Mouse pdl1 elisa kit

The intracranial injection of GL261 tumor cells was performed as previously described [29 (link)] in compliance with UAB animal care policy. The procedure of CFS collection was reported [31 ] and performed according to UAB animal care policy. The PDL1 level in CSF samples was analyzed by using the Mouse PDL1 ELISA Kit (Boster, Pleasanton, CA, USA). Mice were euthanized by using an IP injection of ketamine (500 mg/150 uL) in combination with an alpha 2-adrenergic receptor agonist followed by bilateral thoracotomy. The euthanasia method is approved by UAB Animal Care policy and by the AVMA Panel on Euthanasia, 2013 Edition.

Cells were plated at 2×10⁶ per P10 plate. Fresh media (6 ml) was added to the cells and collected after 24 hours. Collected media was spun at 4°C, 5 min, 1200 rpm to remove any cells and cell debris. After debris removal, the protein content in the media was concentrated 6 to 8-fold by using the 10K protein concentrator PES (Thermo Scientific) at 4°C. The protein concentrate was reconstituted in Cell Lysis Buffer (Cell Signaling Technology) and analyzed by Western blot.
Antibodies, Reagents, and Drugs. HIF1A, Lamin A/C, hPDL1, anti-biotin HRP-linked, and anti-rabbit IgG1 HRP-linked antibodies were purchased from Cell Signaling Technologies (Danvers, MA, USA); Actin, HuR 3A2, PDL1, donkey anti-goat IgG-HRP, goat anti-mouse IgG1-HRP antibodies were from Santa Cruz Biotechnology (Dallas, Texas, USA); the mPDL1/B7-H1 antibody was from B&D systems (Minneapolis, MN, USA); the alpha Tubulin antibody was from Sigma (Sigma-Aldrich). Mouse PDL1 ELISA Kit was purchased from Boster (Pleasanton, CA, USA). MLN4924 was purchased from Active Biochem (Kowloon Bay, Hong Kong); the PD1/PDL1-inhibitor-1 was purchased from Cayman Chemical (Ann Arbor, Michigan, USA).