Application of Flag Tag Antibody in Immunoprecipitation IP & Immunofluorescence IF

The Flag tag system is a well-recognized expression system for expression, purification, and detection of fusion proteins. It is widely used in Western blotting, immunocytochemistry, co-immunoprecipitation, flow cytometry, protein purification, protein interaction, and protein separation. And many other research areas. The Flag tag is a polypeptide fragment consisting of 8 hydrophilic amino acids (DYKDDDDK) fused to a recombinant protein. The Flag tag consists of only 8 amino acids, so it does not occupy other epitopes and binding domains, avoiding changes in the function, secretion, and transport of the fusion protein. Due to its hydrophilic nature, the Flag tag tends to localize to the surface of the fusion protein and is therefore more susceptible to binding to antibodies and enzymatic hydrolysis by enterokinase.

How to choose the appropriate Flag tag antibody?

When choosing Flag antibodies, we mainly need according to our own needs, especially for experimental applications. In addition, the more types of applications after an antibody is detected, the greater the choice in use; while the monoclonal antibodies from mouse sources generally have better specificity, stability, and potency than polyclonal antibodies. In addition, the titer of the antibody is also an important aspect to examine the cost performance of the antibody, which is equivalent to the dilution ratio in practical use. A high titer of 100μl Flag antibody (if its WB assay has a dilution ratio of 1:5000 and the final use solution is equivalent to 500ml), which is 1ml of Flag antibody lower than the potency (if the dilution ratio of WB assay is 1:200) The final use solution is equivalent to 2ml).

The titer of Abbkine's Flag tag antibody was somewhat unexpected. When doing WB experiments, the first recommended 1:1000 according to the instructions, the strips shown are very strong, and then the conditions of continuous dilution and groping, the WB strips at 1:10000 dilution is still very good. ——One client of the Medical Department of Sun Yat - Sen University

Why recommend the Flag antibody from the Abbkine brand?

The Flag system is a system owned by Sigma-Aldrich ^® , so Sigma's Flag tag antibody is the choice of many customers. Sigma's Flag tag antibody mainly has two clones of M1 and M2, of which the Flag antibody of M1 clone can be used to detect the recombinant protein whose Flag tag is located at the N-terminus, while the Flag monoclonal antibody of M2 clone can detect almost all Flag tag protein. Its 2010 China offers are ¥4649/200μg and ¥3683/200μg respectively.

If you want to choose a more cost-effective Flag tag antibody, then Abbkine may be another priority, and its price is about 30% lower than Sigma. Abbkine's Flag tag antibody Anti-Flag Tag Monoclonal Antibody (Catalog #A02010) can be used for the detection and expression of Flag tag fusion protein expression, intracellular localization, and purification, qualitative or quantitative detection of Flag fusion protein (including N-terminal, C End) and so on. The Flag antibody is a mouse-derived monoclonal antibody whose immunogen is a KLH-conjugated Flag tag (DYKDDDDK) polypeptide. In addition, Abbkine also provides HRP-conjugated Flag-tagged antibodies, especially for IP-WB verification to remove secondary antibody interference; rabbit-derived polyclonal Flag antibody also provides customers with another Flag label detection protocol.

Abbkine's Flag monoclonal antibody can be used not only for IF experiments in intracellular localization, but also for IP experiments. The 1:200 or higher IP dilution ratio also indicates that the antibody itself has a high potency, and because it is a monoclonal antibody, its specificity is also high. The high dilution ratio corresponding to the high price also means high cost performance. For the WB experiment, the recommended initial dilution ratio for the specification is 1:1000, but it also performs well at a dilution ratio of 1:5,000 (Abbkine also performed 10,000 dilutions later, still showing a good band display , not listed in the manual). The figure below shows the application of Abbkine's Flag antibody.

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Click image to see more details of Flag tag antibody

A high titer antibody not only affects the sensitivity of the experimental results, but also means a higher price/performance ratio. Abbkine's Flag tag antibody (Cat. #A02010) In the conventional WB experiment, the recommended initial dilution ratio is 1:1000. Actually in Abbkine application, there is a dilution ratio of 1:10,000 or higher, depending on the customer. Specific experimental conditions are optimized.

Abbkine also offers a large package of 1ml Flag tag antibody, which is ideal for the experimental needs of the immunoassay of His tag system. For the convenience of customers, Abbkine is available in stock in China. Abbkine provides global product quality tracking and after-sales service and protection for the Chinese market. If you have any problems during the use, please contact your local agent Ai Meijie Technology Co., Ltd. directly.

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