Western blot analysis of extracts from Raw 264.7 cells, untreated (-) or treated (+) with LPS (100 ng/mL, 6 hr) and Brefeldin A #9972 (300 ng/mL, last 3 hr of stimulation), using TNF-α (D2D4) XP ® Rabbit mAb (Rodent Specific) (upper) or β-Actin (D6A8) Rabbit mAb #8457 (lower).
Western blot analysis of 1 ng recombinant mouse TNF-α #5178 and 1 ng recombinant rat TNF-α using TNF-α (D2D4) XP ® Rabbit mAb (Rodent Specific).
Immunoprecipitation of TNF-α from Raw 264.7 cell extracts, treated with LPS (100 ng/mL, 6 hr) and Brefeldin A #9972 (300 ng/mL, last 3 hr of stimulation), using Rabbit (DA1E) mAb IgG XP ® Isotype Control #3900 (lane 2) or TNF-α (D2D4) XP ® Rabbit mAb (Rodent Specific) (lane 3). Lane 1 is 10% input. Western blot analysis was performed using TNF-α (D2D4) XP ® Rabbit mAb (Rodent Specific). Mouse Anti-rabbit IgG (Conformation Specific) (L27A9) #3678 and Anti-mouse IgG, HRP-linked Antibody #7076 were used as sencondary antibodies.
Confocal immunofluorescent analysis of Raw 264.7 cells, treated with LPS (100 ng/mL, 6 hr; left) or untreated (right), using TNF-α (D2D4) XP ® Rabbit mAb (Rodent Specific) (green). Actin filaments were labeled with DY-554 phalloidin (red). Blue pseudocolor = DRAQ5 ® #4084 (fluorescent DNA dye).
Flow cytometric analysis of Raw 264.7 cells, untreated (blue) or treated with LPS (100 ng/ml, 6 hr) and Brefeldin A #9972 (300 ng/ml, last 3 hr of treatment) (green), using TNF-α (D2D4) XP ® Rabbit mAb (Rodent Specific). Anti-rabbit IgG (H+L), F(ab') 2 Fragment (Alexa Fluor ® 647 Conjugate) #4414 was used as a secondary antibody.