Background[1-3]
Glucose-6 phosphate dehydrogenase antibody is a type of polyclonal antibody that can specifically bind to glucose-6 phosphate dehydrogenase. It is mainly used in experiments related to the detection of glucose-6 phosphate dehydrogenase in vitro.
Glucose 6-phosphate dehydrogenase is the first enzyme EC1 in the phosphogluconate pathway (pentose phosphate pathway), another glucose decomposition pathway other than the glycolysis pathway and the citric acid cycle. 1.1.49.
It has long been known as 6-phosphoglucose dehydrogenase (zwischenferment), which catalyzes the dehydrogenation of 6-phosphoglucose to form 6-phosphogluconic acid. Use NADP as the electron acceptor. The balance of the entire reaction tends toward the production of NADPH, which is used in reductive biosynthetic reactions such as fatty acids. This enzyme exists in the soluble fraction of cells.
Glucose-6-phosphate dehydrogenase
Glucose-6-phosphate dehydrogenase is mainly involved in the pentose phosphate pathway and can also slowly act on other sugars such as β-D-glucose. This enzyme can be activated by its substrate glucose-6-phosphate. If the human gene G6PD expressing glucose-6-phosphate dehydrogenase is defective, it will lead to glucose-6-phosphate dehydrogenase deficiency and cause non-immune hemolysis. sexual anemia. Glucose-6-phosphate dehydrogenase exists in many biological cells. Higher plants have several protein isoforms of glucose-6-phosphate dehydrogenase, which are distributed in the cytoplasmic matrix and peroxisomes. and other cellular structures.
Apply[4][5]
Establishment and application of a method for localization and quantification of glucose-6-phosphate dehydrogenase in sea urchin (Ps ammechinus miliaris L.) fertilized eggs
Using the model organism sea urchin as the test material, using low-temperature freezing semi-thin sectioning technology, two methods, immunogold and silver staining and immunofluorescence, were studied and compared, and their respective characteristics and applicability were summarized in the experiment;
Using these two technical methods, we conducted localization and quantitative research on the key enzyme protein in the fertilization process of sea urchin egg cells-glucose-6-phosphate dehydrogenase;
In addition, using in vivo image analysis technology and “grayscale-absorbance” technology, using glucose-6-phosphate dehydrogenase as the target protein, the feasibility of real-time measurement of enzyme protein activity changes during the fertilization process of living egg cells was explored. And the activity changes of glucose-6-phosphate dehydrogenase during fertilization were analyzed.
(1) Using immunogold and silver staining technology based on low-temperature frozen semi-thin sections, the labeling is highly durable. This method is especially suitable for quantitative analysis of enzyme proteins and in combination with other histochemical methods. Activity research; and immunofluorescence technology, clear activity labels can be easily observed, which is a suitable means to study enzyme protein localization.
(2) Based on the comparative analysis of these two technologies, the subsequent experiment used immunofluorescence technology to perform a localization analysis of glucose-6-phosphate dehydrogenase during the fertilization process of sea urchin egg cells. Studies have found that in unfertilized egg cells, glucose-6-phosphate dehydrogenase is mainly accumulated in cortical granules or attached to the endoplasmic reticulum membrane. After fertilization stimulation, glucose-6-phosphate dehydrogenase is released from the endoplasmic reticulum and cortical granules. At the same time, the immunogold-silver staining method was used to quantitatively analyze the changes in the protein amount of glucose-6-phosphate dehydrogenase during the fertilization process of sea urchin egg cells. The measurement results showed a gradual downward trend. Single-factor variance and homogeneous subset analysis showed that there was a significant difference between the G6PDH content in unfertilized egg cells and the G6PDH content in fertilized egg cells (p<0.05).
(3) The activity of glucose-6-phosphate dehydrogenase during fertilization was studied using in vivo image analysis technology. Through the organic combination of biochemistry, physical optics, and imaging, the experiment achieved real-time monitoring and control of the activity of glucose-6-phosphate dehydrogenase during the fertilization process of surviving sea urchin eggs without damaging the cells. Quantitative analysis.
