Mononucleosomes Recombinant Protein | HIST1H2AC recombinant protein

Recombinant Mononucleosomes (TH2B)-biotinylated

Cat. Num. AAA388260

• Gene Names: HIST1H2AC; H2A/l; H2AFL; dJ221C16.4
• Purity: The recombinant protein is >95% pure by SDS-PAGE.
• Synonyms: Mononucleosomes; Recombinant Mononucleosomes (TH2B)-biotinylated; TH2B; testis-specific H2B; testis-specific Histone 2B; HIST1H2AC recombinant protein

• Host: E Coli
• Purity/Purification: The recombinant protein is >95% pure by SDS-PAGE.
• Form/Format: Recombinant Mononucleosomes (TH2B)-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 (TH2B)-biotinylated, consist 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 NP_003503.1), TH2B that includes amino acids 1-127 (end)(accession number NP_733759.1), 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 (TH2B)-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 (TH2B) - biotinylated Mononucleosomes (TH2B) 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 (TH2B) are pulled down by streptavidin beads. * indicates streptavidin.)

Testing Data

(Recombinant Mononucleosomes (TH2B) - biotinylated DNA gel Biotinylated Mononucleosomes (TH2B) were run on a 2% agarose gel and stained with ethidium bromide. Lane 1: 601 DNA which was used for assembly of nucleosomes. Lane 2: MW. Lane 3: Intact 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 Recombinant Mononucleosomes (TH2B) - biotinylated 2 ug Recombinant Mononucleosomes (TH2B) were incubated with DOT1L in reaction buffer for 3 h at room temperature. Western Blot was used for detecting the generation of reaction products (H3K79me2). DOT1L only and mononucleosomes only were used as negative control. The Western Blot results show that mononucleosomes are a more suitable substrate for DOT1L than histone octamers. Nucleosomes = Mononucleosomes.)

Related Product Information for HIST1H2AC 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 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 biology processes including chromosome segregation, DNA repair, transcriptional regulation and mRNA processing. TH2B (testis-specific H2B) is originally identified as a testis-specific variant of histone H2B. TH2A is a testis-specific variant of histone H2A. They are abundant in the testis, oocytes and fertilized eggs. TH2A and TH2B are controlled by a shared promoter that is located between them. Disruption of Th2a and Th2b genes causes defects in spermatogenesis. TH2A/TH2B may enhance reprogramming by introducing processes that normally operate in zygotes and during SCNT. TH2A and TH2B induce nucleosome instability. TH2B controls the chromatin-to-nucleoprotamine 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 HIST1H2AC recombinant protein

Recombinant Proteins; Histone

NCBI and Uniprot Product Information

• NCBI GI #: 4504245
• NCBI GeneID: 8334
• NCBI Accession #: NP_003503.1; NP_733759.1; NP_003520.1; NP_003539.1
• NCBI GenBank Nucleotide #: NM_003512.3
• UniProt Accession #: Q93077; O00775; O00776; O00777; O00778; Q540R1; B2R4F7
• Molecular Weight: 14,105 Da
• NCBI Official Full Name: histone H2A type 1-C
• NCBI Official Synonym Full Names: histone cluster 1 H2A family member c
• NCBI Official Symbol: HIST1H2AC
• NCBI Official Synonym Symbols: H2A/l; H2AFL; dJ221C16.4
• NCBI Protein Information: histone H2A type 1-C
• UniProt Protein Name: Histone H2A type 1-C
• UniProt Gene Name: HIST1H2AC
• UniProt Synonym Gene Names: H2AFL

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 is intronless and encodes a replication-dependent histone that is a member of the histone H2A family. Transcripts from this gene lack polyA tails but instead contain a palindromic termination element. This gene is found in the large histone gene cluster on chromosome 6. [provided by RefSeq, Aug 2015]

Uniprot Description

Core component of nucleosome. 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.