半岛bd体育手机客户端 说明
一般描述
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 (Lys79) set includes the Trimethyl-Histone H3 (Lys79) antibody, a negative control normal rabbit IgG, and qPCR primers which amplify a 166 bp region of human GAPDH. The Trimethyl-Histone H3 (Lys79) and negative controls are supplied in a scalable "per ChIP" reaction size and can be used to functionally validate the precipitation of Trimethyl-Histone H3 (Lys79)-associated chromatin.
Histones are highly conserved proteins that serve as the structural scaffold for the organization of nuclear DNA into chromatin. The four core histones, H2A, H2B, H3, and H4, assemble into an octamer (2 molecules of each). Subsequently, 146 base pairs of DNA are wrapped around the octamer, forming a nucleosome, the basic subunit of chromatin. Histone modifications regulate DNA transcription, repair, recombination, and replication. The most commonly studied modifications are acetylation, phosphorylation, methylation, and ubiquitination. Lysine methylation occurs in three distinct states, having either one (me1), two (me2) or three (me3) methyl groups attached to the amine group of the lysine side chain. Methylation of Lysine 79 in Histone 3 acts as a marker of inactive chromatin regions. Methylation by Dot1 methylase on lysine-79 (Lys79) of Histone H3 is criticial for transcriptional silencing, and it is thought that silencing proteins such as Sir3 function by blocking Dot1 methylation.
特异性
This antibody recognizes Histone H3 Trimethylated on Lys79.
Broad species reactivity is expected.
免疫原
Linear Peptide corresponding to human Histone H3 methylated at Lys79.
Epitope: Trimethyl Lys79
应用
This ChIPAb+ Trimethyl-Histone H3 (Lys79) -ChIP Validated Antibody & Primer Set conveniently includes the antibody & the specific control PCR primers.
Chromatin Immunoprecipitation:
Representative lot data.
Sonicated chromatin prepared from HeLa cells (1 X 10E6 cell equivalents per IP) were subjected to chromatin immunoprecipitation using either 1 ?g of normal rabbit IgG, or 1 ?g of Anti-trimethyl-Histone H3 (Lys79) and the Magna ChIP?A Kit (Cat. # 17-610).
Successful immunoprecipitation of trimethyl-Histone H3 (Lys79) associated DNA fragments was verified by qPCR using Control Primers as a positive locus, and human β-globin as a negative locus (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
Western Blot Analysis:
Representative lot data.
HeLa acid extract (lane 1) and recombinant Histone H3 (Cat # 14-494) (lane2) lysates were resolved by electrophoresis, transferred to PVDF membrane and probed with Anti-trimethyl Histone H3 (Lys79) (0.1 μg/mL). Proteins were visualized using a Donkey Anti-rabbit IgG conjugated to HRP and a chemiluminescence detection system.
Arrow indicates Histone H3 (~17 kDa) (Figure 3).
Multiplex Immunoassay: Representative lot data.
Specificity of interaction of anti-trimethyl histone H3 Lys79 antibody was assayed using the Luminex
Research Category
Epigenetics & Nuclear Function
Research Sub Category
Histones
Chromatin Biology
包装
25 assays per set. Recommended use: ~1 μg of antibody per chromatin immunoprecipitation (dependent upon biological context).
质量
Chromatin Immunoprecipitation:
Sonicated chromatin prepared from HeLa cells (1 X 10E6 cell equivalents per IP) were subjected to chromatin immunoprecipitation using either 1 ?g of normal rabbit IgG, or 1 ?g of Anti-trimethyl-Histone H3 (Lys79) and the Magna ChIP?A Kit (Cat. # 17-610). Successful immunoprecipitation of trimethyl-Histone H3 (Lys79) associated DNA fragments was verified by qPCR using Control Primers, (Figure 1).
Please refer to the EZ-Magna ChIP
目标描述
~ 17 kDa
外形
Anti-Trimethyl-Histone H3 (Lys79) (rabbit polyclonal), Part No. CS204342. One vial containing 25 ?g of purified rabbit polyclonal 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, Part No. PP64B. One vial containing 125 ?g Rabbit IgG in 125 ?L storage buffer containing 0.05% sodium azide.
Store at -20°C.
Control Primers, Part No. 22-004. One vial containing 75 μL of 5 μM of 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
Affinity purified
储存及稳定性
Stable 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.
分析说明
Control
Includes negative control normal rabbit IgG and primers specific for human GAPDH.
其他说明
Concentration: Please refer to the Certificate of Analysis for the lot-specific concentration.
法律信息
MAGNA CHIP is a registered trademark of Merck KGaA, Darmstadt, Germany
UPSTATE is a registered trademark of Merck KGaA, Darmstadt, Germany
免责声明
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.
基本信息
| eCl@ss | 32160702 |
半岛bd体育手机客户端 性质
| 生物来源 | rabbit |
| 质量水平 | 100 |
| 克隆 | polyclonal |
| 纯化方式 | affinity chromatography |
| species reactivity | human, vertebrates |
| manufacturer/tradename | ChIPAb+ Upstate? |
| technique(s) | ChIP: suitable multiplexing: suitable western blot: suitable |
| NCBI登记号 | NP_003484.1 |
| UniProt登记号 | Q16695 |
| 运输 | dry ice |
安全信息
| 储存分类代码 | 10 - Combustible liquids |
Sigma-Aldrich
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