Test - lowercase jpg
Kaixin micro test
Test probe P100-M3
EL-C1600N100013-B
ELISA test kit principle:

IL-2 is mainly produced by activated T cells and is also required for T cell proliferation, so it is called T Cell Growth Factor (TCGF). The level of IL-2 production reflects the function of T cells, 牪; it is only an important subject of immune regulation, and is closely related to many clinical diseases. CTLL is derived from C57BL/6.犘! The murine killer T lymphocyte cell line, established by Gills in 1977, can only grow in the medium in which IL-2 is present. Therefore, it can be used as an indicator cell to determine the level of IL-2 in the sample to be tested. In addition to CTLL, mitogen-activated T lymphoblasts can also serve as indicator cells for detecting IL-2 activity.

(ii) Method

3H-TdR incorporation method and mtt method can be selected according to laboratory conditions

1. 3H-TdR incorporation method

IL-2 dependent CTLL washed with 10% FCS RPMI1640

2 times, 1000 rpm, 5 min each time, remove IL-2 from the original culture solution

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Adjust the number of living cells 1 × 105 / ml

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Add 100 μl per well to a 96-well flat-bottomed plate

CTLL suspension (1×104/well)

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Add different dilution standard IL-2 and sample to be tested

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37 ° C CO  incubator, culture 18 ~ 24h

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Add 3H-TdR 0.5μci/50μl per well

↓ 4-6h

Harvested on fiberglass paper with a cell harvester

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After drying, transfer to a liquid flash bottle and add 1 ml of scintillation fluid.

Measuring the cpm value of beta rays in a liquid scintillation meter

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Calculation (example)

The cpm values ​​obtained from different concentrations of standard IL-2 and different dilutions of the sample to be tested are plotted. The axis is probit and the abscissa is the log2 dilution factor. From the highest cpm value of 50%, draw a horizontal line, the intersection of the 牦k standard and the oblique line to be examined, and calculate the activity unit of the sample IL-2 to be tested.

If the standard product has a maximum cpm value of 1%/ml, the sample to be tested is 1u/ml at 1:4, and the stock solution is 4μ/ml.

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