17-614
ChIPAb+ Trimethyl-Histone H3 (Lys4) - ChIP Validated Antibody and Primer Set, rabbit monoclonal
from rabbit
Synonim(y):
Chip Rabbit Antibody and primer set, H3K4me3 ChIP, H3K4me3, Histone H3 (tri methyl K4), Histone H3K4me3, Histone H3K4me3 ChIP
Zaloguj sięWyświetlanie cen organizacyjnych i kontraktowych
About This Item
Kod UNSPSC:
12352203
eCl@ss:
32160702
NACRES:
NA.32
Polecane produkty
pochodzenie biologiczne
rabbit
Poziom jakości
klon
monoclonal
reaktywność gatunkowa
mouse, human
reaktywność gatunkowa (przewidywana na podstawie homologii)
mammals
producent / nazwa handlowa
ChIPAb+
Upstate®
metody
ChIP: suitable (ChIP-seq)
western blot: suitable
izotyp
IgG
numer dostępu NCBI
numer dostępu UniProt
Warunki transportu
dry ice
informacje o genach
human ... H3F3B(3021)
Opis ogólny
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.
Specyficzność
Trimethyl-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
Zastosowanie
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 A (Cat. # 17-610) Kit. Immunoprecipitation of trimethyl histone H3 (Lys4) associated DNA fragments was verified by qPCR using ChIP Primers p21 flanking the human p21 promoter that contains an Sp1 binding site (Please see figures).
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 (Cat. # 17-408) or EZ-ChIP (Cat. #17-371) protocol for experimental details.
ChIP-seq Analysis:
Chromatin immunoprecipitation was performed using the Magna ChIP HiSens kit (17-10460), 3 µL anti-trimethyl-Histone H3 (Lys4) antibody (cat# 17-614) or, 20 µL Protein A/G beads, and 1e6 crosslinked HeLa cell chromatin followed by DNA purification using magnetic beads. Libraries were prepared from Input and ChIP DNA samples using standard protocols with Illumina barcoded adapters, and analyzed on Illumina HiSeq instrument. An excess of eighteen million reads from FastQ files were mapped using Bowtie (http://bowtie-bio.sourceforge.net/manual.shtml) following TagDust (http://genome.gsc.riken.jp/osc/english/dataresource/) tag removal. Peaks were identified using MACS (http://luelab.dfci.harvard.edu/MACS/), with peaks and reads visualized as a custom track in UCSC Genome Browser (http://genome.ucsc.edu) from BigWig and BED files. The highest 25% of peaks identified in the 17-614 and 07-473 datasets showed 99% overlap with peaks identified in the ENCODE H3K4me3 BROAD Histone track for HeLa S3.
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 A (Cat. # 17-610) Kit. Immunoprecipitation of trimethyl histone H3 (Lys4) associated DNA fragments was verified by qPCR using ChIP Primers p21 flanking the human p21 promoter that contains an Sp1 binding site (Please see figures).
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 (Cat. # 17-408) or EZ-ChIP (Cat. #17-371) protocol for experimental details.
ChIP-seq Analysis:
Chromatin immunoprecipitation was performed using the Magna ChIP HiSens kit (17-10460), 3 µL anti-trimethyl-Histone H3 (Lys4) antibody (cat# 17-614) or, 20 µL Protein A/G beads, and 1e6 crosslinked HeLa cell chromatin followed by DNA purification using magnetic beads. Libraries were prepared from Input and ChIP DNA samples using standard protocols with Illumina barcoded adapters, and analyzed on Illumina HiSeq instrument. An excess of eighteen million reads from FastQ files were mapped using Bowtie (http://bowtie-bio.sourceforge.net/manual.shtml) following TagDust (http://genome.gsc.riken.jp/osc/english/dataresource/) tag removal. Peaks were identified using MACS (http://luelab.dfci.harvard.edu/MACS/), with peaks and reads visualized as a custom track in UCSC Genome Browser (http://genome.ucsc.edu) from BigWig and BED files. The highest 25% of peaks identified in the 17-614 and 07-473 datasets showed 99% overlap with peaks identified in the ENCODE H3K4me3 BROAD Histone track for HeLa S3.
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.
Opakowanie
25 assays per kit, ~3μL per chromatin immunoprecipitation
Komponenty
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
Jakość
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 protocol for experimental details.
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 protocol for experimental details.
Opis wartości docelowych
17 kDa
Postać fizyczna
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
Przechowywanie i stabilność
Stable for 1 year at -20°C from date of receipt
Komentarz do analizy
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.
Informacje prawne
UPSTATE is a registered trademark of Merck KGaA, Darmstadt, Germany
Oświadczenie o zrzeczeniu się odpowiedzialności
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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Kod klasy składowania
10 - Combustible liquids
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