Three minutes to take you through the problem of immunofluorescence
Immunofluorescence (IF) is a common experiment in immunology experiments. For the experimental masters, it may be the effort of sprinkling water, and a "dazzling and beautiful" film will come out, but for the novice operator, it may be in the overall process. It is not very friendly, and there will still be situations such as fluorescent bright spots in the entire cell sheet, or weak or no signal, too few stained cells, or only the blue light of DAPI in the observation field, and the target fluorescence has almost no or no signal. If so, it might be very frustrating. Today, let's walk into the world of immunofluorescence and see how to sort out our experiments. 1.Principle Immunofluorescence is a…
New products have been added to fatty acid metabolism series
Previously, we provided detailed knowledge about fatty acid metabolism, including fatty acid anabolism and catabolism, and also introduced two biochemical indicators related to fatty acid metabolism, including： 1.fatty acid synthase Fatty acid synthase (FAS) is a multifunctional homodimeric enzyme protein that is the primary enzyme required for the anabolic conversion of dietary carbohydrates to fatty acids. Fatty acid synthase catalyzes the conversion of acetyl coenzyme A and malonyl coenzyme A to long-chain saturated fatty acids in the presence of NADPH. Human fatty acid synthase is a large homodimeric multifunctional enzyme for the synthesis of palmitic acid. The unique carboxy-terminal thioesterase structural domain of fatty acid synthase hydrolyzes the growing fatty acid chains and plays a key role in regulating the…
The strength of IP family has been expanded again, and IP kit of ta protein has returned with great success
Small make up previous published an article called "dry | precipitation utility arranged IP/Co - IP experiment" plainly, we talked about: In the IP/Co-IP experiment, the antibody captures the protein (antigen) and the bead captures the antibody-antigen complex. Now, the IP/ co-IP experiment could eliminate many of the tedious steps involved in the love-making process by "binding" beads to antiboy-antigen complexes. Xiaobian takes you to sort out the steps and attention points of the traditional IP/ co-IP experiment and the optimized and upgraded IP/ co-IP experiment, so that you can remain invincible in the pursuit of love. Based on the above innovative ideas, the research team of Iacoin made unremitting efforts to develop the label Protein label Protein immunoprecipitation kit,…
Analysis and detection of antioxidant capacity
1.What is antioxidant capacity? There are many antioxidants in living organisms, such as SOD, CAT, GSHPx, G6PD, GR, glutathione, etc., which can remove various reactive oxygen species in the body to prevent the generation of oxidative stress induced by reactive oxygen species. The total level of various antioxidant macromolecules, antioxidant small molecules and enzymes in a system reflects the antioxidant capacity of the system. Although the analysis of ROS induced oxidative damage is widely used as an analysis indicator, it is also important to detect the level of antioxidant capacity of cells in vivo, cells and extracts. The antioxidant capacity of intracellular macromolecules can be measured by ORAC (antioxidant capacity of oxygen group), HORAC (antioxidant capacity of hydroxyl group) and…
GAPDH, Tubulin, Actin, how should I choose?
We all know that there are many uncertain factors in the Western blot experiment, so the rigorous design of the Western blot experiment requires a good reference system, such as Marker, positive control, internal reference and so on. Today we will focus on the internal parameters of the WB experiment. 1.What is the internal reference? What are the functions of internal reference? Internal reference is internal reference (Internal Control). For mammalian cells, proteins (Housekeeping Proteins) are generally encoded and expressed by butler genes. Because they are encoded and expressed by butler genes, their expression in various tissues and cells is relatively constant. Therefore, it is commonly used as a reference to detect changes in protein expression levels. The experimental operation…
Loading control topic | Loading control detection，the yardstick for measuring WB experiments
It is well known that the internal reference protein has a high and stable expression in most tissues and cells, which is an important reference to indicate that the samples in each well have the same protein load. Internal reference antibodies can also indicate the efficiency of the transfer membrane during western blotting. Abbkine offers a full set of monoclonal, polyclonal reference antibodies and reference cocktail sets with high specificity and a wide variety of application types to meet the needs of most of your protein qualitative, quantitative and localization analysis. In addition to the common human, mouse and rat antibodies, we also provide internal reference antibodies of more other species, as well as large package customization services. How to…
Immunoprecipitation practical tools magnetic beads and agarose resin performance competition
IP experiments are the choice of proteomics research partners when we need to concentrate proteins in small amounts and perform a single assay. Co-IP is almost mandatory when we need to demonstrate protein-protein interactions. Using cell lysis samples, the interaction of intracellular proteins was proved and the molecular mechanism of the phenomenon was elucidated. At this time, the practical tool for IP experiment is the most urgent problem to be solved by the majority of proteomics researchers. Then how to choose the most suitable IP tool for their primary antibody? Here is a comparison of IP utilities (magnetic beads and agarose resin) : First, because IP was developed as an improved method for column affinity chromatography, it was initially done…
Everything you want to know about protein extraction is here!
1.What is protein extraction? The extraction of protein refers to the process in which the treated or broken cells are put in certain conditions and solution so that the extract can be fully released. 2.Why do protein extraction? Protein extraction is compatible with different downstream experiments, including reporter gene detection, protein purification, immunoblotting, immunoprecipitation, EMSA (also known as gel shift), footprinting, enzyme activity analysis； a.ELISA: The soluble antigen or antibody is bound to solid phase carriers such as polystyrene, and the specific binding of antigen and antibody is used for qualitative and quantitative detection of immune reaction; b.WB: The protein samples were separated according to molecular weight by polyacrylamide electrophoresis, then transferred to the hybrid membrane (blot), and then the…
The convenient method of protein extraction
When we conduct various biological experiments, we often deal with proteins. Proteins are indispensable organic substances for life, and they are also the verification objects of our immunological experiments (WB/IP/IF/ELISA, etc.). Therefore, The beginning of a perfect experiment is protein extraction. How to quickly and completely extract protein? How to efficiently perform protein extraction? This is a problem we often encounter in experiments. A good start is half of the success of the experiment. Let's explore the mystery of the beginning together. 1.Background of protein extraction In the process of protein extraction and preparation, since there are many types of proteins in the sample, the properties are very different. Even if they are the same type of protein, the extraction…
Fat is not guilty, please digest my calories
Fat, a substance we know a lot about but don't know a lot about, began to have a negative social image. The word "fat" is immediately associated with overweight bodies, unhealthy diets and certain chronic diseases. Is fat really so bad? What does it have to do with obesity? It is an important building block of human tissue and one of the main sources of heat (calories). Our vertebrate ancestors apparently took advantage of fat as a super-high-energy fuel, developing unique fat cells and the adipose tissue that lay the foundation for obesity today. In modern times, when food supplies are abundant, people often do not want to store fat and calories, so keeping fit (health) and figure (slim) has…
