Przejdź do zawartości
Merck
Wszystkie zdjęcia(4)

Key Documents

17-10245

Sigma-Aldrich

ChIPAb+ Histone H3.3 - ChIP Validated Antibody and Primer Set

from rabbit, purified by affinity chromatography

Synonim(y):

Histone H3.3

Zaloguj sięWyświetlanie cen organizacyjnych i kontraktowych


About This Item

Kod UNSPSC:
12352203
eCl@ss:
32160702
NACRES:
NA.32

pochodzenie biologiczne

rabbit

Poziom jakości

klon

polyclonal

oczyszczone przez

affinity chromatography

reaktywność gatunkowa

mouse, human

producent / nazwa handlowa

ChIPAb+
Upstate®

metody

ChIP: suitable
dot blot: suitable
western blot: suitable

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+ 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.

Specyficzność

Broad species cross-reactivity expected based on 100% sequence homology.
This antibody recognizes human Histone H3.

Immunogen

Epitope: a.a. 85-105
KLH-conjugated linear peptide corresponding to human Histone H3.3.

Zastosowanie

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.
Research Category
Epigenetics & Nuclear Function
Research Sub Category
Histones
This ChIPAb+ Histone H3.3 -ChIP Validated Antibody & Primer Set conveniently includes the antibody & the specific control PCR primers.

Opakowanie

25 assays per set. Recommended use: ~2 μg of antibody per chromatin immunoprecipitation (dependent upon biological context).

Jakość

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.

Opis wartości docelowych

~17 kDa observed

Postać fizyczna

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

Przechowywanie i stabilność

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.

Komentarz do analizy

Control
Includes normal rabbit IgG and primers specific for human GAPDH coding region.

Inne uwagi

Concentration: Please refer to the Certificate of Analysis for the lot-specific concentration.

Informacje prawne

MAGNA CHIP is a registered trademark of Merck KGaA, Darmstadt, Germany
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.
This page may contain text that has been machine translated.

Kod klasy składowania

12 - Non Combustible Liquids

Temperatura zapłonu (°F)

Not applicable

Temperatura zapłonu (°C)

Not applicable


Certyfikaty analizy (CoA)

Poszukaj Certyfikaty analizy (CoA), wpisując numer partii/serii produktów. Numery serii i partii można znaleźć na etykiecie produktu po słowach „seria” lub „partia”.

Masz już ten produkt?

Dokumenty związane z niedawno zakupionymi produktami zostały zamieszczone w Bibliotece dokumentów.

Odwiedź Bibliotekę dokumentów

Analysis of neonatal brain lacking ATRX or MeCP2 reveals changes in nucleosome density, CTCF binding and chromatin looping.
Kernohan, KD; Vernimmen, D; Gloor, GB; Berube, NG
Nucleic Acids Research null
William Chang et al.
Epigenetics & chromatin, 16(1), 4-4 (2023-01-27)
Cellular differentiation is marked by temporally and spatially coordinated gene expression regulated at multiple levels. DNA methylation represents a universal mechanism to control chromatin organization and its accessibility. Cytosine methylation of CpG dinucleotides regulates binding of methylation-sensitive DNA-binding transcription factors
Joseph Aaron Goldman et al.
Developmental cell, 40(4), 392-404 (2017-03-02)
Chromatin regulation is a principal mechanism governing animal development, yet it is unclear to what extent structural changes in chromatin underlie tissue regeneration. Non-mammalian vertebrates such as zebrafish activate cardiomyocyte (CM) division after tissue damage to regenerate lost heart muscle.
Liquan Zhou et al.
Development (Cambridge, England), 144(3), 519-528 (2016-12-21)
During development from oocyte to embryo, genetic programs in mouse germ cells are reshaped by chromatin remodeling to orchestrate the onset of development. Epigenetic modifications of specific amino acid residues of core histones and their isoforms can dramatically alter activation
Renee J Tamming et al.
Cell reports, 31(13), 107838-107838 (2020-07-02)
ATRX gene mutations have been identified in syndromic and non-syndromic intellectual disabilities in humans. ATRX is known to maintain genomic stability in neuroprogenitor cells, but its function in differentiated neurons and memory processes remains largely unresolved. Here, we show that

Nasz zespół naukowców ma doświadczenie we wszystkich obszarach badań, w tym w naukach przyrodniczych, materiałoznawstwie, syntezie chemicznej, chromatografii, analityce i wielu innych dziedzinach.

Skontaktuj się z zespołem ds. pomocy technicznej