Novel serology biomarker GSK3beta detection method for cognitive impairment of diabetic
A technology for diabetes and patients, applied in the field of detection of a new serological marker-GSK3β, which can solve the problems of unsatisfactory specificity and sensitivity, and achieve the effect of simple operation
- Summary
- Abstract
- Description
- Claims
- Application Information
AI Technical Summary
Problems solved by technology
Method used
Image
Examples
Embodiment 1
[0040] Example 1: Dot-blot method to detect platelet GSK-3β activity in diabetic patients
[0041] 1. Platelet separation and purification.
[0042] 1.1 Human whole blood 5ml, 60g, centrifuged at 4 degrees Celsius for 15 minutes, the upper layer is plasma, the middle layer is white blood cells, and the lower layer is red blood cells. Plasma, 60g, centrifuged at 4°C for 15min, to remove sediment, centrifuged at 1500g for 15min, divided into 2 layers, the upper layer was serum, and the lower layer was platelets.
[0043] 1.2 Platelet samples were washed with modified Tyrode's solution, centrifuged at 1500g for 5min three times, and stored in a -80°C refrigerator.
[0044] 2. Platelet protein extraction and concentration determination.
[0045] 2.1 Add pre-cooled lysate 120μl 2×Buffer to platelets, said 2×Buffer consists of 50mM Tris-Cl pH 8.0, 1% NP-40, 150mM NaCl, 0.1% SDS, 0.02% NaNR3R, 20% glycerol, cocktail, Composed of 1% PMSF, after standing on ice for 20 minutes, the p...
Embodiment 2
[0054] Example 2: Enzyme activity kit detects platelet GSK-3β activity in diabetic patients
[0055] 1. Preparation of samples to be tested
[0056] 1.1 Prepare the purified platelet sample, carefully add 3 ml of GENMED cleaning solution (Reagent A), and mix well. Transfer to a pre-chilled 10ml tube. Centrifuge at 4°C for 5 minutes at 300g. Carefully aspirate the supernatant.
[0057] 1.2 Add 500 microliters of GENMED Lysis Solution (Reagent B) and mix well. Transfer to a pre-chilled 1.5 ml centrifuge tube. Vortex vigorously for 15 seconds. Incubate on ice for 30 minutes. Centrifuge at 4°C for 5 minutes at a speed of 16,000 g.
[0058] 1.3 Carefully pipette 500 µl of the supernatant into a new pre-cooled 1.5 ml centrifuge tube. Pipette 10 μl for protein quantification. Immediately store at -70°C or place in an ice bath to continue subsequent operations.
[0059] 2. Assay Preparation
[0060] 2.1 Place the platelet sample in an ice bath.
[0061] 2.2 Set the conditi...
Embodiment 3
[0079] Experimental study on the role of platelet GSK-3β activity in cognitive impairment in patients with type 2 diabetes mellitus (T2DM)
[0080] Experimental methods: (1) Inclusion criteria: Diabetic patients aged >50 years old; no visual and hearing impairments that significantly affect cognitive tests; no history of severe hypoglycemia, diabetic ketosis coma, hyperosmolar coma; no alcohol dependence and psychoactivity History of substance abuse; no brain trauma, no cardiovascular and cerebrovascular events, including cerebral infarction, cerebral hemorrhage, TIA, myocardial infarction, etc.; no other serious physical diseases that affect cognitive tests, such as epilepsy, hypothyroidism, hypoxemia symptoms, etc.; no history of mental illness; voluntary participation in this study, able to cooperate with the examination, and good compliance.
[0081] (2) Study groups: type 2 diabetes without cognitive impairment T2DM group (T2DM), type 2 diabetes with mild cognitive impair...
PUM
Abstract
Description
Claims
Application Information
- R&D Engineer
- R&D Manager
- IP Professional
- Industry Leading Data Capabilities
- Powerful AI technology
- Patent DNA Extraction
Browse by: Latest US Patents, China's latest patents, Technical Efficacy Thesaurus, Application Domain, Technology Topic, Popular Technical Reports.
© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap|About US| Contact US: help@patsnap.com