Background[1][2]
Monoclonal antibody technology originated from the in vitro mouse hybridoma technology created by Kohler et al. After years of research, mouse monoclonal antibodies have developed from simple biological laboratory research and clinical diagnosis to therapeutic antibody drugs. However, the main drawback of mouse monoclonal antibodies is that the mouse immune system cannot recognize certain immunogens, especially murine-derived immunogens, and the affinity of mouse monoclonal antibodies is not as high as that of rabbit-derived antibodies. However, due to the lack of rabbit-derived plasmacytoma and the technical difficulties in transfecting rabbit B cells with viruses in vitro, it was not until 1995 that a rabbit plasmacytoma cell line was successfully obtained from c-myc/v-abl transgenic rabbits, and the Stable rabbit-rabbit hybridomas led to a breakthrough in rabbit monoclonal antibody technology.
Rabbit monoclonal antibody technology has been improving day by day in recent years, and there are currently thousands of commercialized rabbit monoclonal antibodies. As a unique member of the apoptosis inhibitory protein family, survivin participates in the regulation of cell division and proliferation processes and plays an important role in the inhibition of apoptosis. In view of the characteristics of tissue distribution and relative correlation with most tumors, its research provides strong support for clinical diagnosis and treatment of tumors.
Heat shock protein (HSP) is a type of protein produced under the induction of various stress factors, especially high-temperature environmental factors. It is widely present in the cells of humans, animals, microorganisms and plants and can enhance The ability of cells to resist various damaging factors and promote cell self-repair of damage caused by various stimuli has important biological functions and is closely related to the proliferation of tumor cells. HSPs-peptide complexes activate multiple CTL cloning is expected to specifically kill all tumor cells in the same tumor, break immune tolerance, eliminate residual tumor tissue, and overcome difficulties such as antigen variability and cell heterogeneity that are currently difficult to solve in tumors.
According to their different molecular weights, they can be divided into HSP110, HSP90, HSP70, HSP60 and small molecule heat shock protein subfamilies. Among them, small heat shock protein (sHSP) exists in almost all organisms, has a conserved α-crystalline protein domain, and plays different functions in cells.
Heat shock protein 27 (HSP27), also known as heat shock protein B1 (HSPB1), is an important member of the small molecule heat shock protein family. It is commonly expressed in a variety of tissue cells, especially myocardium. It is the most abundant in terminally differentiated cells such as cells and neurons. It can inhibit cell apoptosis through mechanisms such as increasing the redox state of mitochondria, binding to Fas death domain-related proteins Daxx and Cytc, inhibiting caspase cascade activation, and participating in regulating the proliferation of various cells. , signal transduction of differentiation and apoptosis, and protect cells from damage by various stress factors. In recent years, the relationship between HSP27/HSPB1 and various clinical diseases, especially tumors, has received increasing attention.
Phosphorylated HSP27 (pSer82) rabbit monoclonal antibody can be used in immunohistochemistry techniques such as immunofluorescence, immunoenzyme, immunoferritin, immunogold, and radioimmunoassay. The principle is that the binding between antibodies and antigens is highly specific, and immunohistochemistry takes advantage of this principle. First, a certain chemical substance in tissues or cells is extracted and used as an antigen or hapten. Specific antibodies are obtained by immunizing animals, and then the antibodies are used to detect similar antigenic substances in tissues or cells.
The complex of rubber additives and antigens and antibodies is colorless, so it is necessary to use histochemical methods to display the binding sites of antigens and antibodies in order to characterize unknown antigens in tissues or cells. , positioning or quantitative research. The phosphorylated HSP27 (pSer82) rabbit monoclonal antibody is stored at -20℃, and the Western primary antibody dilution is stored at -20℃ or 4℃. It is valid for one year. It is recommended to store the Western primary antibody dilution at 4°C. If not used for a long time, storage at -20°C may be considered. However, freezing and thawing may cause slight turbidity and a small amount of insoluble matter. If the phosphorylated HSP27 (pSer82) rabbit monoclonal antibody is used in Western blot (WB), immunofluorescence (IF), immunocytochemistry (ICC) and other experiments, please pay attention to recycling the used diluted antibody.
Recovered phosphorylated HSP27 (pSer82) rabbit monoclonal antibody can usually be reused at least 5-10 times. Please store diluted antibodies at 4°C, including diluted antibodies that have been used. Antibodies that are reused after recovery should be used in the same way as freshly diluted antibodies. If you find that the antibody is slightly turbid during repeated use, you can centrifuge it at 10,000g for 1-3 minutes and take the supernatant for subsequent testing. If the recovered antibody shows obvious floc or mold growth, you can consider discarding the antibody.
Main reference materials
[1] Li Tingting, Cui Huifei. Development and application prospects of rabbit monoclonal antibodies [J]. Food and Drugs, 2012, 14(9): 373-376.
[2] Tian Fangyan, Cao Hui, Zhang Peixiang, Free Silica, etc. Preparation and identification of mouse HSP27/HSPB1 monoclonal antibodies [J]. Oncology Pharmacy, 2013 (1): 26-29.
