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Related Concept Videos

Immunoprecipitation01:20

Immunoprecipitation

7.9K
Immunoprecipitation, or IP, is a widely used technique that employs protein-antibody interactions to isolate proteins or protein complexes in their native state for studying protein-protein interactions, quaternary structures, or supramolecular complexes. Various modifications of the technique, including chromatin IP, cross-linking IP, and fluorescence IP, are commonly used.
Chromatin Immunoprecipitation
Chromatin immunoprecipitation, also known as ChIP, is used to study protein-DNA or...
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Chromatin Immunoprecipitation- ChIP02:36

Chromatin Immunoprecipitation- ChIP

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Chromatin immunoprecipitation, or ChIP, is an antibody-based technique used to identify sites on DNA that bind to transcription factors of interest or histone proteins. It also helps determine the type of histone modifications such as acetylation, phosphorylation, or methylation.
Types of ChIP
ChIP can be divided into two types - X-ChIP and N-ChIP. X-ChIP involves in vivo cross-linking of histones and regulatory proteins to DNA, fragmenting the DNA by sonication, and isolating the protein-DNA...
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Precipitation and Co-precipitation01:17

Precipitation and Co-precipitation

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Precipitation and coprecipitation methods can be used to separate a mixture of ions in a solution. In qualitative inorganic analysis, ions that form sparingly soluble precipitates with the same reagent are separated based on the differences in solubility products. For example, consider the separation of Cu(II) and Fe(II) ions by precipitation as insoluble sulfides. First, copper(II) sulfide is precipitated by the addition of acidic H2S, where the dissociation of H2S is suppressed. Adding H2S...
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Immunogold Electron Microscopy01:20

Immunogold Electron Microscopy

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Immunoelectron microscopy utilizes immunogold labeling of endogenous proteins with specific antibodies to detect and localize these proteins in cells and tissues. The procedure provides insights into the distribution and quantification of protein under different stimulation conditions offering clues about their functions. Conjugating highly electron-dense gold particles with primary or secondary antibodies allow antigen detection on and within cells, with high resolution and specificity.
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Types of Coprecipitation01:10

Types of Coprecipitation

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Coprecipitation is the contamination of a precipitate by otherwise soluble species and occurs via different processes. In colloidal precipitates, coprecipitation occurs via surface adsorption. For instance, barium sulfate has a primary layer of adsorbed barium ions and a secondary layer of nitrate counterions. This results in contamination of the precipitate by barium nitrate.
Sometimes, ions in a crystal lattice can undergo isomorphous replacement by inclusions of similar charge and size. For...
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Immunoprecipitation.

Juan S Bonifacino1, David C Gershlick1, Esteban C Dell'Angelica1

  • 1National Institute of Child Health and Human Development, Bethesda, Maryland.

Current Protocols in Cell Biology
|June 2, 2016
PubMed
Summary
This summary is machine-generated.

This study details selective immunoprecipitation protocols for protein characterization and interaction studies. It includes methods for lysate preparation, antibody binding, and enhanced specificity through antigen reprecipitation.

Keywords:
agarose beadantibodyantigenimmunoprecipitationimmunoprecipitation-recapturemagnetic bead

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Area of Science:

  • Biochemistry
  • Molecular Biology
  • Cell Biology

Background:

  • Selective immunoprecipitation is crucial for protein analysis.
  • Characterizing proteins and their interactions requires robust methodologies.
  • Existing protocols may lack comprehensive steps for diverse cell types.

Purpose of the Study:

  • To provide clear, step-by-step protocols for selective immunoprecipitation.
  • To enable detailed protein characterization and interaction studies.
  • To offer methods for enhancing the specificity of immunoprecipitation.

Main Methods:

  • Detailed protocols for preparing cell and yeast lysates.
  • Methods for antibody immobilization on solid matrices.
  • Procedures for performing selective immunoprecipitation.
  • A technique for antigen reprecipitation to increase specificity.

Main Results:

  • Comprehensive protocols for selective immunoprecipitation are established.
  • Methods cover lysate preparation from various cell types.
  • A reprecipitation technique is presented to improve assay specificity.

Conclusions:

  • The provided protocols facilitate accurate protein characterization.
  • The methods support the study of protein-protein interactions.
  • Enhanced specificity is achievable through antigen reprecipitation.