chip-it ffpe chromatin preparation ii  (Active Motif)

Journal: International Journal of Molecular Sciences

Article Title: The Current State of Chromatin Immunoprecipitation (ChIP) from FFPE Tissues

doi: 10.3390/ijms23031103

Figure Lengend Snippet: Timeline of the FFPE-ChIP procedures and kits introduced over time. Procedures are in rectangles while kits are in ovals. Specific colors are used for each research group and company.

Article Snippet: PAT-ChIP and EPAT-ChIP, together with the procedure described for the commercial kit ChIP-IT FFPE Chromatin Preparation II from Active Motif, are the only procedures that involve the use of a “canonical” probe sonicator ( ) [ , ].

Techniques:

Journal: International Journal of Molecular Sciences

Article Title: The Current State of Chromatin Immunoprecipitation (ChIP) from FFPE Tissues

doi: 10.3390/ijms23031103

Figure Lengend Snippet: Overview of the FFPE-ChIP procedures and kits analyzed in this review.

Article Snippet: PAT-ChIP and EPAT-ChIP, together with the procedure described for the commercial kit ChIP-IT FFPE Chromatin Preparation II from Active Motif, are the only procedures that involve the use of a “canonical” probe sonicator ( ) [ , ].

Techniques:

Journal: International Journal of Molecular Sciences

Article Title: The Current State of Chromatin Immunoprecipitation (ChIP) from FFPE Tissues

doi: 10.3390/ijms23031103

Figure Lengend Snippet: Schematic overview of the main steps shared by the different procedures described in this review. FFPE tissue sections can be collected from the block as a whole or dissected (macro- or micro-) to enrich the starting material of specific tissue components. The sections are deparaffinized and rehydrated prior to be subjected to limited reversal of crosslinking by heat and chromatin extraction and shearing by focused ultrasonication or canonical probe sonication. Then, chromatin should be checked prior to proceed with immunoselection: an aliquot of chromatin is taken and decrosslinked, and the DNA is purified, fluorimetrically quantified, and separated by electrophoresis to evaluate the size of fragments. Chromatin is then immunoselected using antibodies directed against the protein or histone modification of interest. After capture of chromatin–antibody complexes and their washing to remove the nonspecific chromatin binding, selected chromatin is decrosslinked and purified using spin-column strategies. Finally, a preliminary evaluation of the specificity of the immunoselection by real-time qPCR should be performed prior to library preparation and NGS sequencing.

Article Snippet: PAT-ChIP and EPAT-ChIP, together with the procedure described for the commercial kit ChIP-IT FFPE Chromatin Preparation II from Active Motif, are the only procedures that involve the use of a “canonical” probe sonicator ( ) [ , ].

Techniques: Blocking Assay, Extraction, Sonication, Purification, Electrophoresis, Modification, Binding Assay, Sequencing

Journal: International Journal of Molecular Sciences

Article Title: The Current State of Chromatin Immunoprecipitation (ChIP) from FFPE Tissues

doi: 10.3390/ijms23031103

Figure Lengend Snippet: Summary of the procedural steps used by the different FFPE-ChIP techniques and kits for tissue processing before chromatin isolation.

Article Snippet: PAT-ChIP and EPAT-ChIP, together with the procedure described for the commercial kit ChIP-IT FFPE Chromatin Preparation II from Active Motif, are the only procedures that involve the use of a “canonical” probe sonicator ( ) [ , ].

Techniques: Isolation, Lysis, Sonication, Homogenization

Journal: International Journal of Molecular Sciences

Article Title: The Current State of Chromatin Immunoprecipitation (ChIP) from FFPE Tissues

doi: 10.3390/ijms23031103

Figure Lengend Snippet: Summary of the procedural steps used by the different FFPE-ChIP procedures for chromatin immunoselection and DNA purification.

Article Snippet: PAT-ChIP and EPAT-ChIP, together with the procedure described for the commercial kit ChIP-IT FFPE Chromatin Preparation II from Active Motif, are the only procedures that involve the use of a “canonical” probe sonicator ( ) [ , ].

Techniques: Purification, Recombinant, Magnetic Beads, DNA Purification

Journal: International Journal of Molecular Sciences

Article Title: The Current State of Chromatin Immunoprecipitation (ChIP) from FFPE Tissues

doi: 10.3390/ijms23031103

Figure Lengend Snippet: Timeline of the FFPE-ChIP procedures and kits introduced over time. Procedures are in rectangles while kits are in ovals. Specific colors are used for each research group and company.

Article Snippet: A first version, named ChIP-IT FFPE Chromatin Preparation (Active Motif, Carlsbad, CA, USA, cat #53030), was marketed by Active Motif from 2014 and joined in 2018 by a second version (ChIP-IT FFPE Chromatin Preparation II, Active Motif, cat #53031— and ).

Techniques:

Journal: International Journal of Molecular Sciences

Article Title: The Current State of Chromatin Immunoprecipitation (ChIP) from FFPE Tissues

doi: 10.3390/ijms23031103

Figure Lengend Snippet: Overview of the FFPE-ChIP procedures and kits analyzed in this review.

Article Snippet: A first version, named ChIP-IT FFPE Chromatin Preparation (Active Motif, Carlsbad, CA, USA, cat #53030), was marketed by Active Motif from 2014 and joined in 2018 by a second version (ChIP-IT FFPE Chromatin Preparation II, Active Motif, cat #53031— and ).

Techniques:

Journal: International Journal of Molecular Sciences

Article Title: The Current State of Chromatin Immunoprecipitation (ChIP) from FFPE Tissues

doi: 10.3390/ijms23031103

Figure Lengend Snippet: Schematic overview of the main steps shared by the different procedures described in this review. FFPE tissue sections can be collected from the block as a whole or dissected (macro- or micro-) to enrich the starting material of specific tissue components. The sections are deparaffinized and rehydrated prior to be subjected to limited reversal of crosslinking by heat and chromatin extraction and shearing by focused ultrasonication or canonical probe sonication. Then, chromatin should be checked prior to proceed with immunoselection: an aliquot of chromatin is taken and decrosslinked, and the DNA is purified, fluorimetrically quantified, and separated by electrophoresis to evaluate the size of fragments. Chromatin is then immunoselected using antibodies directed against the protein or histone modification of interest. After capture of chromatin–antibody complexes and their washing to remove the nonspecific chromatin binding, selected chromatin is decrosslinked and purified using spin-column strategies. Finally, a preliminary evaluation of the specificity of the immunoselection by real-time qPCR should be performed prior to library preparation and NGS sequencing.

Article Snippet: A first version, named ChIP-IT FFPE Chromatin Preparation (Active Motif, Carlsbad, CA, USA, cat #53030), was marketed by Active Motif from 2014 and joined in 2018 by a second version (ChIP-IT FFPE Chromatin Preparation II, Active Motif, cat #53031— and ).

Techniques: Blocking Assay, Extraction, Sonication, Purification, Electrophoresis, Modification, Binding Assay, Sequencing

Journal: International Journal of Molecular Sciences

Article Title: The Current State of Chromatin Immunoprecipitation (ChIP) from FFPE Tissues

doi: 10.3390/ijms23031103

Figure Lengend Snippet: Summary of the procedural steps used by the different FFPE-ChIP techniques and kits for tissue processing before chromatin isolation.

Article Snippet: A first version, named ChIP-IT FFPE Chromatin Preparation (Active Motif, Carlsbad, CA, USA, cat #53030), was marketed by Active Motif from 2014 and joined in 2018 by a second version (ChIP-IT FFPE Chromatin Preparation II, Active Motif, cat #53031— and ).

Techniques: Isolation, Lysis, Sonication, Homogenization

Journal: International Journal of Molecular Sciences

Article Title: The Current State of Chromatin Immunoprecipitation (ChIP) from FFPE Tissues

doi: 10.3390/ijms23031103

Figure Lengend Snippet: Summary of the procedural steps used by the different FFPE-ChIP procedures for chromatin immunoselection and DNA purification.

Article Snippet: A first version, named ChIP-IT FFPE Chromatin Preparation (Active Motif, Carlsbad, CA, USA, cat #53030), was marketed by Active Motif from 2014 and joined in 2018 by a second version (ChIP-IT FFPE Chromatin Preparation II, Active Motif, cat #53031— and ).

Techniques: Purification, Recombinant, Magnetic Beads, DNA Purification