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17-651 ChIPAb+ Monomethyl-Histone H4 (Lys20) - ChIP Validated Antibody and Primer Set

This ChIPAb+ Monomethyl-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
17-651
25 assays  25 assays per set. Recommended use: ~4 μg antibody per chromatin immunoprecipitation (dependent upon biological context).
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Key Spec Table

Species ReactivityKey Applications
H, Pl, VrtWB, ChIP
Description
Catalogue Number17-651
Brand Family Upstate
Trade Name
  • ChIPAb+
  • Upstate
DescriptionChIPAb+ Monomethyl-Histone H4 (Lys20) - 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 H4 (Lys20) set includes the Anti-monomethyl-Histone H4 (Lys20) antibody, a negative control antibody (purified Rabbit IgG), and qPCR primers, which amplify a 212 base pair PCR product within the human GAPDH coding region. The monomethyl 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 monomethyl-histone H4 (Lys20) associated chromatin.
Alternate Names
  • H4K20me1
  • Histone H4 (mono methyl K20)
  • H4 histone family, member A
  • histone 1
  • H4a
  • histone cluster 1
  • H4a
Product Information
FormatAffinity Purified
Control
  • Includes negative control rabbit IgG antibody and control primers specific for human GAPDH coding region.
PresentationAnti-monomethyl-Histone H4 (Lys20) (rabbit polyclonal IgG). One vial containing 100 μg of affinity purified antibody in 100 μL 0.1M Tris-Glycine (pH 7.4), 15 mM NaCl, and 0.05% NaN3. Store at -20°C.

Normal Rabbit IgG. One vial containing 125 μg of purified rabbit IgG in 125 μL storage buffer. Store at -20°C.

ChIP Primers GAPDH Coding Region. One vial containing 75 μL of 5 μM of each primer specific for human GAPDH coding region.Store at -20°C.
FOR: GGC TCC CAC CTT TCT CAT CC
REV: GGC CAT CCA CAG TCT TCT GG
Quality LevelMQ100
Applications
ApplicationThis ChIPAb+ Monomethyl-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 4 μg of either a negative control antibody or Anti-Monomethyl-Histone H4
(Lys20) antibody and the Magna ChIP A Kit (Cat. #17-610). Successful immunoprecipitation of monomethyl-histone H4 (Lys20)-associated DNA fragments was verified by qPCR using GAPDH coding region 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 antibody shown as (-) and monomethyl-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:
0.1 μg/mL of proteins were visualized using a goat anti-rabbit secondary antibody conjugated to HRP and a chemiluminescence detection system (Please see figures)

Peptide Blocking Assay:
40 μg of histone H4 peptide containing monomethyl Lys20 abolished detection of histone H4 by anti-monomethyl-Histone H4 (Lys20) in immunoblot analysis of acid extracts of HeLa cells (Please see figures).
Biological Information
ImmunogenThe immunogen was a synthetic peptide corresponding to amino acids 15-24 of histone H4, monomethylated on Lys20.
Epitopea.a. 15-24
HostRabbit
SpecificityRecognizes histone H4, Mr ~11 kDa.
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 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. 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 4 μg of either a negative control antibody or Anti-monomethyl-Histone H4 (Lys20) antibody and the Magna ChIP A Kit (Cat. # 17-610). Successful immuno-precipitation of monomethyl-histone H4 (Lys20)-associated DNA fragments was verified by qPCR using ChIP Primers GAPDH Coding region (Please see figures).
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 Size25 assays
Material Package25 assays per set. Recommended use: ~4 μg antibody per chromatin immunoprecipitation (dependent upon biological context).
Global Trade Item Number
Catalogue Number GTIN
17-651 04053252477522