Recombinant human nerve growth factor - β (β - NGF)

Product attributes:

Background:

Nerve growth factor（NGF）It is crucial for the development and maintenance of the sympathetic and sensory nervous systems.NGFIt has been confirmed to be a complex composed of three peptides called alpha, beta, and gamma subunits. Among them, the β subunit, namely β-NGFIt is a kind of26.9 kDaThe protein contains 241 A residue that participates in the survival and differentiation of neurons. In addition, β-NGFstillTRKAThe ligand of the receptor, andTRKAReceptors are involved in the differentiation and development of pain and temperature sensing neurons.

Tag:

1. Mix the cell culture medium with EdU solution in a ratio of 500:1 to prepare 2 × EdU labeled culture medium.

2. Preheat the 2 × EdU labeled culture medium in advance, and then mix 150 μ L of 2 × EdU labeled culture medium with an equal volume of cell culture medium to obtain a 1 × EdU solution.

[Note]: ① It is not recommended to replace all the culture media, as this may affect the speed of cell proliferation. ② It is recommended to use the prepared culture medium immediately, and the dosage should not exceed the cells.

3. Incubate the cells for 2 hours and discard the culture medium.

【 Note 】: ① The cultivation time depends on the proliferation rate and growth status of the cells. ② Most tumor cells and adherent cell lines can be incubated for 2 hours.

4. Wash the cells twice with 1xPBS for 5 minutes each time to remove EdU residues that have not been incorporated into DNA.

[Note]: For cells that are not firmly attached to the wall, the cleaning intensity can be reduced.

Experimental procedure:

1. Inoculate cells at a density of 104-105 cells/well into a 96 well plate and add 100 μ L of culture medium, with or without the test compound. Cultivate cells in a CO2 incubator at 37 ℃ for 24 hours.

2. Add 10 μ L of different concentrations of the test substance to the plate.

3. Cultivate the culture plate in the incubator for an appropriate period of time (such as 6, 12, 24, or 48 hours).

4. Add 10 μ L of CCK-8 solution to each well of the plate. Be careful not to introduce bubbles into the hole.

5. Incubate the plate in the incubator for 1-4 hours.

6. Gently mix on the fixed orbit oscillator before reading the board.

7. Measure the absorbance at 450nm using an enzyme-linked immunosorbent assay (ELISA) reader.

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