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17-671 ChIPAb+ Trimethyl-Histone H4 (Lys20) - ChIP Validated Antibody and Primer Set, rabbit monoclonal

This ChIPAb+ Trimethyl-Histone H4 (Lys20) -ChIP Validated Antibody & Primer Set conveniently includes the antibody & the specific control PCR primers.

MSDS (material safety data sheet) or SDS, CoA and CoQ, dossiers, brochures and other available documents.
Key Applications: Western Blotting, Chromatin Immunoprecipitation (ChIP) Research Category: Epigenetics & Nuclear Function Research Sub-Category: Chromatin Biology, Histones
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17-671
10 assays  10 assays per set. Recommended use: ~10 μL antibody per chromatin immunoprecipitation (dependent upon biological context).
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Species ReactivityKey Applications
H, Pl, VrtWB, ChIP
Description
Catalogue Number17-671
Brand Family Upstate
Trade Name
  • ChIPAb+
  • Upstate
DescriptionChIPAb+ Trimethyl-Histone H4 (Lys20) - ChIP Validated Antibody and Primer Set, rabbit monoclonal
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+ Trimethyl-Histone H4 (Lys20) set includes the Anti-trimethyl-Histone H4 (Lys20) antibody, a negative control supernatant, and qPCR primers which amplify a 110 bp region within the promoter of the humanβ-globin gene. The trimethyl histone H4 (Lys20) and negative control antibodies are supplied in a scalable "per ChIP" reaction size and can be used to functionally validate the precipitation of trimethyl-histone H4 (Lys20) associated chromatin.
Alternate Names
  • H4K20me3
  • Histone H4 (tri methyl K20)
  • H4 histone family, member A
  • histone 1
  • H4a
  • histone cluster 1
  • H4a
Product Information
FormatCulture Supernatant
Control
  • Includes negative control rabbit antibody and control primers specific for human β-globin promoter.
PresentationAnti-trimethyl-Histone H4 (Lys20) (rabbit monoclonal IgG). One vial containing 100 μL of cell culture supernatant in 0.1% sodium azide. Store at -20°C.

Negative Control Supernatant. One vial containing 100 μL cell rabbit monoclonal culture supernatant containing 0.1% sodium azide. Store at -20°C.

ChIP Primers B-Globin. One vial containing 75 μL of 5 μM of each primer specific for human β-globin. Store at -20°C.
FOR: AGG ACA GGT ACG GCT GTC ATC
REV: TTT ATG CCC AGC CCT GGC TC
Quality LevelMQ100
Applications
ApplicationThis ChIPAb+ Trimethyl-Histone H4 (Lys20) -ChIP Validated Antibody & Primer Set conveniently includes the antibody & the specific control PCR primers.
Key Applications
  • Western Blotting
  • Chromatin Immunoprecipitation (ChIP)
Application NotesChromatin Immunoprecipitation:
Sonicated chromatin prepared from HeLa cells (2 X 106 cell equivalents per IP) was subjected to chromatin immunoprecipitation using 10 μL of either a negative control supernatant or Anti-Trimethyl-Histone H4 (Lys20) antibody and the Magna ChIP A Kit (Part No. 17-610). Successful immunoprecipitation of trimethylhistone H4 (Lys20) associated DNA fragments was verified by qPCR using β-globin ChIP Primers versus Control Primers corresponding to the GAPDH promoter (Please see figures). Data is presented as percent input of each IP sample relative to input chromatin, with immunoprecipitated DNA from negative control supernatant shown as (-) and trimethyl histone H4 (Lys20) shown as (+).
Please refer to the EZ-Magna ChIP™ A (Cat. # 17-408) or EZ-ChIP™ (Cat. # 17-371) protocol for experimental details.

Western Blot Analysis:
Acid extracts from HeLa cells untreated or pre-absorbed with 1 μM of trimethylated (Lys20) Histone H4 peptide (Lanes 1 and 2 respectively) were probed with anti-trimethyl-Histone H4 (Lys20) (1:1,000) (Please see figures).

Dot Blot Analysis: Absurance Histone H3 Antibody Specificity Array (Cat. No. 16-667) and Absurance Histone H2A, H2B, H4 Antibody Specificity Array (Cat. No. 16-665), which contain histone peptides with various modifications were probed with Cat. No 04-079, Anti-trimethyl-Histone H4 (Lys20) ; dilution 1:500. Proteins were visualized using a Donkey anti-rabbit IgG conjugated to HRP and a chemiluminescence detection system.
Biological Information
ImmunogenThe trimethyl histone H4 (Lys20) antibody is made against a synthetic peptide corresponding to amino acids 18-22 of histone H4. trimethylated on lysine 20.
Epitopea.a. 18-22
HostRabbit
SpecificityRecognizes histone H4, Mr 11 kDa
IsotypeIgG
Species Reactivity
  • Human
  • Green Plants
  • Vertebrates
Species Reactivity NoteHuman. Not tested in other species. The immunogen sequence is identical in a wide range of animal and plant species, so broad cross-reactivity is expected.
Antibody TypeMonoclonal Antibody
Entrez Gene Number
Entrez Gene SummaryHistones 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 member of the histone H4 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.
Gene Symbol
  • H4/A
  • H4/B
  • H4/C
  • H4/D
  • H4/E
  • H4/G
  • H4/H
  • H4/I
  • H4/J
  • H4/J
  • H4/M
  • H4/N
  • H4/O
  • H4/a
  • H4F2
  • H4FA
  • H4FB
  • H4FC
  • H4FD
  • H4FE
  • H4FG
  • H4FH
  • H4FI
  • H4FJ
  • H4FK
  • H4FM
  • H4FN
  • H4FO
  • HIST2H4
Modifications
  • Methylation
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.

PTM: Acetylation at Lys-6, Lys-9, Lys-13 and Lys-17 occurs in coding regions of the genome but not in heterochromatin.

Citrullination at Arg-4 by PADI4 impairs methylation.

Monomethylation at Arg-4 by PRMT1 favors acetylation at Lys-9 and Lys-13. Demethylation is performed by JMJD6.

Monomethylated, dimethylated or trimethylated at Lys-21. Monomethylation is performed by SET8. Trimethylation is performed by SUV420H1 and SUV420H2 and induces gene silencing.

Ubiquitinated by the CUL4-DDB-RBX1 complex in response to ultraviolet irradiation. This may weaken the interaction between histones and DNA and facilitate DNA accessibility to repair proteins.

Sumoylated, which is associated with transcriptional repression.

SEQUENCE SIMILARITY: Belongs to the histone H4 family.

SEQUENCE CAUTION:The sequence AAI28106.1 differs from that shown. Reason: Frameshift at position 3.

Molecular Weight~11 kDa
Product Usage Statements
Quality AssuranceChromatin Immunoprecipitation:
Sonicated chromatin prepared from HeLa cells (1 X 106 cell equivalents per IP) were subjected to chromatin immunoprecipitation using 10 µL of either a negative control supernatant, or Anti-Trimethyl-Histone H4 (Lys20) antibody and the Magna ChIP™ A Kit (Cat. # 17-610).
Successful immunoprecipitation of trimethyl-histone H4 (Lys20)-associated DNA fragments was verified by qPCR using ChIP Primers B-Globin.
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.
Packaging Information
Material Size10 assays
Material Package10 assays per set. Recommended use: ~10 μL antibody per chromatin immunoprecipitation (dependent upon biological context).
Global Trade Item Number
Catalogue Number GTIN
17-671 04053252742231