5ml lysate Bait Proteins Co Immunoprecipitation Protein Protein Interaction

Introduction

Co-IP is a classic technique widely used for protein-protein interaction identification and validation by using target protein-specific antibodies to indirectly capture proteins that are bound to a specific target protein. Co-IP has become an effective method in detecting the interaction between proteins.

The general experimental procedure of co-immunoprecipitation (Co-IP) provided by us is: construct a plasmid of the target gene, co-transform the cells, add an anti-bait protein antibody to the cell lysate, and then add the binding specifically binding to the antibody to Agarose after incubation. Protein A or G on the beads, if there is a protein of interest in the cell that binds to the protein of interest, a complex can be formed: "target protein + bait protein + anti-bait protein antibody + Protein A or G", denatured poly Acrylamide gel electrophoresis, the complex is separated again. The protein of interest is then detected by immunoblotting or mass spectrometry. The target protein obtained by this method is naturally combined with the protein of interest in the cell, and conforms to the actual situation in the body, and the obtained result has high credibility.

CELLFREE provides the Co-IP testing service, including verifying the interaction between known proteins and pulling the unknown protein out of the lysate. The verification scope covers Protein-protein/peptide interactions, antigen and antibody fragment characterization, rizatprotein-small molecule interactions, protein-DNA/RNA interactions, etc.

Service Highlights

• The sensitivity of silver staining technique is 100 times high that of coomassie brilliant blue (CBB) and the SDS-PAGE electrophoresis results of that are more distinct.
• Complete testing equipment, such as liquid mass spectrometry and Fortebio, is furnished as a companion to further analyze the testing results.
• The experiment is repeated twice to eliminate the influence of random factors and make the result more reliable.
• The preparation platform for protein, antibody and so on are facilitated for completing all experiments as long as your protein sequence is provided.

Customer offer

1,The expression plasmid of the bait protein and the target protein needs to be verified by sequencing, and a sequencing peak map file is provided.
2,If it is not necessary to transfect or transform, it is necessary to provide more than 5ml of lysate containing bait protein and target protein. If it is prokaryotic expression, it is required that the protein be expressed in the supernatant.
3,For specific antibody detection using bait proteins and target proteins, at least 20 ul of specific antibodies are required.

We provide

• Bait proteins: protein/sample is provided by client or prepared by us (your protein sequence is required only)
• Prey proteins: protein/sample is provided by client or prepared by us (your protein sequence is required only)
  (Protein sample includes animal and plant tissues, organ samples or their lysate)
• Antibody: specific antibody to bait protein is provided by client

Deliverables

Experimental report: interaction analysis report, quality inspection report, experimental process, SDS-PAGE, silver staining map, etc.