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ELISA (Enzyme-Linked Immunosorbent Assay) kit for direct detection of activity of human stromal cell-derived factor-1 (SDF-1)

A technology of SDF-1 and stromal cells, which can be used in biological testing, measuring devices, material inspection products, etc., and can solve the problems of high price and affecting the wide application of kit products

Inactive Publication Date: 2011-09-14
ABZYME BIOTECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, the ELISA kits for detecting SDF-1 activity on the market are mainly foreign well-known brands such as R&D, and their prices are extremely expensive, which seriously affects the wide application of kit products in scientific research and production.

Method used

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  • ELISA (Enzyme-Linked Immunosorbent Assay) kit for direct detection of activity of human stromal cell-derived factor-1 (SDF-1)

Examples

Experimental program
Comparison scheme
Effect test

Embodiment

[0023] 1. Coated with SDF-1:

[0024] The target protein SDF-1 was coated on a 96-well microtiter plate at a concentration of 3000nM, and was sequentially diluted in a ratio of 1:3 (PBS was used for the diluent) to form 10 sample gradients of different concentrations, and at the same time, the PBS Instead of samples, as a blank control, 100 μl / well, each concentration was repeated twice. overnight at 4°C.

[0025] Use PBST (PBS+0.5‰Tween 20) to wash 3 times and let stand for 5min each time. Pat dry one last time.

[0026] 2. Closed treatment:

[0027] Freshly prepare 2% gelatin (prepared with PBS), 300 μl / well, and incubate at 37° C. for 2 hr.

[0028] Wash 3 times with PBST, let stand for 5min each time; pat dry for the last time.

[0029] 3. SDF-1 antibody response:

[0030] Anti-SDF-1 goat polyclonal antibody (Santa Cruz Biotechnology, Inc.) was diluted 1:500 with PBS, 100 μl / well, and incubated at 37° C. for 1 hr.

[0031] Wash 3 times with PBST, let stand for 5min ...

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Abstract

The invention relates to an ELISA (Enzyme-Linked Immunosorbent Assay) kit for direct detection of activity of a human stromal cell-derived factor-1 (SDF-1). A method for detecting the activity of the SDF-1 by using the ELISA kit comprises the steps of: coating SDF-1 on an ELISA plate, treating by using a sealing solution in a sealing manner, adding a proper amount of anti-SDF-1 antibodies and a corresponding second antibody, and adding a corresponding developing solution; when the color changes obviously, adding a stopping solution for stopping; measuring an OD (Outside Diameter) value on a microplate reader, and judging the activity of the SDF-1 according to the magnitude of the OD value. The ELISA kit can be used for rapidly and conveniently detecting the activity of the SDF-1 directly, and detecting a plurality of SDF-1 samples in one step in an actual utilization process. The chart in the specification is an SDF-1beta ELISA activity detection reaction chart, the highest concentration of the SDF-1beta is 3,000nM, the lowest concentration of the SDF-1beta is 0.15nM, and EC50 (Encephalitozoon Cuniculi 50) is 80nM.

Description

technical field [0001] The invention relates to an ELISA kit for directly detecting the activity of human stromal cell-derived factor-1 (SDF-1). technical background [0002] SDF-1, also known as CXCL12, is a member of the chemokine protein family. SDF-1 is often used as a specific biological indicator for clinical diagnosis in inflammation, tumors and other malignant lesions. Therefore, it can be detected efficiently and sensitively. The activity of SDF-1 is extremely important in the process of scientific research and clinical diagnosis. [0003] The methods for detecting SDF-1 activity are mainly immunological methods, including enzyme-linked immunosorbent assay (ELISA), immunohistochemical method, western blotting (WB) and so on. Each method has certain advantages and disadvantages, and the corresponding experimental method should be selected according to the purpose of the experiment. The use of direct ELISA to detect SDF-1 activity has the advantages of high efficien...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/68G01N33/543
Inventor 柴笑梅朱月珍杨宣武卢守英江永海
Owner ABZYME BIOTECH
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