Dialogue between laboratory experts and experimental rookies

Experimental rookies: When I first entered the laboratory, it was completely a rookie. The boss first asked the literature to read the information, which was a bit confusing. Moreover, there are so many reagents in the literature, as well as recombinant proteins such as cytokines. The difference?

Laboratory experts: Recombinant protein is simply a protein obtained using DNA recombination technology; cytokines are small protein molecules secreted by cells that can induce various physiological reactions such as cell proliferation, growth or differentiation. In fact, they are also obtained using recombinant methods, which are recombinant Part of the protein. However, cytokines include many types, such as immune system regulators such as interleukins and interferons, growth factors such as fibroblast growth factor FGF, vascular endothelial growth factor VEGF, and chemokines such as CCL2 and CXCL8 (chemokine-induced cells Migration or chemotaxis).

 

Experimental rookies: Oh, that’s what it is. Learn today. But I just checked a factor I will use online, there are so many products on the market, how to choose the most suitable product from it, my choice difficulty syndrome is guilty, what should I do? What are the sources of protein products?

Laboratory experts: The most commonly used protein expression systems are mammalian cell expression system and E. coli expression system, in addition to yeast expression system and insect expression system. Some proteins require post-translational modifications to be biologically active. Mammalian cells can provide such modifications, but bacterial cells often cannot. Therefore, when purchasing this type of protein, you should pay special attention to the protein source in the product manual.

Experimental rookies: How do I choose these four expression systems? I suddenly felt a little lack of knowledge, and you need to be able to give pointers!

Laboratory experts: Do not dare to give pointers, but everyone can learn together. Before choosing an expression system, we need to fully understand our research objectives and select the most suitable expression system according to our application. The following table can be used as a reference.

Experimental rookies: I think the cytokines bought by the sister next door are powdered, so strange!

Laboratory experts: The recombinant protein products on the market are basically in the form of lyophilized powder. Only a small amount of the finished product is liquid, and the lyophilized powder is easier to transport and store.

Experimental rookies: How to determine the concentration of added cytokines during the experiment?

Laboratory experts: Since each manufacturer has different production processes or live measurement methods, the activity of cytokines is also different, so the recommended concentration of cytokines can refer to relevant literature or do concentration gradient experiments to determine the best concentration for your own experiments .

Experimental rookies: The molecular weight shown on the gel charts on many cytokine websites is very different from the theoretical molecular weight. What is the reason?

Laboratory experts: First of all, we need to understand two concepts: theoretical molecular weight and apparent molecular weight. The theoretical molecular weight is actually calculated based on the amino acid sequence of the protein; the apparent molecular weight is the protein molecular weight estimated by SDS polyacrylamide gel electrophoresis using a standard reference material of known molecular weight. We all know that the difference between the theoretical molecular weight and the apparent molecular weight is very common in the experiment. The molecular weight of the two on many websites is very different. A large part of it is due to the structure of the protein itself such as protein glycosylation.

Experimental rookies: How to choose the cytokine with good quality?

Laboratory experts: The influence of cytokines on the experiment cannot be underestimated. Once the selection fails, the experiment will not be carried out smoothly, wasting time and effort, so it is very important to choose a cytokine supplier with guaranteed quality. When we choose suppliers, we generally choose companies with good reputation, because companies with good reputation will carry out various quality control on their protein products to ensure the quality of the protein and the experimental effect. It is recommended to read the instructions carefully before purchasing, on the one hand, you can understand the basic information of the protein, on the other hand, the instruction manual will also provide a variety of data such as protein purity, endotoxin content, SDS-PAGE gel map. Of course, biological activity is also a factor to consider when choosing protein products. For example, BaF3 mouse pro-B cells transfected with human CXCR3 were treated with chemokine CCL10 to measure the chemotactic ability of CCL10; TGF-beta 1 was used to inhibit IL-4 induced HT-2 cell proliferation To detect the biological activity of TGF-beta 1.

Product NO Product Name
PRP1012 Human IL-6 protein
PRP1013 Human TNF-alpha protein
PRP1010 Human bFGF protein, His Tag
PRP1011 Mouse bFGF protein, His Tag
PRP1014 Human IFN-gamma protein, His Tag
PRP1015 Mouse IFN-gamma protein, His Tag

Abbkine’s cytokines have high quality guarantee:

  1. Cell-based biological activity assay.
  2. Complete quality control system.
  3. Low endotoxin levels.
  4. High purity,> 95%.
  5. Lyophilized powder form.

Abbkine specializes in the fields of proteinology and cytology, and is committed to innovating and developing various antibodies, proteins, analytical reagents and kits, with a view to becoming a key promoter in the fields of life science research and development, drug research and development. We provide you with the favorite products of protein and immunological research users, from basic immunological products, such as protein extraction and quantification, to the internal reference labeling antibodies, primary antibodies and secondary antibodies of immunological experiments, etc .; the favorite products of cell research users, from Dyes and kits for detecting the state of cells, kits for extracting organelles, staining and tracking of cell substructures and cell metabolism detection products, and cytokine and protein detection kits for cell culture, only to help your research career !

About Abbkine

Our position: Serve global users of cell and protein research, and provide users with economical and technical product solutions through application processes and product portfolios.

Our mission: to stimulate our inner creativity, provide competitive biomedical products and services, and continue to create maximum value for customers.

Our vision: to be a respected, world-class supplier of biomedical products and services.

 

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