Anti p eef2k s366

Antigen retrieval was conducted by preheating in sodium citrate buffer (pH 6.0) for 30 minutes at 95°C in a pressure cooker. Tissue was blocked in 1% BSA solution for 2 hours or in M.O.M blocking solution (Vector Laboratories, MKB 2213) for 1 hour at room temperature, followed by blocking in 1% BSA solution for 2 hours. Primary antibodies were applied overnight at 4°C in a humidified chamber, and secondary antibodies (Alexa-Fluor 594 : Invitrogen, A11032; Alexa-Fluor 488 : Invitrogen, A11034; Alexa-Fluor 594 : Invitrogen, A11037; Alexa 647: Invitrogen, A-21244) were applied for 1 hour at room temperature. All IF slides were mounted using ProLong gold mounting media with DAPI (Fisher Scientific, P36935). IF slides were scanned using an Aperio Scanscope FL (Leica Biosystems). The following primary antibodies were used for IF study: anti-Cytokeratin 5 (Biolegend Inc, PRB-160P), anti-puromycin (EMD Millipore, MABE343), anti-p-eEF2K(S366) (Cell Signaling Technology, 3691), anti-ARID1A (Sigma, HPA005456), anti-Cytokeratin 5 (Abcam, ab52635). Image analysis was done using semiautomated image analysis software (HALO, Indica Labs).

Antigen retrieval was conducted by preheating in sodium citrate buffer (pH 6.0) for 30 minutes at 95°C in a pressure cooker. Tissue was blocked in 1% BSA solution for 2 hours or in M.O.M blocking solution (Vector Laboratories, MKB 2213) for 1 hour at room temperature, followed by blocking in 1% BSA solution for 2 hours. Primary antibodies were applied overnight at 4°C in a humidified chamber, and secondary antibodies (Alexa-Fluor 594 : Invitrogen, A11032; Alexa-Fluor 488 : Invitrogen, A11034; Alexa-Fluor 594 : Invitrogen, A11037; Alexa 647: Invitrogen, A-21244) were applied for 1 hour at room temperature. All IF slides were mounted using ProLong gold mounting media with DAPI (Fisher Scientific, P36935). IF slides were scanned using an Aperio Scanscope FL (Leica Biosystems). The following primary antibodies were used for IF study: anti-Cytokeratin 5 (Biolegend Inc, PRB-160P), anti-puromycin (EMD Millipore, MABE343), anti-p-eEF2K(S366) (Cell Signaling Technology, 3691), anti-ARID1A (Sigma, HPA005456), anti-Cytokeratin 5 (Abcam, ab52635). Image analysis was done using semiautomated image analysis software (HALO, Indica Labs).