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Key Documents

17-613

Sigma-Aldrich

ChIPAb+ p53 - ChIP Validated Antibody and Primer Set

from mouse

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About This Item

Kod UNSPSC:
12352203
eCl@ss:
32160702
NACRES:
NA.52

pochodzenie biologiczne

mouse

Poziom jakości

klon

monoclonal

reaktywność gatunkowa

human

producent / nazwa handlowa

ChIPAb+
Upstate®

metody

ChIP: suitable
western blot: suitable

izotyp

IgG

numer dostępu NCBI

numer dostępu UniProt

Warunki transportu

dry ice

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+ p53 set includes the p53 antibody raised against recombinant human p53, the negative control antibody (mouse normal IgG), and qPCR primers flanking an Sp1 binding site (to which p53 can be recruited1) within the human p21 (WAF1/CIP1/CDKN1A) promoter, amplifying a 105 base pair PCR product. The p53 and negative control antibodies are supplied in a scalable "per ChIP" reaction size and can be used to functionally validate the precipitation of p53 associated chromatin.

Specyficzność

Recognizes p53 in wild type and mutant forms of human origin.

Zastosowanie

Chromatin Immunoprecipitation Analysis:
Sonicated chromatin prepared from untreated or actinomycin D treated (30 ng/mL, 6 hrs.) U2OS cells (3 X 106 cell equivalents per IP) was subjected to chromatin immunoprecipitation using 1 μg of mouse Anti-p53 and the Magna ChIP® G (Cat. # 17-611) Kit. Immunoprecipitation of p53 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. As previously published, p53 association with this region of the p21 promoter increases upon actinomycin D treatment.

Chromatin Immunoprecipitation Analysis:
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 1 μg of mouse Anti-p53 and the Magna ChIP A (Cat. # 17-610) Kit.
Immunoprecipitation of p53 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. p53 immunoprecipitable activity associated with this promoter increases with UV treatment as observed in other studies.
Research Category
Epigenetics & Nuclear Function
Research Sub Category
Chromatin Biology
This ChIPAb+ p53 -ChIP Validated Antibody & Primer Set conveniently includes the antibody & the specific control PCR primers.

Opakowanie

25 assays per kit. ~1 μg per chromatin immunoprecipitation

Jakość

Routinely evaluated by chromatin immunoprecipitation and immunoblot on RIPA lysate of human Raji cells.

Opis wartości docelowych

53 kDa

Postać fizyczna

Anti-p53 (mouse monoclonal IgG). One vial containing 25 μg of of protein A purified mouse IgG2a in 25 μL of storage buffer (PBS, pH 7.4, containing 0.05% sodium azide and 1 mg/mL BSA). Store at -20°C.

Normal Mouse IgG. One vial containing 25 μg of normal mouse IgG in 25 μL volume. Store at -20°C.

ChIP Primers p21. One vial containing 75 μL of 5 μM of each primer specific for a region of the human p21 (WAF1/CIP1/CDKN1A) promoter. Store at -20°C.
FOR: GTG GCT CTG ATT GGC TTT CTG
REV: CTG AAA ACA GGC AGC CCA AG
Format: Purified
Protein A purified

Przechowywanie i stabilność

Stable for 1 year at -20°C from date of receipt

Komentarz do analizy

Control
Included negative control antibody mouse normal IgG and control primers specific for human p21 (WAF1/CIP1/CDKN1A) promoter.

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

10 - Combustible liquids


Certyfikaty analizy (CoA)

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

Odwiedź Bibliotekę dokumentów

Neha Parikh et al.
The Journal of pathology, 232(5), 522-533 (2014-01-01)
Mutations in the TP53 tumour suppressor gene occur in half of all human cancers, indicating its critical importance in inhibiting cancer development. Despite extensive studies, the mechanisms by which mutant p53 enhances tumour progression remain only partially understood. Here, using
Yap Ching Chew et al.
The Journal of biological chemistry, 287(20), 16168-16178 (2012-03-20)
Sulforaphane (SFN) is an important cancer preventive agent derived from cruciferous vegetables. We show that SFN treatment suppresses normal human keratinocyte proliferation via a mechanism that involves increased expression of p21(Cip1). SFN treatment produces a concentration-dependent increase in p21(Cip1) promoter
Li Gao et al.
The Journal of biological chemistry, 293(10), 3780-3792 (2018-01-24)
The most frequently used oral anti-coagulant warfarin has been implicated in inducing calcification of aortic valve interstitial cells (AVICs), whereas the mechanism is not fully understood. The low-level activation of p53 is found to be involved in osteogenic transdifferentiation and
Gengze Wu et al.
Circulation, 130(17), 1452-1465 (2014-08-27)
Long noncoding RNAs (lncRNAs) have recently been implicated in many biological processes and diseases. Atherosclerosis is a major risk factor for cardiovascular disease. However, the functional role of lncRNAs in atherosclerosis is largely unknown. We identified lincRNA-p21 as a key
L Sauer et al.
Oncogene, 29(18), 2628-2637 (2010-03-02)
Early growth response-1 (Egr-1) is overexpressed in human prostate tumors and contributes to cancer progression. On the other hand, mutation of p53 is associated with advanced prostate cancer, as well as with metastasis and hormone independence. This study shows that

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