Application of Prohibitin protein antibody to preparing kit for diagnosing senile dementia

A kit, age-related technology, applied in biological tests, material inspection products, etc., can solve the problems of synapse loss, cholesterol transport dysfunction, time-consuming and labor-intensive, etc.

Inactive Publication Date: 2012-10-03
CHANGSHU CITY YUSHAN GREEN TEA
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

③ApoE gene: ApoE causes cholesterol transport dysfunction in the process of lipid transport, and finally leads to synaptic loss
As mentioned earlier, sequences can be determined from MS data, but this is a time-consuming and labor-intensive task when analyzing hundreds or thousands of spectra from scratch

Method used

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  • Application of Prohibitin protein antibody to preparing kit for diagnosing senile dementia
  • Application of Prohibitin protein antibody to preparing kit for diagnosing senile dementia

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0036] Embodiment 1, the preparation of AD cell model sample:

[0037] 1. Cell culture:

[0038]Take out the frozen SH-SYSY cells from liquid nitrogen for resuscitation, quickly put them into a 37°C water bath, dissolve for about 1min, and dilute the 15% fetal bovine serum culture medium returned to room temperature to 5 times the original volume (8-10ml), and centrifuge at 1000rpm at low speed , 10min, discard the supernatant, use 15ml of 15% fetal bovine serum medium for precipitation, 37℃, 5% CO 2 to cultivate. Invert the microscope to observe the growth of the cells. When the cells cover the bottom of the culture flask, replace the medium (about 48 hours), pour out the old culture medium, and use 0.25% trypsin to act on the cells for 1 min. Stop digestion immediately when the cells become round. Then use the direct pipetting method of culture medium to subculture (direct pipetting is also possible). Properly freeze and preserve the cells in good growth state, the meth...

Embodiment 2

[0046] Example 2, Screening of Differential Proteins

[0047] 1. First dimension isoelectric focusing electrophoresis and equilibration

[0048] Mix the sample with a hydration solution (7mol / L urea, 2mol / L thiourea, 4% CHAPS, 65mmol / L DTT, 0.2% IPGBuffer, trace bromophenol blue) in a total volume of 500 μL, with a total protein content of 200 μg , into the focusing pan, and place the IPG dry strip glue side down in the focusing pan, and then conduct isoelectric focusing in the ProteanIEF Cell isoelectric focusing instrument. The parameters were set as follows: passive hydration at 17°C for 16 h, focusing at 100 V for 5 h, focusing at 250 V for 3 h, focusing at 500 V for 2 h, focusing at 1 000 V for 2 h, focusing at 10 000 V for 5 h, and the total voltage-time product was 60 Focusing was terminated after 000 Vh. After electrophoresis, the gel strips were taken out for washing and equilibrated in DTT and iodoacetamide for 15 min each time.

[0049] 2. Second dimension SDS-...

Embodiment 3

[0060] Example 3, Western blot verification of differential expression of Prohibitin protein

[0061] In order to confirm the differential expression of Prohibitin protein, 3 cases of serum samples from patients diagnosed with AD were taken, and 1 case of normal serum samples confirmed to be suffering from AD was taken as a control. After routine sample processing, Western blot detection was performed. The experimental steps are roughly as follows: load 40 mg of protein on electrophoresis (SDS-PAGE gel), and transfer to Immobilon-P membrane (Millipore Corporation) at a voltage of 150V. After blocking with 5% skimmed milk powder at room temperature for 1.5 h, the primary antibody was added. The primary antibodies were anti-β-Actin antibody and anti-Prohibitin antibody (1:1000; Abcam Company), and the next day, the secondary anti-horseradish peroxidase antibody was added and stained by ECL chemiluminescence (ECL kit, Amersham Company) . The Western blot verification conclus...

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Abstract

The invention discloses an oligopeptide with breast cancer resisting activity. A sequence of the oligopeptide is His-Asn-Tyr-Gly-Phe-Val-Pro-Ser-Leu. The oligopeptide with low concentration also can selectively suppress growth of extracorporeal breast cancer cells and does not affect growth of normal mammary gland cells. Extracorporeal tests also show that the oligopeptide has a high suppression effect for the growth and proliferation of cancer cells, and obvious dose and effect relation is displayed. Owing to the safe, efficient and ideal anti-cancer effect, the oligopeptide can be used for preparing anti-cancer medicines and has a good application prospect.

Description

technical field [0001] The invention relates to the application of Prohibitin protein antibody in the preparation of a kit for diagnosing senile dementia. Background technique [0002] At present, the aging rate of the world's population is accelerating day by day, and the average life expectancy of the population in economically developed countries has reached or is close to 80 years old. The average life expectancy of the population in my country is 70 years old, and the number of people over 60 years old has reached 130 million. It is estimated that in this century, aging and aging-related diseases such as senile dementia will become important factors affecting human health and quality of life. In western countries and big cities such as Shanghai in my country, senile dementia has become the fourth cause of death after heart disease, tumor and stroke. [0003] Senile dementia, also known as Alzheimer's dementia (Alzheimer's disease, AD), was first reported by German neu...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/68G01N33/53
Inventor 吴健
Owner CHANGSHU CITY YUSHAN GREEN TEA
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