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48-602MAG
Buffer Detection Kit for Magnetic Beads
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17-10141
Sigma-AldrichChIPAb+™ Phospho-Histone H3 (Thr3) - ChIP Validated Antibody and Primer Set, rabbit monoclonal
This ChIPAb+ Phospho-Histone H3 (Thr3) -ChIP Validated Antibody & Primer Set conveniently includes the antibody & the specific control PCR primers.
More>>This ChIPAb+ Phospho-Histone H3 (Thr3) -ChIP Validated Antibody & Primer Set conveniently includes the antibody & the specific control PCR primers. Less<<
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ChIPAb+™ Phospho-Histone H3 (Thr3) - ChIP Validated Antibody and Primer Set, rabbit monoclonal
Overview
All ChIPAb+ antibodies are individually validated for chromatin precipitation, every lot, every time. Each ChIPAb+ antibody set includes control primers (tested every lot by qPCR) to biologically validate your IP results in a locus-specific context. The qPCR protocol and primer sequences are provided, allowing researchers to validate ChIP protocols when using our antibody in their chromatin context. Each set also includes a negative control antibody to ensure specificity of the ChIP reaction. The ChIPAb+ Phospho-Histone H3 (Thr3) set includes the Phospho-Histone H3 (Thr3) antibody, a negative control supernatant, and control primers which amplify a 166 bp region of human GAPDH promoter. The Phospho-Histone H3 (Thr3) and negative controls are supplied in a scalable "per ChIP" reaction size and can be used to functionally validate the precipitation of Phospho-Histone H3 (Thr3)-associated chromatin.
Alternate Names
H3T3P
Histone H3 (phospho T3)
Background Information
Histone H3 is one of the five main histone proteins involved in the structure of chromatin in eukaryotic cells. Featuring a main globular domain and a long N-terminal tail, H3 is involved with the structure of the nucleosomes of the 'beads on a string' structure.
The N-terminal tail of histone H3 protrudes from the globular nucleosome core and can undergo several different types of epigenetic modifications that influence cellular processes. These modifications include the covalent attachment of methyl or acetyl groups to lysine and arginine amino acids and the phosphorylation of serine or threonine.
References
Product Information
Format
Culture Supernatant
HS Code
3002 15 90
Control
Includes negative control supernatant and primers specific for human GAPDH promoter.
Presentation
Anti-Phospho-Histone H3 (Thr3) (rabbit monoclonal). One vial containing 50 µL of cultured supernantant in 0.05% sodium azide. Store at -20°C. Negative Control Supernatant. One vial containing 100 µL of cultured supernatant in 0.05% sodium azide. Store at -20°C. Control Primers, human GAPDH promoter. One vial containing 75 μL of 5 μM of each primer specific for human GAPDH. Store at -20°C. FOR: TAC TAG CGG TTT TAC GGG CG REV: TCG AAC AGG AGG AGC AGA GAG CGA
This ChIPAb+ Phospho-Histone H3 (Thr3) -ChIP Validated Antibody & Primer Set conveniently includes the antibody & the specific control PCR primers.
Key Applications
Western Blotting
Dot Blot
Chromatin Immunoprecipitation (ChIP)
Application Notes
Chromatin Immunoprecipitation: Representative lot data. Sonicated chromatin prepared from untreated or colcemid-treated HeLa cells (1 X 10E6 cell equivalents per IP) were subjected to chromatin immunoprecipitation using 2 µL of either Negative Control Supernatant, or 2 µL ofAnti-Phospho-Histone H3 (Thr3) and the Magna ChIP™ A Kit (Cat. # 17-610). Successful immunoprecipitation of Phospho-Histone H3 (Thr3) associated DNA fragments was verified by qPCR using ChIP Primers, human GAPDH promoter Region for untreated and treated chromatin samples. Data is presented as percent input of each IP sample relative to input chromatin for each amplicon and ChIP sample as indicated. Please refer to the EZ-Magna ChIP™ A (Cat. # 17-408) or EZ-ChIP™ (Cat. # 17-371) protocol for experimental details. Western Blot Analysis: Representative lot data. Recombinant histone H3 (Lane 1) and acid extracts from untreated (Lane 2) or colcemid treated (Lane 3) HeLa cells were resolved by electrophoresis, transferred to nitrocellulose and probed with anti-Phospho-Histone H3 (Thr3), clone JY325 (10 ng/mL). Proteins were visualized using a goat anti-rabbit secondary antibody conjugated to HRP and a chemiluminescence detection system. Arrow indicates Phospho- Histone H3 (~17 kDa). Dot Blot: Representative lot data. A dilution series of Histone H3 peptides containing the following modifications was made: Column 1: Phospho-threonine 3 Column 2: unmodified, containing Thr3 Column 3: Phospho-threonine 6 Column 4: Phospho-threonine 11 Column 5: Phospho-threonine 22 Column 6: Phospho-serine 10 Column 7: Phospho-threonine 80 Column 8: Phospho-serine 28 2 μL of each dilution was spotted onto PVDF membranes and probed with Anti-Phospho- Histone H3 (Thr3), clone JY325 (20 ng/mL) Peptides were visualized using a Goat Anti-Rabbit secondary antibody conjugated to HRP and a chemiluminescence detection system..
Biological Information
Immunogen
Peptide containing the sequence [RTtrimKQ] in which lysine 4 is trimethylated on human histone H3
Epitope
Phosphorylated Thr3
Clone
JY325
Host
Rabbit
Specificity
Histone H3 phosphorylated at Threonine 3; cross-reactivity with phospho-histone H3 Thr22 detected in some applications
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 is intronless and encodes a member of the histone H3 family. Transcripts from this gene lack polyA tails; instead, they contain a palindromic termination element. This gene is located separately from the other H3 genes that are in the histone gene cluster on chromosome 6p22-p21.3.
FUNCTION: SwissProt: Q16695 # 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. SIZE: 136 amino acids; 15508 Da SUBUNIT: The nucleosome is a histone octamer containing two molecules each of H2A, H2B, H3 and H4 assembled in one H3-H4 heterotetramer and two H2A-H2B heterodimers. The octamer wraps approximately 147 bp of DNA. SUBCELLULAR LOCATION: Nucleus. TISSUE SPECIFICITY: Expressed in testicular cells. DEVELOPMENTAL STAGE: Expressed during S phase, then expression strongly decreases as cell division slows down during the process of differentiation. PTM: Acetylation is generally linked to gene activation. Acetylation on Lys-10 impairs methylation at Arg-9. Acetylation on Lys-19 and Lys-24 favors methylation at Arg-18 (By similarity). & Citrullination at Arg-9 and/or Arg-18 by PADI4 impairs methylation and represses transcription (By similarity). & Asymmetric dimethylation at Arg-18 by CARM1 is linked to gene activation. Symmetric dimethylation at Arg-9 by PRMT5 is linked to gene repression (By similarity). & Methylation at Lys-5, Lys-37 and Lys-80 are linked to gene activation. Methylation at Lys-5 facilitates subsequent acetylation of H3 and H4. Methylation at Lys-80 is associated with DNA double-strand break (DSB) responses and is a specific target for TP53BP1. Methylation at Lys-10 and Lys-28 are linked to gene repression. Methylation at Lys-10 is a specific target for HP1 proteins (CBX1, CBX3 and CBX5) and prevents subsequent phosphorylation at Ser-11 and acetylation of H3 and H4. Methylation at Lys-5 and Lys-80 require preliminary monoubiquitination of H2B at 'Lys-120'. Methylation at Lys-10 and Lys-28 are enriched in inactive X chromosome chromatin (By similarity). & Phosphorylated at Thr-4 by GSG2/haspin during prophase and dephosphorylated during anaphase. At centromeres, specifically phosphorylated at Thr-12 from prophase to early anaphase. Phosphorylated at Ser-11 during the whole mitosis. Phosphorylation at Ser-11, which is linked to gene activation, prevents methylation at Lys-10 but facilitates acetylation of H3 and H4. Phosphorylated at Ser-29 by MLTK isoform 1, RPS6KA5 or AURKB during mitosis or upon ultraviolet B irradiation (By similarity). & Phosphorylation at 'Ser-11' is crucial for chromosome condensation and cell-cycle progression during mitosis and meiosis. In addition phosphorylation at 'Ser-11' is important during interphase because it enables the transcription of genes following external stimulation, like stress or growth factors. Phosphorylation at 'Ser-11' is also an essential regulatory mechanism for neoplastic cell transformation. Phosphorylation at 'Ser-11' by AURKB/Aurora-B mediates the dissociation of HP1 proteins (CBX1, CBX3 and CBX5) from heterochromatin. & Ubiquitinated (By similarity). SIMILARITY: SwissProt: Q16695 ## Belongs to the histone H3 family.
Molecular Weight
17 kDa
Physicochemical Information
Dimensions
Materials Information
Toxicological Information
Safety Information according to GHS
Safety Information
Product Usage Statements
Quality Assurance
Chromatin Immunoprecipitation: Representative lot data. Sonicated chromatin prepared from colcemid-treated HeLa cells (1 X 10E6 cell equivalents per IP) were subjected to chromatin immunoprecipitation using 2 µL of either Negative Control Supernatant, or 2 µL Anti-Phospho-Histone H3 (Thr3) and the Magna ChIP™ A Kit (Cat. # 17-610). Successful immunoprecipitation of Phospho-Histone H3 (Thr3) associated DNA fragments was verified by qPCR using ChIP Primers, human GAPDH promoter Region. Please refer to the EZ-Magna ChIP™ A (Cat. # 17-408) or EZ-ChIP™ (Cat. # 17-371) protocol for experimental details.
Usage Statement
Unless otherwise stated in our catalog or other company documentation accompanying the product(s), our products are intended for research use only and are not to be used for any other purpose, which includes but is not limited to, unauthorized commercial uses, in vitro diagnostic uses, ex vivo or in vivo therapeutic uses or any type of consumption or application to humans or animals.
Storage and Shipping Information
Storage Conditions
Stable for 1 year at -20°C from date of receipt. Handling Recommendations: Upon first thaw, and prior to removing the cap, centrifuge the vial and gently mix the solution. Aliquot into microcentrifuge tubes and store at -20°C. Avoid repeated freeze/thaw cycles, which may damage IgG and affect product performance.
Packaging Information
Material Size
25 assays
Material Package
25 assays per set. Recommended use: ~2 μL of antibody per chromatin immunoprecipitation (dependent upon biological context).
Transport Information
Supplemental Information
Specifications
Global Trade Item Number
Bestellnummer
GTIN
17-10141
04053252415913
Documentation
ChIPAb+™ Phospho-Histone H3 (Thr3) - ChIP Validated Antibody and Primer Set, rabbit monoclonal SDB
Millipore’s Histone H3 antibodies demonstrate specificity against histone H3. See below for acetyl-, methyl-, phospho- histone H3 Antibodies and Proteins, based on the expertise of Upstate & Chemicon. Weitere Informationen >>
