|Product name||Universal IP/Co-IP Toolkit (Agarose)|
|Applications notes||Based on the pain points of the traditional IP/Co-IP experiment, Abbkine developed a Universal IP/Co-IP Toolkit (Agarose); After designed and researched, the toolkit contains: optimized natural and denatured lysate, protein A/G Agarose, IP negative control, IPkineTM second antibody, it can meet the IP/Co-IP needs of most users|
|Kit components||• Non-Denaturing Lysis Buffer-25 mL
• Denaturing Lysis Buffer-25 mL
• 20×Wash Buffer-20 mL
• Protein A/G Agarose-0.45 mL
• Elution Buffer-2 mL
• Neutralization Buffer-0.2 mL
• 100×Proteinase Inhibitor Cocktail-0.2 mL
• Mouse IgG (1mg/mL)-30 μL
• Rabbit IgG (1mg/mL)-30 μL
• IPKine™ HRP, Goat Anti-Mouse IgG LCS-30 μL
• IPKine™ HRP, Mouse Anti-Rabbit IgG LCS-30 μL
|Features & Benefits||• Efficient: Efficient antibody binding capacity and low protein non-specific adsorption rate, saving antibody consumption.
• Convenient and universal: Contain all the necessary buffers needed for IP/Co-IP and WB experiments, it can meet the requirements of sample IP/Co-IP or WB at the same time.
• Reliable and stable: It contains ready-to-use IP negative control, which can eliminate the non-specific binding of IgG itself to the target protein or other specific biomolecules to ensure the specificity of IP antibodies, and contains unique IPkine™ secondary antibodies to perfectly eliminate heavy chain interference.
|Storage instructions||Refer to list of materials supplied for storage conditions of individual components.|
|Precautions||The product listed herein is for research use only and is not intended for use in human or clinical diagnosis. Suggested applications of our products are not recommendations to use our products in violation of any patent or as a license. We cannot be responsible for patent infringements or other violations that may occur with the use of this product.|
|Background||Immunoprecipitation (Immunoprecipitation, IP) is a small affinity purification method using specific antibodies fixed on solid-phase supports such as magnetic beads or agarose. As an important part of many proteomics-related studies, IP can be used to detect the existence of proteins, relative abundance, up-and-down expression of proteins, stability and interaction of proteins and so on.|
Fig.1. Protein was extracted from the non-denaturing Lysis Buffer, then it was verified by Co-IP. While the whole cell lysates (Input) and Co-IP samples were validated with Stat1 monoclonal antibody, Stat2 polyclonal antibody, and GAPDH monoclonal antibody respectively by WB.
The actual volume of Protein A/G agarose microspheres was 450 μL, and the total suspension was 900 μL.
In an IP validation experiment, the Input samples were directly used for SDS-PAGE and WB experiments and could be treated with denatured lysis buffer. The samples used in the IP/ CO-IP experimental operation be treated with non-denatured lysis buffer.
Rule out the possibility of non-specific binding, such as testing a rabbit cell for a protein, if there is no IgG control and the operation has a non-specific protein that interacts with the designed antibody, the result will be positive. If lgG negative control is done, the control does not appear positive, indicating that there is no non-specific protein, the control appears positive, indicating that the antibody has the possibility of non-specific binding to the hybrid protein, need to start the experiment again.
Author：Guo, Haoran, et al. Publication name：Journal of Cellular Physiology IF：5.6
1.The species of antibody reactivity should be the sample species that can be matched normally after Abbkine R&D experts have passed strict scientific verification. If your sample is not within the range of reactivity, in order to improve the efficiency and results of your experiment, it is not suggested to try other species. Otherwise, it may lead to sample mismatch and affect the effect of your experiment.
2.Please aliquot the antibody received as soon as possible and store it at -20℃, avoid repeated freezing and thawing, and use it within one year.
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