17-10269 ChIPAb+™ Phospho-Histone H3 (Ser28) Antibody

17-10269
25 assays  25 assays per set. Recommended use: 5 μL of antibody per chromatin immunoprecipitation (dependent upon biological context).
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      Overview

      Replacement Information

      Key Specifications Table

      Species ReactivityKey Applications
      H, M WB, ICC, DB, PIA, ChIP
      Description
      Catalogue Number17-10269
      Trade Name
      • ChIPAb+
      • Upstate
      DescriptionChIPAb+™ Phospho-Histone H3 (Ser28) Antibody
      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+™ Phospho-Histone H3 (Ser28) set includes the Phospho-Histone H3 (Ser28) antibody, a Normal Rabbit IgG, and control primers which amplify a 166 bp region of the human GAPDH promoter. The Phospho-Histone H3 (Ser28) 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 (Ser28)-associated chromatin.
      Alternate Names
      • Histone H3.1t
      • H3/t
      • H3t
      • H3/g
      Background InformationHistone 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
      FormatAffinity Purified
      Control
      • Includes normal rabbit IgG and primers specific for human GAPDH promoter
      PresentationAnti- Phospho-Histone H3 (Ser28) (Rabbit Polyclonal). One vial containing 125 μl of purified rabbit polyclonal IgG in buffer containing 0.1 M Tris-Glycine (pH 7.4), 150 mM NaCl with 0.05% sodium azide before the addition of 30% glycerol. Store at -20°C.
      Normal Rabbit IgG. Two vials containing 75 μL of normal rabbit IgG in buffer containing 0.01M PBS with 0.1% sodium azide. Store at -20°C.
      ChIP Primers, human GAPDH promoter. One vial containing 75 μL of 5 μM 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
      Applications
      ApplicationThis ChIPAb+ Phospho-Histone H3 (Ser28) -ChIP Validated Antibody & Primer Set conveniently includes the antibody & the specific control PCR primers.
      Key Applications
      • Western Blotting
      • Immunocytochemistry
      • Dot Blot
      • Peptide Inhibition Assay
      • Chromatin Immunoprecipitation (ChIP)
      Application NotesChromatin Immunoprecipitation:
      Representative lot data.
      Sonicated chromatin prepared from untreated or colcemid-treated HeLa cells (1 X 106 cell equivalents per IP) were subjected to chromatin immunoprecipitation using either 5 µl Normal rabbit IgG, or 5 µl Anti-phospho-Histone H3 (Ser28) and the Magna ChIP™ A Kit (Cat. # 17-610). Successful immunoprecipitation of phospho-Histone H3 (Ser28)- associated DNA fragments was verified by qPCR using ChIP Primers, human GAPDH promoter region for untreated and treated chromatin samples (Figure 2). 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.
      HeLa acid extract (lane 1) and recombinant Histone H3 (lane 2) were probed with Anti-phospho-Histone H3 (Ser28) (1:2,000 dilution).
      Proteins were visualized using a Donkey Anti-Rabbit IgG secondary antibody conjugated to HRP and a chemiluminescence detection system. (Figure 3)

      Arrow indicates phospho-Histone H3 (Ser28) (~17 kDa).
      Dot Blot Specificity Analysis:
      Representative lot data.
      Histone peptides with various modifications (see table) were probed with Anti-phospho-Histone H3 (Ser28). Proteins were visualized using a Donkey Anti-Rabbit IgG secondary antibody conjugated to HRP and a chemiluminescence detection system. (Figure 4)
      Immunocytochemistry Analysis
      Representative lot data
      Confocal fluorescent analysis of HeLa cells using Anti-phospho-Histone H3 (Ser28) (Red). Actin filaments have been labeled with AlexaFluor® 488 dye-Phalloidin (Green). Nucleus is stained with DAPI (Blue). This antibody positively stains the mitotic cell nucleus. (Figure 5)
      Peptide Inhibition Assay: A 1:500 dilution from a representative lot was blocked by phospho-Histone H3 (Ser28) peptides, but not non-specific peptides.
      Biological Information
      ImmunogenKLH-conjugated linear peptide corresponding to Histone H3 phosphorylated at Ser28.
      EpitopePhosphorylated Ser28
      HostRabbit
      Species Reactivity
      • Human
      • Mouse
      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.
      Gene Symbol
      • HIST3H3
      • H3FT
      Modifications
      • Phosphorylation
      Purification MethodAffinity Purfied
      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.
      Molecular Weight~17 kDa observed
      Physicochemical Information
      Dimensions
      Materials Information
      Toxicological Information
      Safety Information according to GHS
      Safety Information
      Product Usage Statements
      Quality AssuranceChromatin Immunoprecipitation:
      Representative lot data.
      Sonicated chromatin prepared from colcemid-treated HeLa cells (1 X 106 cell equivalents per IP) were subjected to chromatin immunoprecipitation using either 5 µl Normal rabbit IgG, or 5 µl Anti-phospho-Histone H3 (Ser28) and the Magna ChIP™ A Kit (Cat. # 17-610). Successful immunoprecipitation of phospho-Histone H3 (Ser28)-associated DNA fragments was verified by qPCR using ChIP Primers, human GAPDH promoter region (Figure 1).
      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 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.
      Note: Variability in freezer temperatures below -20°C may cause glycerol containing solutions to become frozen during storage.
      Packaging Information
      Material Size25 assays
      Material Package25 assays per set. Recommended use: 5 μL of antibody per chromatin immunoprecipitation (dependent upon biological context).
      Transport Information
      Supplemental Information
      Specifications

      Documentation

      ChIPAb+™ Phospho-Histone H3 (Ser28) Antibody SDS

      Title

      Safety Data Sheet (SDS) 

      ChIPAb+™ Phospho-Histone H3 (Ser28) Antibody Certificates of Analysis

      TitleLot Number
      ChIPAb+ Phospho-Histone H3 (Ser28) - 17-1026917-10269
      ChIPAb+ Phospho-Histone H3 (Ser28) - Q2012440Q2012440
      ChIPAb+™ Phospho-Histone H3 (Ser28) - 29253132925313
      ChIPAb+™ Phospho-Histone H3 (Ser28) - 24615692461569

      References

      Reference overviewPub Med ID
      Interleukin-1beta and fibroblast growth factor receptor 1 cooperate to induce cyclooxygenase-2 during early mammary tumourigenesis.
      Reed, Johanna R, et al.
      Breast Cancer Res., 11: R21 (2009)  2009

      Show Abstract
      19393083 19393083
      Steel factor controls midline cell death of primordial germ cells and is essential for their normal proliferation and migration.
      Runyan, C; Schaible, K; Molyneaux, K; Wang, Z; Levin, L; Wylie, C
      Development (Cambridge, England)  133  4861-9  2006

      Show Abstract
      17107997 17107997
      Identification of a novel phosphorylation site on histone H3 coupled with mitotic chromosome condensation.
      Goto, H, et al.
      J. Biol. Chem., 274: 25543-9 (1999)  1999

      Show Abstract
      10464286 10464286
      Increased phosphorylation of histone H1 in mouse fibroblasts transformed with oncogenes or constitutively active mitogen-activated protein kinase kinase.
      Chadee, D N, et al.
      J. Biol. Chem., 270: 20098-105 (1995)  1995

      Show Abstract
      7650028 7650028

      Brochure

      Title
      Advance your Epigenetics Research (MilliporeSigma)