Recombinant human bone morphogenetic protein 3 (BMP-3)

Product attributes:

Background:

BMP-3Also known as osteogenic factor, it is a high content in adult bonesBMPAt least15Related to structure and functionBMPOne of them belongs toTGF-Members of the β superfamily.TGF-Members of the β family are key regulatory factors for cell proliferation, differentiation, matrix synthesis, and apoptosis. As its name suggests,BMPsCan initiate, promote, and regulate the development, growth, and remodeling of bone and cartilage. Apart from thisBMPsAlso involved in the prenatal development, postpartum growth, remodeling, and maintenance of various other tissues and organs.BMP-3Abundant in adult bones, followed by fetal cartilage.BMP-3By activating the signaling cascade and antagonizing the signaling pathway that promotes osteoclastogenes, osteogenesis and bone formation are inhibited.

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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