Preparation and application of human apoptosis factor quantitative antibody chip
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An apoptotic factor and antibody chip technology, applied in the biological field, can solve the problems of small coverage, low throughput, high background, etc., and achieve the effect of preventing errors, high sensitivity, and high sensitivity
Inactive Publication Date: 2013-07-10
GUANGZHOU CINI BIOLOGICAL SCI & TECH
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Problems solved by technology
The existing antibody chip fails to solve the key links such as spotting conditions, carrier treatment and modification, antibody immobilization, and sample processing in the production process of the antibody chip. Therefore, the throughput of product detection is small, the product is unstable, and the specificity is not high. And the reliability is not strong, and because of the small amount of coverage, it is easy to produce false positives or false negatives
The protein extraction solution provided in the antibody chip kit cannot maintain the activity of the protein well, and the extraction rate is low
The detection methods and reagents provided in the antibody kit also have problems such as poor specificity and high background of the experimental results
Method used
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Embodiment 1
[0040] Embodiment 1: the establishment of the quantitative antibody chip preparation method of human TNFα signal transduction pathway apoptotic factor
[0041] 1.1 The basic procedure of antibody chip preparation: design of antibody chip, treatment of glass slide, chemical modification of glass slide, quality inspection of antibody, determination of optimal concentration of capture antibody and detection antibody, microarray spotting antibody chip, antibody The sealing of the chip and the installation of multiple sample fences, the quality control of the antibody chip, packaging and storage?
[0042] 1.2 Antibody chip design: In order to avoid the variation between the points in the chip and improve the accuracy of detection, the Latin square experimental design was applied to the experimental design of chip spotting, which greatly eliminated the inter-row variation and Inter-column variation, reducing the differential reaction between antibody-binding samples, thereby g...
Embodiment 2
[0051] Embodiment 2: Construction human TNFα signal transduction pathway apoptosis factor quantitative antibody chip kit
[0052] 2.1 The components of the kit are as follows:
[0053] 2.1.1 Components of the human TNFα signal transduction pathway apoptosis factor quantitative antibody chip:
[0062] (9) Negative control PBS (concentration 0.01mol / l)?
[0063] (10) Positive control biotin...
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Abstract
The invention relates to a human apoptosis factor quantitative antibody chip and a kit. A buffer liquid capable of being mixed with various standard proteins is prepared; a protein extraction reagent capable of effectively extracting proteins and ensuring that the proteins keep excellent activity is prepared; and whether a color indicator is added to the antibody so as to prevent errors of missing addition or less addition in detection is detected. The method for preparing the human apoptosis factor quantitative antibody chip specifically comprises design of the antibody chip, processing of a glass slide, chemical modification of the glass slide, quality examination of the antibody, determination of optimal concentration of capture antibodies and detection antibodies, micro-array spotting of the antibody chip, sealing of the antibody chip, mounting of a multi-sample fence, quality control of the antibody chip, and packaging and putting in storage. The application of the human apoptosis factor quantitative antibody chip kit is oriented to detect the expression condition of the human apoptosis factor in diseases. The human apoptosis factor quantitative antibody chip provided by the invention is higher in detection flux, better in flexibility, smaller in detection background, higher in specificity, more scientific in operation and more convenient for detection.
Description
technical field [0001] The invention belongs to the field of biotechnology, in particular to the preparation and application of a human cell apoptosis factor quantitative antibody chip. Background technique [0002] Apoptosis is a genetically programmed cellular event that plays an important role in normal tissue development and homeostasis. Defects in the apoptotic program are thought to lead to a variety of human diseases ranging from neurodegenerative and autoimmune diseases to tumors. disease. As a genetic process, apoptosis, like any other developmental process, can be disrupted by mutation, and alterations in the apoptotic pathway are believed to play a critical role in many disease processes, including cancer. For example, the disorder of apoptosis can lead to autoimmune diseases, viral infections, AIDS, cardiovascular diseases, osteoporosis, tumors, neurodegenerative diseases (Parkinson's disease), etc. Research into cancer initiation and progression has tradition...
Claims
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