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17-10498 ChIPAb+ Monomethyl-Histone H3 (Lys36) - ChIP Validated Antibody and Primer Set

The ChIPAb+ Monomethyl-Histone H3 (Lys36) set includes the monomethyl-Histone H3 (Lys36) antibody, a Normal Rabbit IgG & control primers which amplify a 213 bp region of the human GAPDH coding region.

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Key Applications: Western Blotting, Chromatin Immunoprecipitation (ChIP), Immunoprecipitation, Dot Blot Research Category: Epigenetics & Nuclear Function Research Sub-Category: Histones, Chromatin Biology
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17-10498
25 assays  25 assays per set. Recommended use: 3 μL (1 μg) of antibody per chromatin immunoprecipitation (dependent upon biological context).
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      Key Spec Table

      Species ReactivityKey Applications
      M, H, RWB, ChIP, IP, DB
      Description
      Catalogue Number17-10498
      Brand Family Upstate
      Trade Name
      • ChIPAb+
      • Upstate
      DescriptionChIPAb+ Monomethyl-Histone H3 (Lys36) - ChIP Validated Antibody and Primer Set
      OverviewAll 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+™ Monomethyl-Histone H3 (Lys36) set includes the monomethyl-Histone H3 (Lys36) antibody, a Normal Rabbit IgG, and control primers which amplify a 213 bp region of the human GAPDH coding region. The Monomethyl-Histone H3 (Lys36) and negative controls are supplied in a scalable "per ChIP" reaction size and can be used to functionally validate the precipitation of Monomethyl-Histone H3 (Lys36) associated chromatin.
      Alternate Names
      • H3 histone family, member T
      • Histone H3
      • Histone cluster 3
      • H3
      References
      Product Information
      FormatAffinity Purified
      Control
      • Includes normal rabbit IgG and primers specific for human GAPDH coding region
      PresentationAnti-Monomethyl-Histone H3 (Lys36) (Rabbit Polyclonal). One vial containing 75 µL 0.35 mg/mL purified rabbit polyclonal in buffer containing 0.1 M Tris-Glycine (pH7.4),150mM NaCl, and 0.05% sodium azide, before the addition of 30% glycerol. Store at -20°C.

      Normal Rabbit IgG. One vial containing 125 µg Rabbit IgG in 125 µL storage buffer containing 0.05% sodium azide. Store at -20°C.

      ChIP Primers, GAPDH coding D2. One vial containing 75 μL of 5 μM of each primer specific for human GAPDH coding region. Store at -20°C.
      FOR: GCC ATG TAG ACC CCT TGA AGA G
      REV: ACT GGT TGA GCA CAG GGT ACT TTA T
      Quality LevelMQ100
      Applications
      ApplicationThe ChIPAb+ Monomethyl-Histone H3 (Lys36) set includes the monomethyl-Histone H3 (Lys36) antibody, a Normal Rabbit IgG & control primers which amplify a 213 bp region of the human GAPDH coding region.
      Key Applications
      • Western Blotting
      • Chromatin Immunoprecipitation (ChIP)
      • Immunoprecipitation
      • Dot Blot
      Application NotesChromatin Immunoprecipitation:

      Sonicated chromatin prepared from HeLa cells (1e5 cell equivalents per IP) were subjected to chromatin immunoprecipitation using 1 µg of Normal Rabbit IgG, or 3 µL (1 µg) Anti-Monomethyl-Histone H3 (Lys36) and the Magna ChIP™ HiSens (Cat. # 17-10460). Successful immunoprecipitation of monomethyl-Histone H3 (Lys36) associated DNA fragments was verified by qPCR using ChIP Primers, GAPDH coding D2 as a positive locus, and GAPDH promoter as a negative locus.

      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 Magna ChIP™ HiSens (Cat. # 17-10460) or EZ-MagnaChIP™ HiSens(Cat. # 17-10461) protocol for experimental details.

      Western Blotting Analysis:
      HeLa acid extract and various recombinant Histone proteins were probed with Anti-monomethyl-Histone H3 (Lys36) (0.05 µg /mL).

      Immunoprecipitation Analysis:

      10 µg from a representative lot immunoprecipitated monomethyl-Histone H3 (Lys36) from HeLa acid extract.
      10 µg of this antibody immunoprecipitated monomethyl -Histone H3 (Lys36) from 500 µg of HeLa acid extract.
      Arrow indicates monomethyl-Histone H3 (Lys36) (~17 kDa).

      Dot Blot Specificity Analysis:

      Unmodified and various modified Histone peptides were probed with Anti-monomethyl-Histone H3 (Lys36) (0.5 µg/mL).Histone peptides with various modifications were transferred to PVDF membrane (Cat. #: 16-665) and probed with Anti-monomethyl-Histone H3 (Lys36) at (0.5 µg/mL). Proteins were visualized using a Donkey anti-rabbit IgG conjugated to HRP and a chemiluminescence detection system.
      Biological Information
      ImmunogenKLH-conjugated linear peptide corresponding to the N-terminus of human Histone H3 monomethylated at Lys36.
      EpitopeMonomethylated Histone H3 Lys36
      HostRabbit
      SpecificityThis antibody recognizes the N-terminus of Histone H3 monomethylated at Lys36.
      Species Reactivity
      • Mouse
      • Human
      • Rat
      Species Reactivity NoteDemonstrated to react with human, mouse and rat. Wide species cross reactivity is predicted based on sequence similarity.
      Antibody TypePolyclonal Antibody
      Entrez Gene Number
      Entrez Gene SummaryHistones 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. [provided by RefSeq]
      Gene Symbol
      • HIST3H3
      • H3FT
      • H3/t
      • H3t
      • H3/g
      • H3.4
      • H3T
      UniProt Number
      UniProt SummaryFunction: 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.

      Subunit structure: 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, Chromosome

      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.

      Post-translational modification: Methylation at Lys-36 (H3K36me) is linked to gene activation.

      Sequence similarities: 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 AssuranceChromatin Immunoprecipitation:
      Sonicated chromatin prepared from HeLa cells (1e5 cell equivalents per IP) were subjected to chromatin immunoprecipitation using 1 µg of either Normal Rabbit IgG, or 3 µL (1 µg) Anti-Monomethyl-Histone H3 (Lys36) and the Magna ChIP™ HiSens Kit (Cat. # 17-10460).
      Successful immunoprecipitation of monomethyl Histone H3 (Lys36) associated DNA fragments was verified by qPCR using ChIP Primers, GAPDH coding D2. Please refer to the MagnaChIP™ HiSens (Cat. # 17-10460) or EZ-MagnaChIP™ HiSens (Cat. # 17-10461) 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 ConditionsStable 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 Size25 assays
      Material Package25 assays per set. Recommended use: 3 μL (1 μg) of antibody per chromatin immunoprecipitation (dependent upon biological context).
      Transport Information
      Supplemental Information
      Specifications
      Global Trade Item Number
      Catalogue Number GTIN
      17-10498 04053252870408