Anti diap1 antibody

After activation with LPS, isolated primary human neutrophils and HL-60-derived neutrophils (control and TLR10 knockdown) were lysed in RIPA buffer (Sigma, St. Louis, MO, USA) with protease inhibitor cocktail. Immunoblots were prepared by anti-TLR10 antibody (1:600, 1 mg/mL; Abcam, Cambridge, MA, USA), anti-Diap1 antibody (1:500, Cell Signaling Tech, Danvers, MA, USA), anti-ARP3 antibody (1:400, Cell Signaling Tech, Danvers, MA, USA). Anti-β-actin (1:1,000, Abcam, Cambridge, MA, USA) was used as loading control. HRP-conjugated anti-goat or anti-rabbit (1:2000; DAKO, Burlington, VT, Canada) was used as the secondary antibody. Hybridization signals were detected with Amersham ECL Western blotting detection reagents (GE Healthcare, Mississauga, ON, Canada). Blots were digitally imaged and contrast adjustments were applied to all parts of the figure in an unbiased manner. Represented lanes indicate only parts of the blot for better visualization. Quantification of bands from immuno­blots was performed with densitometry in Adobe Photoshop CS6 (San Jose, CA, USA).