Cell Migration Assay Service

Instruction manual

First Edition (Revised on April, 2016)

【Service content】

The purpose of this experiment is to detect cell migration ability. Add the cell to be studied into the upper chamber of transwell, and add FBS with a high concentration into the lower. The cells will penetrate polycarbonate membranes under the influence of FBS. Record the number of transmembrane cells. The number of the cells in different treatment groups are different, which shows the different migration ability.

【Protocol& Period】

Period: 1~2 weeks

Protocol:

(1) Prepare single cell suspension: Digest cells after transfection with 0.25% of trypsin, then collects it and centrifuge it at 1000rpm for 5 minutes. Abandon the supernatant and resuspend cells with serum free DMEM medium. Count and adjust density to 1x10e6/ ml.

(2) Inoculate cells: Add 200ul monocyte suspension into the upper chamber of transwell, and add 600ul DMEM culture including 30% FBS into the lower chamber. Each group was set up with three wells. Put it into rsbiotech with the condition of 5％CO2 and 37 degrees for 72 hours.

(3) Fix and dye: Take out the Transwell and absorb the medium of the upper chamber. Wash it three times with PBS buffer. Use sterile wet cotton swab to gently wipe off the uninvaded cells and Martrigel glue. Fix it with 95% alcohol for 30 minutes and dye it with Crystal violet for 10minutes. Wash it more than 3 times and then dry in the air.

(4) Take photos and count: Examined it under a 20 times optical microscope. Count and calculate the average amount. And the experimental data were repeated three times.

【Provided by customer】

1. Cell lines for assay or disposing (Our company can provide common used cell lines).

2. Drugs and experiment scheme for cell experiment.

【Service results】

1. Providing the experimental data and images.

2. Providing the test report.