IP-specific nano-secondary antibody---efficiently eliminate interference of light and heavy chain

Anti-Mouse IgG for IP, AlpSdAbs^® VHH(HRP) is a new generation of nano-secondary antibody specifically designed for IP, launched by AlpVHHs (Chengdu AlpVHHs Co., Ltd). IP-specific nano-secondary antibodies only recognize conformational IgG, thus avoiding interference of light and heavy chain of antibodies in traditional methods.

Immunoprecipitation (IP) is a commonly used experimental method for purifying and enriching target proteins using antigen-antibody specific reactions. After the antibody binds to the target protein, it is incubated with protein A/G-coupled agarose or sepharose beads to isolate the bead-protein A/G-antibody-target protein complex. The target protein and other proteins or protein complexes that interact with it can then be detected by WB.

However, in immunoprecipitation and co-immunoprecipitation (co-IP) experiments, limitations in primary antibodies often necessitate the use of the same primary antibody or a primary antibody from the same species for both IP and WB. In such cases, using a conventional anti-species IgG enzyme-linked secondary antibody will result in WB detecting the heavy (approximately 55 kDa) and light (approximately 25 kDa) chain produced by denaturation of the IP antibody. Because the amount of antibody used in IP is often significantly greater than that used in WB, nonspecific signals can partially or even completely obscure the signal of the target protein, particularly bands near the heavy and light chains, significantly impacting the accuracy of experimental results.

After a long period of exploration, researchers have summarized the following three experiences to help you avoid interference from light and heavy chain during IP detection:

Case 1:

For WB detection, select primary antibodies of different species from those of IP to avoid the influence of IP primary antibodies.

By using antibodies from different species, the influence of heavy and light chain signals can be effectively avoided during WB detection. However, if there are no multiple species sources of primary antibodies available on the market, it will greatly restrict the conduct of experiments. At the same time, the need for different species sources of the same type of antibodies greatly increases the project's expenditure cost.

Case 2:

For WB detection, select a secondary antibody that reacts only with the heavy or light chain.

To avoid the influence of light and heavy chain, choose a secondary antibody that reacts only with heavy or light chains based on the molecular weight of the target protein during WB. If the molecular weight of the target protein is close to 25kD, choose a heavy chain-specific secondary antibody to avoid interference from the antibody light chain. If the molecular weight of the target protein is close to 55kD, choose a light chain-specific secondary antibody to avoid interference from the antibody heavy chain.

To avoid the complexity and hassle of selecting the right antibody during the actual operation, AlpVHHs now provides a simpler and more effective solution, launching a dedicated secondary antibody for post-IP that specifically recognizes mouse or rabbit light chain or heavy chain, so that the influence of light and heavy chains will no longer be an obstacle to IP experiments.

AlpVHHs provides the following IP-specific nano-secondary antibody products:

Case 3:

Conformational recognition secondary antibody

Anti-Mouse IgG for IP, AlpSdAbs^® VHH (HRP) - IP-specific nano-secondary antibody is a new generation of IP-specific nano-secondary antibodies launched by AlpVHHs. This IP-specific nano-secondary antibody detects primary antibodies in their native conformation and cannot recognize the denatured heavy and light chain of the IP antibody. This effectively avoids interference from the heavy and light chain, ensuring more accurate IP assays. Furthermore, this IP-specific nano-secondary antibody retains the high affinity, strong binding capacity, and extreme stability of nanoantibodies, enabling efficient, rapid, and stable binding to target proteins. Recombinantly expressed, this IP-specific nano-secondary antibody offers consistent product quality, significantly reducing batch-to-batch variability and making experiments more stable and simple, bringing peace of mind to your IP experiments.

AlpVHHs is a leading nanoantibody research and development, production, and service provider in China. AlpVHHs is committed to developing and launching more efficient, stable, and simpler antibody reagents, making scientific research faster, more accurate, and more reliable. For more details, please visit our website https://www.alpvhhs.com.

AlpVHHs provides the following IP-specific nano-secondary antibody product: