For the traditional molecular cloning technology, the target gene is amplified by PCR. And then the T-A cloning vector is constructed and transferred to competence E.coli. Next, target gene fragment and vector are digested by the same restriction enzyme. After gel-purify, insert fragment and vector are ligated by ligaseWe will spend 2-3 weeks to complete this process.This technique has many drawbacks,as follows:1.The multiple cloning site is make up of many restriction enzyme cleavage sites,which may be not always suitable for the target gene;2.Adding the restriction enzyme cleavage site during the gene cloning by the PCR, which may bring non-specific amplification; 3.If the target gen......

The topic of quantitative determination of Caspase-1 expression in ATP and LPS stimulated monocytes by ELISA analysis had been introduced. Now we briefly introduce the Caspase family members. The study on Caspase family in cellular apoptosis is very popular in recent years. 14 Caspase members have been cloned, and they are similar with amino acid sequences and structure of proteins. Caspase protein usually exists in forms of inactive zymogen. Caspase members are divided into 3 classes as for their different function and sequence in cascade reaction. The first members of the Caspase family include Caspase- 2, Caspase- 8, Caspase- 9 and Caspase- 10. They are initiators of apoptosis, and......

Internal control is a reference in the experiment system, regularly, internal controls in mammalian cell expression system are proteins expressed by house keeping genes. These proteins show relatively constant quantity in different tissue and cells. So, when Western Blot is designed for protein level test, the internal control protein could be considered as a reference to further ensure the accuracy of the result. Last time we introduced the three commonly used internal controls including beta-actin, beta-tubulin, GAPDH (glyceraldehyde-3-phosphate dehydrogenase). This time we’ll present other internal controls produced by Cloud-Clone Corp. Sodium/potassium-transporting ATPase (N......

Collagen, mainly found in animal connective tissues, is the most abundant mammal protein, accounting for 25-35% of the total body protein content. Collagen can be divided into fiber type and non-fiber type. The most common types are Type I, Type II, Type III, Type IV, Type V. Wherein Type I, II, III, V are fibrous, Type IV is non-fibrous. So far 28 kinds of collagen are found, while over 90% are Type I. Collagen can be used for cosmetic surgery, bone grafts, tissue regeneration, wound healing, etc. Further, collagen mutations lead to diseases, including osteogenesis imperfecta, achondroplasia, osteoporosis and so on. Consequently, research on collagen has significant meanings, and ha......

Immunohistochemistry can be used for qualitative and semi-quantitative detection, aswell as location of antigen in situ in cells of a tissue section. Immunohistochemistry has the advantage of higher sensitivity, it is one of the most common immunological detection methods. However, due to the complicated procedures, such as antigen retrieval, determination of antibody’s concentration and appropriate staining, it always will lead to unsatisfied results. By popular demand, Cloud-Clone Corp. begins to provide IHC service for customer. IHC protocols Advantages There is a huge storage of semi-finished materials, more than 19,000 kinds of antibodies could be sent out immediately; ......

Antibody labeling is a common technology used in sample preparation for immunoelectron microscopy. Due to the simple operation, high sensitivity, it is widely used in various immunoassays such as IHC, WB, ICC, ELISA. The labels can amplify the detection signal for antigens, making it a fast and economic detection technology in the market. Commonly used molecules for antibody labeling include enzymes, fluorescent molecules, isotope, biotin, colloidal gold etc. Below is the introduction of conjugated antibodies from Cloud-Clone Corp. 1. Horseradish peroxidase (HRP) labeling. HRP is conjugated to monoclonal/polyclonal antibody by periodate method. The glycosyl of HRP is oxidized into al......