Looking for breakthrough ideas for innovation challenges? Try Patsnap Eureka!

NF-kappaB RelA/p65 protein loci specificity phosphorylation diagnosis kit

A technology of p65 and kits, applied in fluorescence/phosphorescence, biological testing, instruments, etc., can solve problems such as less success

Active Publication Date: 2019-10-11
湖南莱拓福生物科技有限公司
View PDF4 Cites 0 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

So far, nearly a thousand NF-κB inhibitors have been developed. Although some inhibitors have achieved promising results in experimental treatments, they are rarely successful in clinical practice.

Method used

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
View more

Image

Smart Image Click on the blue labels to locate them in the text.
Viewing Examples
Smart Image
  • NF-kappaB RelA/p65 protein loci specificity phosphorylation diagnosis kit
  • NF-kappaB RelA/p65 protein loci specificity phosphorylation diagnosis kit
  • NF-kappaB RelA/p65 protein loci specificity phosphorylation diagnosis kit

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0036] Example 1: Preparation of Sandwich Time-Resolved Fluorescence Immunoassay (TRFIA) Kit for Detection of NF-κB RelA / p65 Ser276 and Ser536 Phosphorylated Proteins

[0037] In this example, the sandwich time-resolved fluorescent immunoassay (TRFIA) kit includes:

[0038] Plates or beads coated with specific capture monoclonal antibodies;

[0039]Biotin or other labeled specific detection antibody from another species;

[0040] Coating buffer (1×PBS, pH7.3±0.1);

[0041] Sample diluent (1×PBS, 1%BSA, 0.05%NaN3, pH 7.3±0.1);

[0042] Blocking solution (1×PBS, 3% BSA);

[0043] Eluent PBST (1×PBS, 0.05% Tween-20);

[0044] Streptavidin-Eu / HRP / ALP;

[0045] The biotin labeling kit was purchased from Pierce Company. Streptavidin-Eu was purchased from Perkin Elmer. The time-resolved fluorescent immunoassay (TRFIA) plate for detection was prepared by ourselves, and the capture antibody was diluted to 7-10 μg / ml with coating buffer, and 100 μl of the above-mentioned Antibod...

Embodiment 2

[0049] Example 2: Detection of phosphorylation levels of NF-κB RelA / p65 Ser276 and Ser536 in tumor tissues of lung cancer patients using a sandwich time-resolved fluorescent immunoassay (TRFIA) kit

[0050] Detection Time-resolved fluorescence immunoassay plate preparation: Dilute the capture antibody to 7-10 μg / ml with coating buffer, add 100 μl of the above antibody dilution to each well of the time-resolved fluorescence immunoassay (TRFIA) plate, and leave overnight at 4ºC Take it out and place it at 37ºC for 30 minutes; shake off the liquid and wash 5 times with PBST; air dry, add 100 μl blocking solution to each well, and keep at 37ºC for 2 hours; shake off the liquid and wash twice with PBS; air dry at room temperature 2 hours; vacuum seal with foil, store at 4ºC, and set aside.

[0051] Detection of tissue samples: Tumor tissues were prepared with standard methods for soluble protein solution, diluted 1:10 with PBS, and 100 μl of sample diluent or standard was added to ...

Embodiment 3

[0055] Example 3: Detection of Phosphorylation Levels of NF-κB RelA / p65 Ser276 and Ser536 in Tumor Tissues of Colon Cancer Patients Using Sandwich Time-Resolved Fluorescence Immunoassay (TRFIA) Kit

[0056] Detection Time-resolved fluorescence immunoassay plate preparation: Dilute the capture antibody to 7-10 μg / ml with coating buffer, add 100 μl of the above antibody dilution to each well of the time-resolved fluorescence immunoassay (TRFIA) plate, and leave overnight at 4ºC Take it out and place it at 37ºC for 30 minutes; shake off the liquid and wash 5 times with PBST; air dry, add 100 μl blocking solution to each well, and keep at 37ºC for 2 hours; shake off the liquid and wash twice with PBS; air dry at room temperature 2 hours; vacuum seal with foil, store at 4ºC, and set aside.

[0057] Detection of tissue samples: Tumor tissues were prepared with standard methods for soluble protein solution, diluted 1:10 with PBS, and 100 μl of sample diluent or standard was added to ...

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

PUM

No PUM Login to View More

Abstract

The invention relates to the technical field of the medical biology, and specifically relates to a kit for NF-kappaB RelA / p65 protein loci specificity phosphorylation detection. Specifically, the kitcan specifically detect the phosphorylation state of the NF-kappaB RelA / p65 protein Ser276 and Ser536 site so as to guide the NF-kappaB inhibitor targeting anti-tumor drug therapy or RelA / p65 S276A / S536D; and the kit has important clinical application value.

Description

technical field [0001] The invention relates to the field of medical biotechnology, in particular to a kit for specifically detecting phosphorylation of human NF-κB RelA / p65 protein site, which is used for detecting Ser276 of NF-κB RelA / p65 protein in various human tumor tissues The phosphorylation status of Ser536 site is used for targeted anti-tumor drug therapy and anti-tumor gene therapy. The kit involves immunoturbidimetry, enzyme-linked immunoassay, time-resolved immunofluorescence analysis (TRFIA) method, chemiluminescence method and electrochemiluminescence method based on sandwich ELISA. The kit also involves immunohistochemistry (IHC) and immunocytochemistry (ICC) methods. Background technique [0002] The eukaryotic transcription factor protein family NF-κB RelA / p65 (NF-κB RelA / p65 gene in GenBank accession number L19067.1) is a key regulatory protein in immune response and inflammatory response. The mammalian NF-κB / Rel family includes five subunits, which are R...

Claims

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

Application Information

Patent Timeline
no application Login to View More
IPC IPC(8): G01N33/577G01N33/68G01N21/64
CPCG01N33/577G01N33/6893G01N21/6408G01N21/6486
Inventor 曹哲
Owner 湖南莱拓福生物科技有限公司
Who we serve
  • R&D Engineer
  • R&D Manager
  • IP Professional
Why Patsnap Eureka
  • Industry Leading Data Capabilities
  • Powerful AI technology
  • Patent DNA Extraction
Social media
Patsnap Eureka Blog
Learn More
PatSnap group products

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