Confocal immunofluorescent analysis of HeLa cells, untreated (left) or treated with Bafilomycin A1 (100 nM, 18 hr; right), using SQSTM1/p62 (D10E10) Rabbit mAb (IF Preferred) #7695 (green). Actin filaments were labeled with DyLight™ 554 Phalloidin #13054. Blue pseudocolor = DRAQ5® #4084 (fluorescent DNA dye).
Confocal immunofluorescent analysis of HeLa cells, untreated (left) or treated with Bafilomycin A1 (200 nM, 18 hr; right), using NBR1 (D2E6) Rabbit mAb #9891 (green) and β-Actin (8H10D10) Mouse mAb #3700 (red). Blue pseudocolor = DRAQ5® #4084 (fluorescent DNA dye).
Chemical structure of Bafilomycin A1.
Bafilomycin A1 is supplied as a lyophilized powder. For a 1 mM stock, reconstitute the 100 μg in 160.56 μl DMSO. Working concentrations and length of treatment can vary depending on the desired effect, but it is typically used at 10 nM-1 μM for up to 18 hr.
Store lyophilized or in solution at -20ºC, desiccated. Protect from light. In lyophilized form, the chemical is stable for 24 months. Once in solution, use within 3 months to prevent loss of potency. Aliquot to avoid multiple freeze/thaw cycles.
Isolated from Streptomyces griseus, bafilomycin A1 is a macrolide antibiotic that reversibly inhibits late phase autophagy (1,2). By inhibiting H+-ATPase, bafilomycin A1 prevents re-acidification of synaptic vesicles following exocytosis and thereby blocks fusion of autophagosomes and lysosomes (2-5). Research studies demonstrate that bafilomycin A1 can also inhibit cell proliferation (4).
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DRAQ5 is a registered trademark of Biostatus Limited.
DyLight is a trademark of Thermo Fisher Scientific, Inc. and its subsidiaries.