Anti mouse igg hrp 7076s

Equal concentration protein from cell lysates and CM were separated by 12% SDS/PAGE gels. The proteins were transferred to nitrocellulose or PVDF membranes and blocked with 5% skim milk. The membranes were probed using anti‐human ALDH1A3 (OTI4E8; OriGene Technologies, Rockville, MD, USA), tPA (ab227069; Abcam, Toronto, Canada), uPA (ab24121; Abcam), and PAI‐2 (ab47742; Abcam) antibodies. Anti‐rabbit IgG‐HRP (7074S; Cell Signaling Technology) secondary antibody was used for tPA, uPA, and PAI‐2 while anti‐mouse IgG‐HRP (7076S; Cell Signaling Technology, Danvers, MA, USA) secondary antibody was used for ALDH1A3. Immuno‐reactive proteins were detected by chemiluminescence (using Clarity ECL blotting substrate (Bio‐Rad)) and visualized with images captured with a ChemiDoc Imager (Bio‐Rad). Total protein was used as a loading control.

Anti-CX3CL1 antibody (ab25088), anti-CX3CR1 antibody (ab8021), anti-CX3CL1 antibody (ab25088), anti-CX3CR1 antibody (ab8020), anti-CD45 antibody (ab8216), anti-MHC class II antibody (ab23990), anti-Runx1 antibody (ab23980), anti-Iba1 antibody (ab5076), anti-NSE antibody (ab53025), and anti-NeuN (ab104224) were from Abcam (USA). Anti-MCPIP antibody (sc-515275) and β-tubulin (sc-9014) were obtained from Santa Cruz Biotechnology (USA). Anti-C/EBPα antibody (2295), anti-rabbit-IgG-HRP (7074s), and anti-mouse-IgG-HRP (7076s) were obtained from the Cell Signaling Technology (USA). Alexa Fluor-488 donkey anti-rabbit IgG antibody (A21206), Alexa Fluor-555 donkey anti-mouse IgG antibody (A31570), Alexa Fluor-555 donkey anti-goat IgG antibody (A21432), and Alexa Fluor-633 donkey anti-goat IgG antibody (A21082) were from the Life Technologies. RBFOX3/NeuN antibody [Alexa Fluor-405] (NBP1-77686AF405) was from Novus (USA).

Antibodies against human lactadherin (monoclonal sc-8029), CD63 (sc-5275 or ab68418), TSG101 (sc-7964), calnexin (sc-46669), GAPDH (sc-47724) and β-actin (sc-47778) were purchased from Santa Cruz Biotechnology, Inc. or Abcam. A policlonal antibody against lactadherin was also used (AF2767-SP) and purchased from RyD Systems. Antibody against human flotillin 1 (18634S) was purchased from Cell Signaling Technology (Danvers, MA). Alexa Fluor 488-conjugated anti-lactadherin antibody (sc-8029) was also purchased from Santa Cruz Biotechnology Inc. Anti-rabbit IgG-HRP (7074S) and anti-mouse IgG-HRP (7076S) secondary antibodies were purchased from Cell Signaling Technology. Goat anti-rabbit IgG-800CW (#92,532,211) and goat anti-mouse IgG-680LT (#925–68,020) secondary antibodies were obtained from Licor.