CD90/Thy1 Antibody (G7) - Azide and BSA Free

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• CD90/Thy1 Antibody (G7)

  Name: Kevin Thorburn

  Application: Immunocytochemistry

  Sample Tested: Cortical neurons

  Species: Mouse

  Verified Customer | Posted 07/23/2019

  Immunopanned primary cortical neurons isolated from postnatal day 7 CD1 mice.

  Cortices from postnatal day 7 CD1 mice were dissected and dissociated according to the methods of LC Foo - Cold Spring Harbor 2013. The cell suspension was sequentially incubated on petri dishes coated with antibodies targeting CD45, O4, Thy1/CD90 and Itgb5. Cells adhered to the CD90 dish were removed with trypsin and replated in a neurobasal-based media containing 2% B-27 plus. Cells were fixed with PFA for 10 min 36h after being plated. The cells were then washed 3 times with D-PBS -Sigma D8662- and blocked for 30 minutes in D-PBS containing 0.2 percent Triton X100 and 10 percent donkey serum. The cells were then incubated overnight at 4 degrees with the primary antibody at 1 in 500 dilution in D-PBS containing 0.2 percent triton, 2 percent donkey serum and 2 percent bovine serum albumin - heat shock fraction. The following day the cells were washed twice with D-PBS and then incubated with Alexa Fluor secondaries - 1 in 500 dilution in the same antibody buffer described above - from ThermoFisher for 30min at room temperature. Cells were then washed 3 times with D-PBS and imaged.

  

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