Host: E Coli
• Purity/Purification: The recombinant protein is >95% pure by SDS-PAGE.
• Form/Format: Recombinant Mononucleosomes (H2A.Bbd)-biotinylated (20ug protein + 20ug DNA) are supplied in 10mM Tris-HCl, pH8.0, 1mM EDTA, 2mM DTT and 20% glycerol. Please refer to product insert upon arrival for lot-specific concentration.

• Protein Species: Human
• Conjugation: Biotinylated
• Protein Details: Recombinant Mononucleosomes (H2A.Bbd)-biotinylated, consist of 167 bp of 601 DNA with a 5' biotin tag and two molecules each of histones H2A.Bbd that includes amino acids 1-115 (end)(accession number NP_001017991.1), H2B that includes amino acids 1-126 (end)(accession number NP_003509.1), H3 that includes amino acids 1-136 (end)(accession number NP_003520.1), and H4 that includes amino acids 1-103 (end)(accession number NP_003539.1). All of these histones were expressed in E Coli.
• Notes: Recombinant Mononucleosomes (H2A.Bbd)-biotinylated, are suitable for use as substrates in the study of enzyme kinetics, inhibitor screening, and selectivity profiling.
• 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

SDS-Page

(Streptavidin Pull-down for Recombinant Mononucleosomes (H2A.Bbd) - biotinylated Mononucleosomes (H2A.Bbd) were pulled down by streptavidin beads. Input mononucleosomes (Lane 1) and the mononucleosomes pulled down by streptavidin (Lane 2) were run on a 12.5% SDS-PAGE gel and stained with Coomassie Blue.The SDS-PAGE gel result shows that more than 80% biotinylated nucleosomes (H2A.Bbd) are pulled down by streptavidin beads. * indicates streptavidin.)

Testing Data

(Recombinant Mononucleosomes (H2A.Bbd) - biotinylated DNA gel Biotinylated Mononucleosomes (H2A.Bbd) were run on a 2% agarose gel and stained with ethidium bromide. Lane 1: Intact mononucleosomes. Lane 2: 601 DNA which was used for assembly of mononucleosomes. Intact mononucleosomes migrate much higher than free DNA. The agarose gel result shows that almost all of 601 DNA wrap histone octamers to form mononucleosomes.)

Western Blot (WB)

(Western Blot analysis for Mononucleosomes (H2A.Bbd) - biotinylated 2 ug Recombinant Mononucleosomes (H2A.Bbd) were incubated with DOT1L in reaction buffer for 3 h at room temperature. Western Blot was used for detecting the generation of reaction products (H3K79me1). DOT1L only and mononnucleosomes only were used as negative control.The Western Blot result shows that mononucleosomes are more suitable substrate for DOT1L than histone octamers.)

Related Product Information for H2AFB2 recombinant protein

Short Description: 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. Nucleosomes are also widely used in histone methyltransferase screening assays to identify small molecular inhibitors for drug discovery.

Background: In vivo, the nucleosome is the basic structural unit of chromatin. It is comprised of about 146 bp of DNA wrapped around a core of eight histones, made up of of four different types: H2A, H2B, H3 and H4. Histones are subject to posttranslational modifications, such as methylation, acetylation, phosphorylation, and mono-ubiquitination. Histone modifications influence multiple chromatin templated processes such as gene transcription, DNA repair and recombination. Besides the "major" histones, there are some histone variants in specific regions of chromatin or in specific cell types. Histone variants are involved in multiple processes, including chromosome segregation, DNA repair, transcriptional regulation and mRNA processing. H2A.Bbd (Barr Body Deficient) is a histone H2A family member. H2A.Bbd is one of the most rapidly evolving variants, which shares 48% sequence identity with the major H2A histone. H2A.Bbd is excluded from the inactive X chromosome and is enriched in active regions of genome. H2A.Bbd is enriched at sites of DNA synthesis. H2A.Bbd nucleosomes are less stable than those bearing the major H2A histone and to organize only 118 bp of DNA, indicating that DNA in H2A.Bbd chromatin may be more accessible and, thus, more amenable to transcription. H2A.Bbd is involved in mammalian spermiogenesis. 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 H2AFB2 recombinant protein

Recombinant Proteins; Histone

NCBI and Uniprot Product Information

• NCBI GI #: 63029943
• NCBI GeneID: 474381
• NCBI Accession #: NP_001017991.1; NP_003509.1; NP_003520.1; NP_003539.1
• NCBI GenBank Nucleotide #: NM_001017991.2
• UniProt Accession #: P0C5Z0; P98176; Q5TZB2; Q6FG78; Q96PR7; A1L4E4
• Molecular Weight: 12,713 Da
• NCBI Official Full Name: histone H2A-Bbd type 2/3
• NCBI Official Synonym Full Names: H2A histone family member B2
• NCBI Official Symbol: H2AFB2
• NCBI Official Synonym Symbols: H2A.Bbd
• NCBI Protein Information: histone H2A-Bbd type 2/3
• UniProt Protein Name: Histone H2A-Bbd type 2/3
• Protein Family: Histone
• UniProt Gene Name: H2AFB2
• UniProt Synonym Gene Names: H2ABBD; H2AFB; H2A.Bbd

NCBI Description

Histones are basic nuclear proteins that are responsible for the nucleosome structure of the chromosomal fiber in eukaryotes. Nucleosomes consist of approximately 146 bp of DNA wrapped around a histone octamer composed of pairs of each of the four core histones (H2A, H2B, H3, and H4). The chromatin fiber is further compacted through the interaction of a linker histone, H1, with the DNA between the nucleosomes to form higher order chromatin structures. This gene encodes a replication-independent histone that is a member of the histone H2A family. This gene is part of a region that is repeated three times on chromosome X, once in intron 22 of the F8 gene and twice closer to the Xq telomere. This record represents the middle copy. [provided by RefSeq, Oct 2015]

Uniprot Description

Atypical histone H2A which can replace conventional H2A in some nucleosomes and is associated with active transcription and mRNA processing. Nucleosomes wrap and compact DNA into chromatin, limiting DNA accessibility to the cellular machineries which require DNA as a template. Histones thereby play a central role in transcription regulation, DNA repair, DNA replication and chromosomal stability. Nucleosomes containing this histone are less rigid and organize less DNA than canonical nucleosomes in vivo. They are enriched in actively transcribed genes and associate with the elongating form of RNA polymerase. They associate with spliceosome components and are required for mRNA splicing. May participate in spermatogenesis.