17-10245
ChIPAb+ Histone H3.3 - ChIP Validated Antibody and Primer Set
from rabbit, purified by affinity chromatography
Sinônimo(s):
Histone H3.3
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About This Item
Código UNSPSC:
12352203
eCl@ss:
32160702
NACRES:
NA.32
Produtos recomendados
fonte biológica
rabbit
Nível de qualidade
clone
polyclonal
purificado por
affinity chromatography
reatividade de espécies
mouse, human
fabricante/nome comercial
ChIPAb+
Upstate®
técnica(s)
ChIP: suitable
dot blot: suitable
western blot: suitable
nº de adesão NCBI
nº de adesão UniProt
Condições de expedição
dry ice
Informações sobre genes
human ... H3F3B(3021)
Descrição geral
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+ Histone H3.3 set includes the Histone H3.3 antibody, a Normal rabbit IgG, and control primers which amplify a 87 bp region of ChIP Primers, GAPDH Coding Region D2 Human. The Histone H3.3 and negative controls are supplied in a scalable "per ChIP" reaction size and can be used to functionally validate the precipitation of Histone H3.3 -associated chromatin.
The ChIPAb+ Histone H3.3 set includes the Histone H3.3 antibody, a Normal rabbit IgG, and control primers which amplify a 87 bp region of ChIP Primers, GAPDH Coding Region D2 Human. The Histone H3.3 and negative controls are supplied in a scalable "per ChIP" reaction size and can be used to functionally validate the precipitation of Histone H3.3 -associated chromatin.
Especificidade
Broad species cross-reactivity expected based on 100% sequence homology.
This antibody recognizes human Histone H3.
Imunogênio
Epitope: a.a. 85-105
KLH-conjugated linear peptide corresponding to human Histone H3.3.
Aplicação
Chromatin Immunoprecipitation:
Representative lot data.
Sonicated chromatin prepared from HeLa cells (1 X 10E6 cell equivalents per IP) were subjected to chromatin immunoprecipitation using 2 µg of either Normal rabbit IgG or 2 µg Anti-Histone H3.3 and the Magna ChIP A Kit (Cat. # 17-610). Successful immunoprecipitation of Histone H3.3 associated DNA fragments was verified by qPCR using ChIP Primers, GAPDH Coding Region D2 as a positive locus, and beta-actin promoter primers 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 A (Cat. # 17-408) or EZ-ChIP (Cat. # 17-371) protocol for experimental details.
Western Blot Analysis:
Representative lot data.
HeLa acid extract was probed with Anti-Histone H3.3 (1 µg/mL). Proteins were visualized using a Donkey Anti-Rabbit IgG secondary antibody conjugated to HRP and a chemiluminescence detection system.
Arrow indicates Histone H3.3 (~17 kDa). (Figure 3).
Dot Blot Analysis;
Representative lot data.
Recombinant H3.2 and H3.3 spotted on PVDF (1, 10, 100, 1,000 ng histone per slot). Detection with 0.5 µg/mL a-H3.3 antibody in TBS-T, 5% milk. (Figure 4).
Image courtesy of Simon Elsässer, Laboratory of David Allis, Rockefeller University, New York.
Representative lot data.
Sonicated chromatin prepared from HeLa cells (1 X 10E6 cell equivalents per IP) were subjected to chromatin immunoprecipitation using 2 µg of either Normal rabbit IgG or 2 µg Anti-Histone H3.3 and the Magna ChIP A Kit (Cat. # 17-610). Successful immunoprecipitation of Histone H3.3 associated DNA fragments was verified by qPCR using ChIP Primers, GAPDH Coding Region D2 as a positive locus, and beta-actin promoter primers 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 A (Cat. # 17-408) or EZ-ChIP (Cat. # 17-371) protocol for experimental details.
Western Blot Analysis:
Representative lot data.
HeLa acid extract was probed with Anti-Histone H3.3 (1 µg/mL). Proteins were visualized using a Donkey Anti-Rabbit IgG secondary antibody conjugated to HRP and a chemiluminescence detection system.
Arrow indicates Histone H3.3 (~17 kDa). (Figure 3).
Dot Blot Analysis;
Representative lot data.
Recombinant H3.2 and H3.3 spotted on PVDF (1, 10, 100, 1,000 ng histone per slot). Detection with 0.5 µg/mL a-H3.3 antibody in TBS-T, 5% milk. (Figure 4).
Image courtesy of Simon Elsässer, Laboratory of David Allis, Rockefeller University, New York.
Research Category
Epigenetics & Nuclear Function
Epigenetics & Nuclear Function
Research Sub Category
Histones
Histones
This ChIPAb+ Histone H3.3 -ChIP Validated Antibody & Primer Set conveniently includes the antibody & the specific control PCR primers.
Embalagem
25 assays per set. Recommended use: ~2 μg of antibody per chromatin immunoprecipitation (dependent upon biological context).
Qualidade
Chromatin Immunoprecipitation:
Representative lot data.
Sonicated chromatin prepared from HeLa cells (1 X 10E6 cell equivalents per IP) were subjected to chromatin immunoprecipitation using 2 µg of either Normal Rabbit IgG or 2 µg of Anti-Histone H3.3 and the Magna ChIP® A Kit (Cat. # 17-610). Successful immunoprecipitation of Histone H3.3 associated DNA fragments was verified by qPCR using ChIP Primers, GAPDH Coding Region D2 (Figure 1).
Please refer to the EZ-Magna ChIP A (Cat. # 17-408) or EZ-ChIP (Cat. # 17-371) protocol for experimental details.
Representative lot data.
Sonicated chromatin prepared from HeLa cells (1 X 10E6 cell equivalents per IP) were subjected to chromatin immunoprecipitation using 2 µg of either Normal Rabbit IgG or 2 µg of Anti-Histone H3.3 and the Magna ChIP® A Kit (Cat. # 17-610). Successful immunoprecipitation of Histone H3.3 associated DNA fragments was verified by qPCR using ChIP Primers, GAPDH Coding Region D2 (Figure 1).
Please refer to the EZ-Magna ChIP A (Cat. # 17-408) or EZ-ChIP (Cat. # 17-371) protocol for experimental details.
Descrição-alvo
~17 kDa observed
forma física
Affinity purified
Anti-Histone H3.3 (rabbit polyclonal). One vial containing 50 µg of purified rabbit polyclonal in buffer containing 0.1M Tris-Glycine (pH 7.4), 150 mM NaCl with 0.05% sodium azide before the addition of glycerol to 30%. Store at -20° C.
Concentration: 0.7 mg/mL
Normal Rabbit IgG. One vial containing 125 µg of Rabbit IgG in 125 µL of storage buffer containing 0.05% sodium azide. Store at -20°C.
Control Primers, GAPDH Coding Region D2 Human. One vial containing 75 μL of 5 μM of each primer specific for human GAPDH coding region. Store at -20°C.
FOR: GCC ATG TAG ACC CCT TGA AGA G
REV: ACT GGT TGA GCA CAG GGT ACT TTA T
Concentration: 0.7 mg/mL
Normal Rabbit IgG. One vial containing 125 µg of Rabbit IgG in 125 µL of storage buffer containing 0.05% sodium azide. Store at -20°C.
Control Primers, GAPDH Coding Region D2 Human. One vial containing 75 μL of 5 μM of each primer specific for human GAPDH coding region. Store at -20°C.
FOR: GCC ATG TAG ACC CCT TGA AGA G
REV: ACT GGT TGA GCA CAG GGT ACT TTA T
Armazenamento e estabilidade
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.
Note: Variability in freezer temperatures below -20°C may cause glycerol containing solutions to become frozen during storage.
Nota de análise
Control
Includes normal rabbit IgG and primers specific for human GAPDH coding region.
Includes normal rabbit IgG and primers specific for human GAPDH coding region.
Outras notas
Concentration: Please refer to the Certificate of Analysis for the lot-specific concentration.
Informações legais
MAGNA CHIP is a registered trademark of Merck KGaA, Darmstadt, Germany
UPSTATE is a registered trademark of Merck KGaA, Darmstadt, Germany
Exoneração de responsabilidade
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.
Código de classe de armazenamento
12 - Non Combustible Liquids
Ponto de fulgor (°F)
Not applicable
Ponto de fulgor (°C)
Not applicable
Certificados de análise (COA)
Busque Certificados de análise (COA) digitando o Número do Lote do produto. Os números de lote e remessa podem ser encontrados no rótulo de um produto após a palavra “Lot” ou “Batch”.
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