Application of phosphatidylinositol kinase (antibody)_Industrial additives

Background^[1-3]

Phosphatidylinositol kinase (antibody) is a type of polyclonal antibody that can specifically bind to phosphatidylinositol kinase. It is mainly used for Western Blot, IHC-P, IF, ELISA, and Co. to detect phosphatidylinositol kinase. -IP and other immunological experiments.

Detection principle: Double-antibody sandwich method to determine the level of phosphatidylinositol kinase in the specimen. Coat the microwell plate with the purified phosphatidylinositol kinase antibody to make a solid-phase antibody. Add phosphatidylinositol kinase to the microwells coated with the monoclonal antibody in sequence, and then combine with the HRP-labeled phosphatidylinositol kinase antibody. An antibody-antigen-enzyme-labeled antibody complex is formed. After thorough washing, the substrate TMB is added for color development. TMB is converted into blue under the catalysis of HRP enzyme and into the final yellow under the action of acid. The depth of color is positively correlated with the phosphatidylinositol kinase in the sample. Use a microplate reader to measure the absorbance (OD value) at a wavelength of 450 nm, and calculate the concentration of phosphatidylinositol kinase in the sample through the standard curve.

Phosphatidylinositol kinase is the collective name for phosphatidylinositol 3-kinase, phosphatidylinositol 4-kinase and phosphatidylinositol 4-phosphate 5-kinase. Enzymes that specifically catalyze the phosphorylation of the 3, 4 or 5 hydroxyl groups on the 1-phosphatidyl-1D-inositol ring, respectively.

Phosphatidylinositol is mainly composed of two parts, one is 1,2-diglycerol phosphate, and the other is inositol. In cells, it plays a very important role in cell morphology, metabolic regulation, signal transduction and various physiological functions of cells.

Apply^[4][5]

For research on the role of phosphatidylinositol 3-kinase/protein kinase B pathway in the pathogenesis of middle ear cholesteatoma

To explore the expression and significance of P-EGFR, P-Akt, cyclinD1 and PCNA proteins in middle ear cholesteatoma epithelial tissue.

Methods: Immunohistochemistry was used to detect the expression levels of P-EGFR, P-Akt, cyclinD1 and PCNA proteins in 40 cases of acquired middle ear cholesteatoma tissues and 20 cases of normal external auditory canal skin tissues, and their differences were analyzed. Ask about the relevance.

Results: The positive rate of P-EGFR protein in 40 cases of middle ear cholesteatoma epithelial tissue specimens was 65.0% (26/40); the positive rate of P-Akt protein was 72.5% (29/40); the positive rate of cyclinD1 protein The positive rate of PCNA protein was 62.5% (25/40); the positive rate of PCNA protein was 80.0% (32/40).

The positive rate of P-EGFR protein in the normal external auditory canal skin tissue of 20 cases in the control group was 20.0% (4/20); the positive rate of P-Akt protein was 35.0% (7/20); the positive rate of cyclinD1 protein was 10.0% ( 2/20); the positive rate of PCNA protein was 45.0% (9/20). After statistical analysis: the expression differences of the four proteins P-EGFR, P-Akt, cyclinD1 and PCNA in the middle ear cholesteatoma group and the normal external auditory canal skin group were statistically significant (P<0.05); middle ear The expression of P-EGFR and P-Akt, the expression of P-Akt and cyclinD1, and the expression of cyclinD1 and PCNA in the cholesteatoma group were significantly positively correlated (P<0.05).

Conclusion: 1. The expression of P-EGFR, P-Akt, cyclinDl and PCNA protein is increased in middle ear cholesteatoma epithelial tissue, and the expression of P-EGFR and P-Akt in the middle ear cholesteatoma group, The expression of P-Akt and cyclinD1, and the expression of cyclinD1 and PCNA were significantly positively correlated, suggesting that the activation of EGFR in middle ear cholesteatoma epithelial cells is accompanied by the activation of its downstream effectors P-Akt, cyclinD1 and PCNA.

2. The EGFR/PI3K/Akt signaling pathway may play an important role in the excessive proliferation mechanism of middle ear cholesteatoma epithelial cells and participate in the pathogenesis of middle ear cholesteatoma.