High throughput automated chromatin immunoprecipitation as a platform for drug screening and antibody validation.

Lab on a Chip
Angela R WuStephen R Quake

Abstract

Chromatin immunoprecipitation (ChIP) is an assay for interrogating protein-DNA interactions that is increasingly being used for drug target discovery and screening applications. Currently the complexity of the protocol and the amount of hands-on time required for this assay limits its use to low throughput applications; furthermore, variability in antibody quality poses an additional obstacle in scaling up ChIP for large scale screening purposes. To address these challenges, we report HTChIP, an automated microfluidic-based platform for performing high-throughput ChIP screening measurements of 16 different targets simultaneously, with potential for further scale-up. From chromatin to analyzable PCR results only takes one day using HTChIP, as compared to several days up to one week for conventional protocols. HTChIP can also be used to test multiple antibodies and select the best performer for downstream ChIP applications, saving time and reagent costs of unsuccessful ChIP assays as a result of poor antibody quality. We performed a series of characterization assays to demonstrate that HTChIP can rapidly and accurately evaluate the epigenetic states of a cell, and that it is sensitive enough to detect the changes in the epigeneti...Continue Reading

References

Dec 17, 1997·Current Topics in Developmental Biology·J A Thomson, V S Marshall
Nov 6, 1998·Science·J A ThomsonJ M Jones
Apr 5, 2000·Nature Biotechnology·B E ReubinoffA Bongso
Aug 9, 2003·Science·Shiv I S Grewal, Danesh Moazed
Feb 21, 2004·The Journal of Biological Chemistry·Feng MiaoRama Natarajan
Sep 17, 2004·Genes & Development·Matthew S Hayden, Sankar Ghosh
Oct 27, 2004·Proceedings of the National Academy of Sciences of the United States of America·Jen-Chywan WangKeith R Yamamoto
Sep 21, 2005·Nature Methods·Gerald SchattenRoger Pedersen
Feb 2, 2006·Analytical Chemistry·Joshua S MarcusStephen R Quake
Apr 29, 2006·Analytical Chemistry·Joshua S MarcusStephen R Quake
Feb 27, 2007·Cell·Tony Kouzarides
Mar 7, 2007·Nature Reviews. Genetics·Manel Esteller
Apr 5, 2007·Nature Protocols·Joel D NelsonKarol Bomsztyk
Jun 1, 2007·Nature·Simmie L FosterRuslan Medzhitov
Jun 16, 2007·Frontiers in Bioscience : a Journal and Virtual Library·John Arne Dahl, Philippe Collas
Jul 21, 2007·Proceedings of the National Academy of Sciences of the United States of America·Joanne L AttemaIrving L Weissman
Oct 17, 2007·Annual Review of Medicine·Einav Nili Gal-YamPeter A Jones
Oct 19, 2007·PLoS Biology·Yan WangNigel S Atkinson
Oct 26, 2007·Nature·John Whitehead
Jan 19, 2008·Nucleic Acids Research·John Arne Dahl, Philippe Collas
Feb 7, 2008·BioTechniques·Luis G AcevedoPeggy J Farnham
Mar 14, 2008·The New England Journal of Medicine·Manel Esteller
Jun 10, 2008·Nature Protocols·John Arne Dahl, Philippe Collas
May 7, 2009·Lab on a Chip·Angela R WuStephen R Quake
Sep 9, 2009·Nature Reviews. Genetics·Peter J Park

❮ Previous
Next ❯

Citations

Nov 13, 2012·Analytical Chemistry·Michelle L KovarikNancy L Allbritton
Oct 5, 2013·Nature Nanotechnology·Carlos A Aguilar, Harold G Craighead
Aug 22, 2013·Biomicrofluidics·Toshiki MatsuokaShuichi Takayama
Feb 8, 2014·Lab on a Chip·Michael Junkin, Savaş Tay
Mar 6, 2013·Biomicrofluidics·Aaron M Streets, Yanyi Huang
May 15, 2013·Lab on a Chip·Mark D BorysiakJonathan D Posner
Jan 14, 2016·Biomedical Microdevices·Alejandro StarkTza-Huei Wang
Jul 28, 2015·Nature Methods·Zhenning CaoChang Lu
Sep 11, 2014·Methods : a Companion to Methods in Enzymology·Byung-Ryool HyunPaul D Soloway
Dec 19, 2013·Lab on a Chip·Jose L Garcia-Cordero, Sebastian J Maerkl
Sep 18, 2014·Cellular and Molecular Life Sciences : CMLS·Samuel BeckJonghwan Kim
Jun 13, 2013·Journal of Laboratory Automation·M Kursad ArazAmy E Herr
Nov 19, 2013·Journal of Biomolecular Screening·Oliver J DresslerAndrew J deMello
Jul 25, 2019·Lab on a Chip·Chengyu DengChang Lu
Aug 5, 2020·Cold Spring Harbor Protocols·James DeCaprio, Thomas O Kohl
Feb 22, 2017·Clinical and Translational Medicine·Jian Wang, Yuanlin Song
Mar 8, 2021·Genomics·Nasibeh ChenaraniJavad Zahiri
Jan 20, 2021·ACS Biomaterials Science & Engineering·Tobias MieslerLorenz Meinel
Dec 25, 2015·Cold Spring Harbor Protocols·Angela R Wu, Stephen R Quake

❮ Previous
Next ❯

Related Concepts

Trending Feeds

COVID-19

Coronaviruses encompass a large family of viruses that cause the common cold as well as more serious diseases, such as the ongoing outbreak of coronavirus disease 2019 (COVID-19; formally known as 2019-nCoV). Coronaviruses can spread from animals to humans; symptoms include fever, cough, shortness of breath, and breathing difficulties; in more severe cases, infection can lead to death. This feed covers recent research on COVID-19.

Blastomycosis

Blastomycosis fungal infections spread through inhaling Blastomyces dermatitidis spores. Discover the latest research on blastomycosis fungal infections here.

Nuclear Pore Complex in ALS/FTD

Alterations in nucleocytoplasmic transport, controlled by the nuclear pore complex, may be involved in the pathomechanism underlying multiple neurodegenerative diseases including Amyotrophic Lateral Sclerosis and Frontotemporal Dementia. Here is the latest research on the nuclear pore complex in ALS and FTD.

Applications of Molecular Barcoding

The concept of molecular barcoding is that each original DNA or RNA molecule is attached to a unique sequence barcode. Sequence reads having different barcodes represent different original molecules, while sequence reads having the same barcode are results of PCR duplication from one original molecule. Discover the latest research on molecular barcoding here.

Chronic Fatigue Syndrome

Chronic fatigue syndrome is a disease characterized by unexplained disabling fatigue; the pathology of which is incompletely understood. Discover the latest research on chronic fatigue syndrome here.

Evolution of Pluripotency

Pluripotency refers to the ability of a cell to develop into three primary germ cell layers of the embryo. This feed focuses on the mechanisms that underlie the evolution of pluripotency. Here is the latest research.

Position Effect Variegation

Position Effect Variagation occurs when a gene is inactivated due to its positioning near heterochromatic regions within a chromosome. Discover the latest research on Position Effect Variagation here.

STING Receptor Agonists

Stimulator of IFN genes (STING) are a group of transmembrane proteins that are involved in the induction of type I interferon that is important in the innate immune response. The stimulation of STING has been an active area of research in the treatment of cancer and infectious diseases. Here is the latest research on STING receptor agonists.

Microbicide

Microbicides are products that can be applied to vaginal or rectal mucosal surfaces with the goal of preventing, or at least significantly reducing, the transmission of sexually transmitted infections. Here is the latest research on microbicides.