17-614
ChIPAb+ Trimethyl-Histone H3 (Lys4) - ChIP Validated Antibody and Primer Set, rabbit monoclonal
from rabbit
Synonym(s):
Chip Rabbit Antibody and primer set, H3K4me3 ChIP, H3K4me3, Histone H3 (tri methyl K4), Histone H3K4me3, Histone H3K4me3 ChIP
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| Size/SKU | Availability | Price |
|---|---|---|
25 assays | Check Cart for Availability | $634.00 |
About This Item
UNSPSC Code:
12352203
NACRES:
NA.32
eCl@ss:
32160702
Clone:
monoclonal
Species reactivity:
mouse, human
Application:
ChIP, WB
Citations:
183
biological source
rabbit
Quality Segment
clone
monoclonal
species reactivity
mouse, human
species reactivity (predicted by homology)
mammals
manufacturer/tradename
ChIPAb+, Upstate®
technique(s)
ChIP: suitable (ChIP-seq), western blot: suitable
isotype
IgG
NCBI accession no.
UniProt accession no.
shipped in
dry ice
Gene Information
human ... H3F3B(3021)
General description
17 kDa
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+ Trimethyl-Histone H3 (Lys4) set includes the anti-trimethyl-histone H3 (Lys4) antibody, a negative control antibody (normal rabbit serum), and qPCR primers which amplify a 166 base pair region within the promoter of the human GAPDH gene. The trimethyl-histone H3 (Lys4) 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 H3 (Lys4)-associated chromatin.
The ChIPAb+ Trimethyl-Histone H3 (Lys4) set includes the anti-trimethyl-histone H3 (Lys4) antibody, a negative control antibody (normal rabbit serum), and qPCR primers which amplify a 166 base pair region within the promoter of the human GAPDH gene. The trimethyl-histone H3 (Lys4) 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 H3 (Lys4)-associated chromatin.
The methylation of histones can occur on two different residues: arginine or lysine. Histone methylation can be associated with transcriptional activation or repression, depending on the methylated residue. Lysine 4 of histone H3 can be mono-, di- or trimethylated by different histone methyltransferases (HMTs) such as SET1 or ASH1. Methylation of Lys4 is often associated with transcriptional activation. The demethylase LSD1 is able to demethylate histone H3 Lys4.
Immunogen
Epitope: Trimethyl Lys4
The trimethyl-histone H3 (Lys4) antibody is made against a BSA-conjugated, synthetic peptide containing the sequence …RT[me3K]QT… in which me3K corresponds to trimethyl-lysine 4 of human histone H3
Application
Chromatin Immunoprecipitation:
Sonicated chromatin prepared from untreated or UV treated (6 hrs, 50 joules/m2.) U2OS cells (3 X 106 cell equivalents per IP) was subjected to chromatin immunoprecipitation using 3 μL of rabbit Anti-Trimethyl Histone H3 (Lys4) and the Magna ChIP
Fold Increase is a ratio of normalized mean IP quantities extracted from standard curves derived from inputs of each chromatin sample. Trimethyl-histone (Lys4) immunoprecipitable activity associated with this promoter increases with UV treatment as observed in other studies.
Please refer to the EZ-Magna A ChIP
ChIP-seq Analysis:
Chromatin immunoprecipitation was performed using the Magna ChIP
Western blot analysis and peptide inhibition:
Representative blot. HeLa acid extract was resolved by electrophoresis, transferred to nitrocellulose and probed with anti-trimethyl-histone H3 (Lys4) (1:2,000, lane 1) or preincubated with 0.4 μM Histone H3 peptide with following modifications Lane 2: monomethyl-Lysine 4, Lane 3: dimethyl Lysine 4, Lane 4: trimethyl-Lysine 4.
Proteins were visualized using a goat anti-rabbit secondary antibody conjugated to HRP and a chemiluminescence detection
system (Please see figures).
Sonicated chromatin prepared from untreated or UV treated (6 hrs, 50 joules/m2.) U2OS cells (3 X 106 cell equivalents per IP) was subjected to chromatin immunoprecipitation using 3 μL of rabbit Anti-Trimethyl Histone H3 (Lys4) and the Magna ChIP
Fold Increase is a ratio of normalized mean IP quantities extracted from standard curves derived from inputs of each chromatin sample. Trimethyl-histone (Lys4) immunoprecipitable activity associated with this promoter increases with UV treatment as observed in other studies.
Please refer to the EZ-Magna A ChIP
ChIP-seq Analysis:
Chromatin immunoprecipitation was performed using the Magna ChIP
Western blot analysis and peptide inhibition:
Representative blot. HeLa acid extract was resolved by electrophoresis, transferred to nitrocellulose and probed with anti-trimethyl-histone H3 (Lys4) (1:2,000, lane 1) or preincubated with 0.4 μM Histone H3 peptide with following modifications Lane 2: monomethyl-Lysine 4, Lane 3: dimethyl Lysine 4, Lane 4: trimethyl-Lysine 4.
Proteins were visualized using a goat anti-rabbit secondary antibody conjugated to HRP and a chemiluminescence detection
system (Please see figures).
Research Category
Epigenetics & Nuclear Function
Epigenetics & Nuclear Function
Research Sub Category
Chromatin Biology
Chromatin Biology
Trimethyl-Histone H3 (Lys4) ChIP validated antibody & primer set including the ChIP-grade antibody & the specific control PCR primers used for chromatin immunoprecipitation of H3K4Me3.
Biochem/physiol Actions
Trimethyl-Histone H3 (Lys4)
Packaging
25 assays per kit, ~3μL per chromatin immunoprecipitation
Physical form
Anti-Trimethyl-Histone H3 (Lys4) recombin-ant rabbit monoclonal IgG. One vial containing 75 μL of protein A purified rabbit IgG in storage buffer (0.1M Tris-Glycine pH 7.4, 0.15M NaCl, 0.05% NaN3, with the addition of 40% glycerol.
Normal Rabbit IgG. One vial containing 75 μL of normal rabbit IgG.
Control Primers. One vial containing 75 μL of 5 μM of each primer specific for human GAPDH.
FOR: TAC TAG CGG TTT TAC GGG CG
REV: TCG AAC AGG AGG AGC AGA GAG CGA
Normal Rabbit IgG. One vial containing 75 μL of normal rabbit IgG.
Control Primers. One vial containing 75 μL of 5 μM of each primer specific for human GAPDH.
FOR: TAC TAG CGG TTT TAC GGG CG
REV: TCG AAC AGG AGG AGC AGA GAG CGA
Format: Purified
Preparation Note
Stable for 1 year at -20°C from date of receipt
Analysis Note
Chromatin Immunoprecipitation:
Sonicated chromatin prepared from untreated HeLa cells (1 X 106 cell equivalents) was subjected to chromatin immunoprecipitation using 3 μL of either a normal rabbit IgG or 3 μL Anti-Trimethyl-Histone H3 (Lys4) Monoclonal IgG and the Magna ChIP A (Part # 17-610) Kit. Successful immunoprecipitation of
trimethyl-histone H3 (Lys4) associated DNA fragments was verified by qPCR using control ChIP Primers flanking the human GAPDH promoter (Please see figures). Please refer to the EZ-Magna A ChIP
Sonicated chromatin prepared from untreated HeLa cells (1 X 106 cell equivalents) was subjected to chromatin immunoprecipitation using 3 μL of either a normal rabbit IgG or 3 μL Anti-Trimethyl-Histone H3 (Lys4) Monoclonal IgG and the Magna ChIP A (Part # 17-610) Kit. Successful immunoprecipitation of
trimethyl-histone H3 (Lys4) associated DNA fragments was verified by qPCR using control ChIP Primers flanking the human GAPDH promoter (Please see figures). Please refer to the EZ-Magna A ChIP
Control
Included negative control antibody purified rabbit IgG and control primers specific for human GAPDH promoter.
Included negative control antibody purified rabbit IgG and control primers specific for human GAPDH promoter.
Other Notes
Anti-Trimethyl-Histone H3 (Lys4) (purified rabbit IgG), 1 vial
Negative ChIP Control Rabbit IgG, 1 vial
ChIP Primers GAPDH, 1 vial
Negative ChIP Control Rabbit IgG, 1 vial
ChIP Primers GAPDH, 1 vial
Legal Information
UPSTATE is a registered trademark of Merck KGaA, Darmstadt, Germany
Disclaimer
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.
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This Item | |||
|---|---|---|---|
| description from rabbit | description from rabbit | description from rabbit, purified by using Protein A | description clone CMA304, from mouse, purified by using protein G |
| species reactivity mouse, human | species reactivity human, mouse, vertebrates | species reactivity human, mouse | species reactivity human, vertebrates |
| biological source rabbit | biological source rabbit | biological source rabbit | biological source mouse |
| clone monoclonal | clone polyclonal | clone polyclonal | clone CMA304, monoclonal |
| Gene Information human ... H3F3B(3021) | Gene Information human ... H3F3B(3021) | Gene Information human ... H3F3B(3021) | Gene Information - |
| UniProt accession no. | UniProt accession no. | UniProt accession no. | UniProt accession no. |
| technique(s) ChIP: suitable (ChIP-seq), western blot: suitable | technique(s) ChIP: suitable, dot blot: suitable, western blot: suitable | technique(s) ChIP: suitable, western blot: suitable | technique(s) ChIP: suitable (ChIP-seq), immunocytochemistry: suitable, western blot: suitable |
Storage Class
10 - Combustible liquids
wgk
WGK 1
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