Rabbit Monoclonal LC3B Antibody (1251A)

Western Blot: LC3B/MAP1LC3B Antibody (1251A) [NBP2-46892] - Western Blot image of monoclonal anti-LC3B (Clone 1251D). HeLa and Neuro2A cells were treated with or without 50 uM chloroquine for 24 hours as indicated. Whole cell protein was then separated on a 4-15% gel by SDS-PAGE, transferred to 0.2 um PVDF membrane for 30 min and blocked in 5% non-fat milk in TBST. The membrane was probed with 2 ug/ml anti-LC3B in 1% milk, and detected with an anti-rabbit HRP secondary antibody using chemiluminescence. Note the accumulation of LC3 II upon chloroquine treatment.

Immunocytochemistry/Immunofluorescence: LC3B/MAP1LC3B Antibody (1251A) [NBP2-46892] - HeLa cells were treated with Chlorquine for 24 hours prior to fixation, permeabilization and incubation with anti-LC3B (1251A) and anti tubulin (NB100-690) antibodies. Image enlargement shows the accumulation of LC3 (green) on autophagosomes in response to chloroquine treatment. Tubulin staining is shown in red and DNA is counterstained with DAPI (blue).

Immunohistochemistry-Paraffin: LC3B/MAP1LC3B Antibody (1251A) [NBP2-46892] - IHC analysis of a formalin fixed and paraffin embedded tissue section of normal mouse brain using rabbit monoclonal LC3B (1251A) antibody at 1:100 dilution with HRP-DAB detection. The antibody generated a weak diffused cytoplasmic staining in most of the cells but some cells, especially within empty areas on the section, showed punctate signal also which signifies the presence of autophagy in those areas.

Immunocytochemistry/Immunofluorescence: LC3B/MAP1LC3B Antibody (1251A) [NBP2-46892] - HeLa cells were treated with 50 uM Chloroquine for 24 hour prior to fixation in 10% buffered formalin for 10 min. Cells were permeabilized in 0.1% Triton X-100 and incubated with 20 ug/ml anti-LC3B (1251A) and 1:500 anti-tubulin NB100-690) for 1 h at room temperature. LC3 reactivity (green) was detected with ant-rabbit Dylight 488 and tubulin (red) with anti-mouse Dylight 550. Nuclei were counterstained with DAPI (blue).