Protein purification topic | protein purification tossed for half a month without results, watch the sharing?
At present, there are many purification methods for recombinant proteins produced by genetic engineering technology, which can be divided into precipitation technology, chromatography technology, double-liquid phase extraction technology, etc. No matter which method is used, the principle of separation and purification is to use the difference of its physical and chemical properties. Physical properties include molecular size, shape, solubility, and chemical properties include isoelectric point, hydrophobicity, and affinity with other molecules. nature, etc. The purification of protein is roughly divided into two stages: crude separation stage and fine purification. Crude separation mainly separates the target protein from other cellular components such as RNA, DNA, etc., such as ammonium sulfate precipitation. Fine purification is to distinguish the target protein from other…
Redefining cell migration assays. The Abbkine Cell Migration Assay Kit is online!
If you are still using the cell scratch method for cell migration assays, you are out of date. Cell migration assay is a method to detect the ability of cell migration and repair, which is commonly used to evaluate wound healing, cell differentiation, embryonic development and tumor metastasis. Although the traditionally used cell scratch method to detect cell migration can be visually observed, it cannot quantify cell migration. Abbkine Cell Migration Assay Kit, from a new perspective, integrates the visualization and quantification of cell migration. Principle of Abbkine Cell Migration Assay Kit Cell migration is the movement of cells after receiving a foreign signal or sensing a gradient of certain substances. It usually occurs in processes such as wound…
The indicators you need, we have them all!
Biochemical kits are mainly developed using the principle of chemical reaction/enzymatic reaction or substance structure characteristics to determine substance content or enzymatic activity in a sample. The CheKine™ series of biochemical detection kits independently developed by Abbkine have a complete range of biochemical detection kits with a wide range of applications. The detection analytes include oxidation/antioxidation, glutathione, amino acid metabolism, glycolysis, fatty acid metabolism, oxidative phosphoric acid Chemical, coenzyme I/II, ATP and other series. The types of samples tested involve animal and plant tissues, serum, plasma, cells, bacteria, various body fluids, etc. The main detection instruments are microplate reader and spectrophotometer. Abbkine's CheKine™ series of biochemical detection kits use more efficient and innovative detection technology and optimized experimental protocols. The…
Easy to deal with Immunoprecipitation, "IP new weapon" is coming
1.Background principle: Immunoprecipitation refers to a method of preliminarily separating the target protein by precipitating the antigen (usually the target protein) from the mixed system by using the characteristic that the antibody can specifically bind to the antigen. Immunoprecipitation is developed based on the traditional affinity purification method. 2.Questions: Although the principle is not difficult, the result is not easy. 1. How many researchers are struggling in the sea of searching for the target strips, 2. How many people are foolishly unable to distinguish the strips under the interference of light and heavy chains; 3. How many people are struggling "Lost yourself" in complex manipulation experiments. Solution: Abbkine has launched a series of IP toolboxes to directly address the…
Encyclopedia of biochemical detection indicators
1、 Definition of biochemical detection Biochemical testing refers to the use of biological or chemical methods to test samples. The determination indexes are usually biochemical indexes such as enzymes, carbohydrates, lipids, proteins and non-protein nitrogen, inorganic elements or body function indexes. In scientific research, biochemical detection provides various conveniences for material content determination, enzyme activity rapid detection and drug development. 2、 Classification of biochemical detection indexes a.Oxidation and Antioxidation Series Oxidation is a common chemical reaction in life, free radicals are produced in the process of oxidation, and antioxidation is a molecule that scavenges free radicals in the body. Enzymatic and non-enzymatic antioxidants play a crucial role in protecting against peroxidative damage caused by exercise. Common oxidation and antioxidant series detection indicators mainly include…
Protein Quantification Topic|What protein quantification methods have you used over these years?
Protein quantification is an important step that must be taken before protein isolation and analysis, protein chromatography separation, protein electrophoresis analysis, protein immunosegregation and analysis, total protein quantification of cell split interpretation, labeling of protein molecules, protein structure analysis, etc., all need reliable quantitative analysis as the basis. The fields of application of protein quantification analysis include biological sciences, food testing, clinical testing, and more. Specifically, we need to use protein quantification experiments in the following scenarios: 1. When judging whether the protein is expressed, first lyse the cells to verify whether the cell lysis is thorough, or to carry out the usual experiment or standardized preservation of the sample, then the cell lysate should be quantified for protein; 2.…
Universal IF Toolkit|Arrange the immunofluorescence experiment clearly
When it comes to immunohistochemistry, as the name suggests, immunohistochemistry (IHC) uses antibodies to detect the localization of proteins and other antigens in tissue slices. Antibody-antigen interactions are detected by enzyme substrate colorimetry or by fluorescent dyes. Although IHC is not as quantitative as Western blotting or ELISA, it can provide valuable information on protein localization in the complete tissue background. Protein expression profiling is of great value to pathologists and can also be used as an important diagnostic tool. The key to the success of IHC experiments lies in a stable, optimized, and reproducible staining protocol that uses high-quality specific reagents. Here we will briefly talk about a major branch of immunohistochemistry immunofluorescence (IF), immunofluorescence experiments seem simple, but…
Beyond BrdU, a new generation of cell proliferation imaging analysis kit (EdU method)
1、 What is EDU? EdU, named as 5-ethynyl-2-deoxyuridine, is a nucleoside analogue of thymine. Its alkyne groups are rare in natural compounds and can replace thymine (T) to infiltrate into the synthetic DNA molecule during the period of DNA replication for DNA synthesis. It is widely used in experimental researches such as cell proliferation. 2、 What is BrdU? The name of BrdU is 5-bromo-2 ' -deoxyuridine. BrdU is a synthetic thymine analogue of thymidine, which is incorporated into the newly synthesized genomic DNA in the mitotic S phase. Subsequently, DNA denaturation was performed to determine BrdU insertion and any other cellular surface and / or intracellular target of interest through cell staining. 3、 Detection principle of Edu EdU can be incorporated into the newly…
Immunofluorescence Master Key - Universal IF Toolkit
We usually encounter many difficult problems when doing immunofluorescence experiments, such as weak staining or no signal, high background and strong non-specific signal, and for example, the hard work of slices, which was discovered only during the final observation. And for example, all the reagents for immunofluorescence are self-prepared, and the configuration time is also real. So how to solve these very common problems? Let's take a look. Immunofluorescence technology (IF) is based on the principle of antigen-antibody reaction. First, a known antigen or antibody is labeled with fluorescein to make a fluorescent antibody, and then the fluorescent antibody (or antigen) is used as a probe to detect tissue or cells. the corresponding antigen (or antibody). Based on the tedious…
Distinction between apoptosis, Pyroptosis, necrosis, iron death and autophagy
“To be, or not to be, that is the question”， To live or die, cells have to choose not only whether to die, but also how to die. There are many "patterns" of cell death, such as apoptosis, Pyroptosis, Ferroptosis and so on. Today, let's talk about cell death. What is apoptosis? Apoptosis is a fundamental biological phenomenon of cells that plays an essential role in the removal of unwanted or abnormal cells in multicellular organisms. It plays an important role in the evolution of organisms, the stability of the internal environment and the development of multiple systems. Apoptosis is not only a special type of cell death, but also has important biological significance and complex molecular biological mechanisms. Apoptosis is a…
