Novus Biologicals
Anti-LC3
NB100-2331
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Our research focuses on autophagy and hence we do a lot of analyses related to LC3, an autophagy marker. We have tested this antibody with a wide range of cell types including Sy5Y, HEK293, NIH3T3, MEFs, N2a neuroblastoma, Q7/7 striatal cells and many more. PFA fixation works well and methanol works okay as well. This antibody gives a very specific signal and was optimized and confirmed with lysosomal inhibitors (where LC3 levels are reported to increase a lot). We have tried a couple of other LC3 antibodies from different companies; although many antibodies are good for Western blot, immunofluorescence, and immunoprecipitation, by far this was the best among the very few ones which were good for immunofluorescence.
Immunofluorescence
Sy5Y, HEK293, NIH3T3, N2a, Q7/7, MEFs
1:100, 1 hour, room temperature in 0.1% BSA
0.2% milk, 2% serum, 0.1M glycine, 1%BSA, 0.01%Tx-100 in PBS
1:300, 1 hour, room temperature rabbit conjugated texas-red, Cy5 and Alexa488 can be used.
NA
Fluorescence
LC3 appears in punctate pattern in PFA fixed coverslips.
None
Short incubation, specificity
Rabbit polyclonal, cannot be co-stained with ease with other rabbit polyclonal antibodies.
Great antibody for immunofluorescence, works in wide types of samples.