Mononucleosomes Recombinant Protein | H2AFX recombinant protein

Recombinant Mononucleosomes (H2A.X)-biotinylated

Cat. Num. AAA388284

• Gene Names: H2AFX; H2AX; H2A.X; H2A/X
• Synonyms: Mononucleosomes; Recombinant Mononucleosomes (H2A.X)-biotinylated; H2A Histone Family Member X; H2A/X; H2AFX recombinant protein

• Host: E Coli
• Form/Format: Recombinant Mononucleosomes (H2A.X)-biotinylated (20ug protein + 19.2ug DNA) are supplied in 10mM Tris-HCl, pH8.0, 1mM EDTA, 2mM DTT and 20% glycerol.

• Protein Species: Human
• Conjugation: Biotinylated; Tag
• Protein Details: Recombinant Mononucleosomes (H2A.X)-biotinylated, consist of 167 bp of 601 DNA with a 5' biotin tag and two molecules each of histones H2A.X that includes amino acids 1-143 (end)(accession number NP_002096.1) with a C-terminal 6xHis-tag, 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 cells.
• Notes: Recombinant Mononucleosomes (H2A.X)-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

(Recombinant Mononucleosomes (H2A.X) - biotinylated, SDS-PAGE. 12.5% SDS-PAGE gel stained with Coomassie Blue. Purity: ? 95%)

SDS-Page

(Streptavidin pull-down of Recombinant Mononucleosomes (H2A.X) - biotinylated. Mononucleosomes (H2A.X) were pulled down with 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 gel result shows that almost all biotinylated mononucleosomes (H2A.X) are pulled down by Streptavidin beads. *Indicates Streptavidin. Nucleosomes = Mononucleosomes)

Testing Data

(Recombinant Mononucleosomes (H2A.X) - biotinylated, DNA gel. Biotinylated mononucleosomes (H2A.X) were run on a 2% agarose gel and stained with ethidium bromide. Lane 1: DNA marker. Lane 2: free 601 DNA. Lane 3: Intact mononucleosomes.Intact mononucleosomes migrate much higher than free 601 DNA. The agarose gel result shows that almost all of 601 DNA wraps histone octamers to form mononucleosomes.)

Related Product Information for H2AFX 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 of four different types: H2A, H2B, H3 and H4. Histones are subject to posttranslational modifications, such as methylation, acetylation, phosphorylation, mono-ubiquitination, etc. 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 biology processes including chromosome segregation, DNA repair, transcriptional regulation and mRNA processing. H2A.X (also known as H2AFX, Histone Family Member X) is a histone H2A family member. In mammals, variant histone H2A.X represents 10% of the total H2A, whereas in yeast 90% of H2A is of the H2A.X class. The main function of H2AX is associated with the DNA damage repair (DDR) system which is induced by DNA double strand breaks (DSBs). In many species, including S. cerevisiae, Xenopus laevis, and Drosophila melanogaster, the fourth serine residue from the carboxyl-terminal end of histone H2A becomes phosphorylated. In mammals, this event is related to phosphorylation of the H2A.X variant histone on serine 139 by DNA-dependent protein kinases (ATR/ATM/DNA- PK). The phosphorylated form of H2A.X is commonly denoted as g-H2A.X. This H2A.XS139 phosphorylation facilitates the recruitment of the DNA damage repair machinery, as well as chromatin remodelers such as INO80 and SWR1. H2A.X is a regulator of the epithelial-mesenchymal transition. 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 H2AFX recombinant protein

Recombinant Proteins; Histone

NCBI and Uniprot Product Information

• NCBI GI #: 4504253
• NCBI GeneID: 3014
• NCBI Accession #: NP_002096.1; NP_003509.1; NP_003520.1; NP_003539.1
• NCBI GenBank Nucleotide #: NM_002105.2
• UniProt Accession #: P16104; Q4ZGJ7; Q6IAS5
• Molecular Weight: 15,145 Da
• NCBI Official Full Name: histone H2A.x
• NCBI Official Synonym Full Names: H2A histone family, member X
• NCBI Official Symbol: H2AFX
• NCBI Official Synonym Symbols: H2AX; H2A.X; H2A/X
• NCBI Protein Information: histone H2A.x; H2AX histone
• UniProt Protein Name: Histone H2A.x
• UniProt Gene Name: H2AFX
• UniProt Synonym Gene Names: H2AX; H2a/x
• UniProt Entry Name: H2AX_HUMAN

NCBI Description

Histones are basic nuclear proteins that are responsible for the nucleosome structure of the chromosomal fiber in eukaryotes. Two molecules of each of the four core histones (H2A, H2B, H3, and H4) form an octamer, around which approximately 146 bp of DNA is wrapped in repeating units, called nucleosomes. The linker histone, H1, interacts with linker DNA between nucleosomes and functions in the compaction of chromatin into higher order structures. This gene encodes a member of the histone H2A family, and generates two transcripts through the use of the conserved stem-loop termination motif, and the polyA addition motif. [provided by RefSeq, Jul 2008]

Uniprot Description

H2AX: a histone that replaces conventional H2A in a subset of nucleosomes. Required for checkpoint-mediated arrest of cell cycle progression in response to low doses of ionizing radiation and for efficient repair of DNA double strand breaks (DSBs) specifically when modified by C-terminal phosphorylation. Histones are basic nuclear proteins that are responsible for the nucleosome structure of the chromosomal fiber in eukaryotes. Two molecules of each of the four core histones (H2A, H2B, H3, and H4) form an octamer, around which approximately 146 bp of DNA is wrapped in repeating units, called nucleosomes. 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. DNA accessibility is regulated via a complex set of post-translational modifications of histones, also called histone code, and nucleosome remodeling. Phosphorylated on S139 by ATM and DNA-PK in response to DNA double strand breaks (DSBs) generated by exogenous genotoxic agents and by stalled replication forks, and may also occur during meiotic recombination events and immunoglobulin class switching in lymphocytes. Phosphorylation can extend up to several thousand nucleosomes from the actual site of the DSB and may mark the surrounding chromatin for recruitment of proteins required for DNA damage signaling and repair. Widespread phosphorylation may also serve to amplify the damage signal or aid repair of persistent lesions. Dephosphorylation of S139 by PP2A is required for DNA DSB repair. Apparently phosphorylated on Y143 by WSTF, determining the relative recruitment of either DNA repair or pro-apoptotic factors. H2AXpY142 favors the recruitment of pro-apoptotic factors APBB1 and JNK1. In contrast, dephosphorylation of pY143 by EYA phosphatases favors the recruitment of MDC1-containing DNA repair complexes to the tail of phosphorylated pS139. Monoubiquitination of K119 by RING1 and RNF2/RING2 complex gives a specific tag for epigenetic transcriptional repression. Following DNA double-strand breaks (DSBs), it is ubiquitinated through 'K63' linkages by the E2 ligase UBE2N and the E3 ligases RNF8 and RNF168, leading to the recruitment of repair proteins to sites of DNA damage.

Protein type: Helicase; DNA-binding; DNA repair, damage

Chromosomal Location of Human Ortholog: 11q23.3

Cellular Component: male germ cell nucleus; nucleoplasm; XY body; chromosome, telomeric region; condensed nuclear chromosome; nuclear chromatin; replication fork; nucleosome; nucleus

Molecular Function: protein binding; enzyme binding; DNA binding; histone binding; protein heterodimerization activity; damaged DNA binding

Biological Process: nucleosome assembly; positive regulation of DNA repair; meiotic cell cycle; double-strand break repair; DNA damage checkpoint; spermatogenesis; response to ionizing radiation; DNA repair; response to DNA damage stimulus; double-strand break repair via homologous recombination