Home Technique / Proteomics and protein biochemistry / Immunoprecipitation
| Immunoprecipitation (IP) is the technique of precipitating an antigen out of solution using an antibody specific to that antigen. This process can be used to enrich a given protein to some degree of purity. Co-immunoprecipitation (also known as a pull-down) can identify interacting proteins or protein complexes present in cell extracts: by precipitating one protein believed to be in a complex, additional members of the complex are captured as well and can be identified. The protein complexes, once bound to the specific antibody, are removed from the bulk solution by capture with an antibody-binding protein attached to a solid support. |
General Principles of Immunoprecipitation
new recommended
protocol
This is the Bart's cookbook on immunoprecipitaiton. It gives some general principles on performing immunoprecipitation using different lysis buffers.(Sefton Lab, Salk Institute).
Content:
RIPA buffer
NP40 buffer
PBS
TN
Boiling in SDS
Antisera ...
Anti-FLAG Co-Immunoprecipitation
protocol
Immunoprecipitation is a technique that permits the purification of specific proteins for which a antibody has been raised. This primary antibody is either already bound to agarose or can be bound to the protein A/agarose beads during the procedure in order to ...
CO-IMMUNOPRECIPITATION ANALYSIS OF PROTEIN-PROTEIN INTERACTIONS
protocol
This protocol uses total cell extracts to analyze putative protein-protein interactions in eukaryotic cells. One can also use nuclear extracts as a source for this protocol (Morris lab at University of Pennsylvania). ...
Immunoprecipitation protocol from Sigma
protocol
Immunoprecipitation (IP) is one of the most widely used immunochemical techniques. Immunoprecipitation followed by SDS-PAGE and immunoblotting, is routinely used in a variety of applications: to determine the molecular weights of protein antigens, to study pro ...
Immunoprecipitation and co-immunoprecipitation
new recommended
review
Immunoprecipitation technique was first developed in 1974 to resolve the immunoprecipitated proteins on slab gels [1] and has since then become one of the most important techniques to study protein-protein interactions.
Two major factors affect the success ...
Phosphatase inhibitors cocktails selection guide
new
review
Selection guide for phosphatase inhibitors cocktails (Merck).
Phosphatase inhibitor cocktails is used to prevent dephosphorylation of proteins in protein biochemistry applications. Commonly used protein phosphatase inhibitors protect dephosphorylation of se ...
Protein A Sepharose Regeneration
troubleshooting
I am using Protein A Sepharose (Pharmacia) for immune precipitations. I
elute immune complexes after binding with laemmli buffer (SDS-PAGE
loading buffer).
Is there anyone who has experience, whether this very expensive material
can be regenerated ...
How to remove RNase from immunoprecipitation?
troubleshooting
We are trying to do a "simple" immunoprecipitation in which
the unbound fraction will be assayed for RNA processing activity. We find that
protein A-agarose beads have little non-specific RNase activity. After adding
serum (immune or non-immune), we w ...
Immuno-precipitation of Phospho-Tyr proteins
troubleshooting
I am trying to assess the degree of phosphorylation a receptor
undergoes in response to it's ligand plus various other factors.
If I apply the ligand to my cells and then extract in the presence of
sodium orthovanadate and do a western using an antibo ...
Immunoprecipitation western blot troubleshooting
troubleshooting
In an immunoprecipitation-western blot I used rabbit polyclonal antibody and protein A agarose beads(protein A is covalently bind to beads) to pull down antigen complex. In following western blot I used rabbit polyclonal antibody as primary antibody and HRP-an ...
troubleshooting tips of IP, western and immunostaining
troubleshooting
General troubleshooting tips of immunoprecipitation, western blot and immunohistochemistry from Chemicon.
Link: troubleshooting tips of IP, western and immunostaining
This link was lasted ...
Troubleshooting Tips for Immunoprecipitation / Kinase Assays
new
troubleshooting
A brief troubleshooting guide on immunoprecipitation - kinase assay. (Upstate)
Determine the optimal stimulation time and temperature for each individual kinase that is to be tested. This is essential since optimal stimulation periods are often short and ca ...
Immunoprecipitation TroubleShooting Guide
new
troubleshooting
Troubleshooting guide from Sigma. It addresses immunoprecipitation problems on:
1. No binding
2. High background or unwanted precipitates
...
co-immunoprecipitation question: starting concentration
troubleshooting
What is a good starting concentration of each protein to have? What volume? I know that my proteins are tight-binding, so dissociation should not be a big problem. (post from NWFSC forum). ...
Immunoprecipitation troubleshooting guide
troubleshooting
A Immunoprecipitation troubleshooting guide from Stressgen Biotechnologies. ...
Troubleshooting the immunoprecipitation procedure
new
troubleshooting
A troubleshooting guide on Immunoaffinity purification (Immunoprecipitation) procedure from Roche. ...
Immunoprecipitation troubleshooting
new
troubleshooting
A troubleshooting guide on immunoprecipitation. (Agrisera)
Problems:
No binding
High background signal
...
More immunoprecipitation protocols
(Protein kinase protocol from Methods in Molecular Biology)
Immunoprecipitation and western blotting detection of proteins
Immunoprecipitation techniques and kits
(Forum)
Protein amount for immunoprecipitation
(Forum)
Methods to identify cellular proteins
Last update: 
10-Jun-2013 05:36 am
