Rat igg2a apc isotype control

The polyclonal antibodies αN2 and αC6 for PKN1 were prepared as described^{69 (link)}. The polyclonal antibody αNUS for detection of PKN3 was prepared as described^{70 (link)}. Anti PRK2 antibody for PKN2 was purchased from BD Transduction Laboratories, and anti phospho-PRK1 (Thr774)/PRK2 (Thr816) antibody was from Cell Signaling Technology. Annexin V-APC and 7-ADD were purchased from BD Biosciences. The mAbs used for flow cytometric analyses for cell surface antigens were purchased from BD Biosciences, eBioscience or BioLegend. Anti-mouse EDG-1/S1PR1-APC MAb (Clone 713412) and Rat IgG2A-APC Isotype Control were purchased from R&D Systems and used for S1PR1 receptor assay. Anti CD3 rabbit monoclonal antibody (clone SP7; Abcam), an anti B220/CD45R rat monoclonal antibody (clone RA3-6B2; Abcam) and an anti MAC-2/Galectin-3 rat monoclonal antibody (clone M3/38; Cedarlane) were used for immunohistochemical analysis.

SDC4 transfectants, established in K562 cells (ATCC, Manassas, VA, USA; cat. no. CCL-243), were created as described previously [18 (link)]. Stable SDC4 transfectants were selected by measuring SDC4 expression with flow cytometry using APC-labeled SDC4 antibodies (R&D Systems, Minneapolis, MN, USA; SDC4: monoclonal rat IgG2a clone #336304, cat. no. FAB29181A [20 (link),54 (link)]), along with the respective isotype controls (R&D Systems; rat IgG2a APC isotype control, cat. no. IC006A).
L929 (Merck KGaA, Darmstadt, Germany; cat. no. 85011425-1VL) and RAW 264.7 cells (ATCC, Manassas, VA, USA, cat. no. TIB-71) were grown in DMEM, 10% FBS at 37 °C, 5% CO₂.