How do you select cytokines, How do you use them?

Stimulate the cells cultured in vitro, or as the positive control in ELISA test, we need to use cytokines, often need to use in scientific research and purification of cytokine, so choose the right cell factor is a top priority.

So how do you pick the cytokines and how do you use them?

Don't panic. Here are the books for you.

A:Just entered laboratory, completely is a small white, the boss just let look up the literature data, a little scratching their heads, and so much reagent is mentioned in the literature, still have the recombinant proteins such as cytokines, recombinant proteins and cytokines have what differentiation?

B:Recombinant protein is simply a protein obtained from recombinant DNA;

Cytokines secreted by the cell, or to induce cell proliferation, differentiation and other physiological reaction of protein molecules, is also using the method of restructuring, is part of the recombinant protein.

However, cytokines including many types, such as immune system regulation, such as interleukin, interferon FGF fibroblast growth factor, vascular endothelial growth factor VEGF and other growth factors, and chemokine CCL2, CXCL8 (chemotactic factor inducing cell migration or chemotaxis).

A:Oh oh, that's the way it is. I learned it today.

But I've just checked on the net a I'm going to use factor, unexpectedly there are so many of the same products on the market, how to choose the most suitable products, and made my choice difficulty syndrome, how to do?

What are the sources of protein products?

B:The protein expression system is most commonly used in the expression system of the mammalian cell and the expression system of e. coli, as well as the yeast expression system and insect expression system.

Some proteins need to be translated and modified to have biological activity. Mammalian cells can provide such modification, but bacterial cells often cannot.

Therefore, you should pay special attention to the protein source in the product specification when choosing such proteins.

Next, let's take a look at the characteristics of different presentation systems.

A:How do I choose these four expression systems? I feel a little lack of knowledge at once, and I need you to give directions.

B:I don't know, but you can study together.

We need to fully understand our research purpose before choosing the expression system, and select the most appropriate expression system according to our own application. The following table can be used as a reference.

A:I think the cell factor bought by next door is powder. It's strange

B:The recombinant protein products on the market are basically freeze-dried powder, and only a small amount of finished products are liquid, and freeze-dried powder is easier to transport and preserve.

A:How to determine the concentration of cytokines in the experiment?

B:Because each manufacturer production process or live test method is different, the activity of cytokines are also different, so the use of cytokine concentrations suggest may reference related literature or concentration gradient experiment and used to determine the best use of the suitable experimental concentrations.

A:A lot of cells have a lot of molecular weight and theoretical molecular weight shown in the graph. What's the reason?

B:First, we need to understand two concepts: theoretical molecular weight and apparent molecular weight.

The theoretical molecular weight is actually calculated according to the amino acid sequence of the protein;

The molecular weight of the apparent molecular weight is the molecular weight of the protein by SDS polyacrylamide gel electrophoresis.

We all know that in the experiment theory of molecular weight and apparent molecular weight difference is very common phenomenon, many websites both molecular weight is quite different from above, of which a large part of the reason is that protein glycosylation, etc. The influence of the structure of the protein itself.

A:How to choose the cell factor of quality clearance?

B:Cell factor's influence on the experiment to be reckoned with, once choose failure will directly lead to can't experiments go smoothly, a waste of time and effort, so choose supplier quality assured cellular factors is very important.

When choosing a supplier, we usually choose a reputable company, because reputable companies have various quality control over their protein products to ensure the quality and experimental results of the protein.

It is recommended to read the instructions carefully before the choose and buy, on the one hand, can know the basic information of protein, on the other hand also provide protein in the instructions for purity, endotoxin content, sds-page gel figure such as a variety of data.

Of course, biological activity is also a factor to be considered when choosing protein products.

For example, CXCR3 BaF3 mouse pro - B cells were transfected with chemokine CCL10 to measure CCL10's chemotactic ability.

TGF - beta 1 was used to inhibit the proliferation of il-4 induced ht-2 cells, thus detecting the biological activity of TGF - beta 1.

A:Wow, I have learned a lot of things from chatting with you. Thank you very much for your patience. Thank you for your recommendation. I am going to do the experiment, and I will discuss with you some other time.

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