Chloroquine treated HeLa cells were subjected to SDS PAGE followed by western blot with 12135-1-AP (LC3 Antibody) at dilution of 1:1000 incubated at room temperature for 1.5 hours.
Chloroquine treated HeLa cells were subjected to SDS PAGE followed by western blot with 12135-1-AP (LC3 Antibody) at dilution of 1:1000 incubated at room temperature for 1.5 hours.
WB analysis of HEK-293 using 12135-1-AP
Chloroquine treated HEK-293 cells were subjected to SDS PAGE followed by western blot with 12135-1-AP (LC3 Antibody) at dilution of 1:1000 incubated at room temperature for 1.5 hours.
Chloroquine treated HEK-293 cells were subjected to SDS PAGE followed by western blot with 12135-1-AP (LC3 Antibody) at dilution of 1:1000 incubated at room temperature for 1.5 hours.
IHC staining of human breast cancer using 12135-1-AP
Immunohistochemical analysis of paraffin-embedded human breast cancer tissue slide using 12135-1-AP (LC3 antibody) at dilution of 1:200 (under 10x lens. Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0).
Immunohistochemical analysis of paraffin-embedded human breast cancer tissue slide using 12135-1-AP (LC3 antibody) at dilution of 1:200 (under 10x lens. Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0).
IHC staining of human breast cancer using 12135-1-AP
Immunohistochemical analysis of paraffin-embedded human breast cancer tissue slide using 12135-1-AP (LC3 antibody) at dilution of 1:200 (under 40x lens. Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0).
Immunohistochemical analysis of paraffin-embedded human breast cancer tissue slide using 12135-1-AP (LC3 antibody) at dilution of 1:200 (under 40x lens. Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0).
IHC staining of human liver cancer using 12135-1-AP
Immunohistochemical analysis of paraffin-embedded human liver cancer tissue slide using 12135-1-AP (LC3 antibody) at dilution of 1:200 (under 10x lens).
Immunohistochemical analysis of paraffin-embedded human liver cancer tissue slide using 12135-1-AP (LC3 antibody) at dilution of 1:200 (under 10x lens).
IHC staining of human liver cancer using 12135-1-AP
Immunohistochemical analysis of paraffin-embedded human liver cancer tissue slide using 12135-1-AP (LC3 antibody) at dilution of 1:200 (under 40x lens).
Immunohistochemical analysis of paraffin-embedded human liver cancer tissue slide using 12135-1-AP (LC3 antibody) at dilution of 1:200 (under 40x lens).
IF Staining of HEK-293 using 12135-1-AP
Immunofluorescent analysis of starvation treated HEK-293 cells using 12135-1-AP (LC3 Antibody) at dilution of 1:50 and Alexa Fluor 488-conjugated Goat Anti-Rabbit IgG(H+L). Cells were fixed with ethanol at -20 °C. 20 mM chloroquine was used to block the autophagy flux.
Immunofluorescent analysis of starvation treated HEK-293 cells using 12135-1-AP (LC3 Antibody) at dilution of 1:50 and Alexa Fluor 488-conjugated Goat Anti-Rabbit IgG(H+L). Cells were fixed with ethanol at -20 °C. 20 mM chloroquine was used to block the autophagy flux.
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Chloroquine treated HEK-293 cells, Chloroquine treated HeLa cells
Positive IHC detected in
human breast cancer tissue, human liver cancer tissue Note: suggested antigen retrieval with TE buffer pH 9.0; (*) Alternatively, antigen retrieval may be performed with citrate buffer pH 6.0
Positive IF/ICC detected in
Starvation treated HEK-293 cells
Recommended dilution
Application
Dilution
Western Blot (WB)
WB : 1:500-1:2000
Immunohistochemistry (IHC)
IHC : 1:50-1:500
Immunofluorescence (IF)/ICC
IF/ICC : 1:50-1:500
It is recommended that this reagent should be titrated in each testing system to obtain optimal results.
Sample-dependent, Check data in validation data gallery.
PBS with 0.02% sodium azide and 50% glycerol pH 7.3.
Storage Conditions
Store at -20°C. Stable for one year after shipment. Aliquoting is unnecessary for -20oC storage. 20ul sizes contain 0.1% BSA.
Background Information
LC3, also known as microtubule-associated protein 1 light chain 3 (MAP1LC3), is a mammalian homolog of the yeast Atg8p (autophagy-related 8 protein) that is involved in autophagy process. Three isoforms of LC3 exist as LC3A,LC3B, and LC3C. LC3 is first synthesized as pro-LC3, and undergoes C-terminal cleavage to yield a cytosolic precursor known as LC3-I. During autophagy LC3-I is subsequently conjugated to PE lipid as lipidated form: LC3-II. The lipidated form migrates faster on SDS-PAGE gel. The increased the level of LC3-II has been widely used as autophagy marker. This antibody recognizes both LC3-I (18-20 kDa) and LC3-II (14-16 kDa).
Chloroquine treated HeLa cells were subjected to SDS PAGE followed by western blot with 12135-1-AP (LC3 Antibody) at dilution of 1:1000 incubated at room temperature for 1.5 hours.
WB analysis of HEK-293 using 12135-1-AP
Chloroquine treated HEK-293 cells were subjected to SDS PAGE followed by western blot with 12135-1-AP (LC3 Antibody) at dilution of 1:1000 incubated at room temperature for 1.5 hours.
IHC Figures
IHC staining of human breast cancer using 12135-1-AP
Immunohistochemical analysis of paraffin-embedded human breast cancer tissue slide using 12135-1-AP (LC3 antibody) at dilution of 1:200 (under 10x lens. Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0).
IHC staining of human breast cancer using 12135-1-AP
Immunohistochemical analysis of paraffin-embedded human breast cancer tissue slide using 12135-1-AP (LC3 antibody) at dilution of 1:200 (under 40x lens. Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0).
IHC staining of human liver cancer using 12135-1-AP
Immunohistochemical analysis of paraffin-embedded human liver cancer tissue slide using 12135-1-AP (LC3 antibody) at dilution of 1:200 (under 10x lens).
IHC staining of human liver cancer using 12135-1-AP
Immunohistochemical analysis of paraffin-embedded human liver cancer tissue slide using 12135-1-AP (LC3 antibody) at dilution of 1:200 (under 40x lens).
IF Figures
IF Staining of HEK-293 using 12135-1-AP
Immunofluorescent analysis of starvation treated HEK-293 cells using 12135-1-AP (LC3 Antibody) at dilution of 1:50 and Alexa Fluor 488-conjugated Goat Anti-Rabbit IgG(H+L). Cells were fixed with ethanol at -20 °C. 20 mM chloroquine was used to block the autophagy flux.
The species listed in Tested Reactivity are in-house verified and applicable species. For unlisted species, please refer to the homology analysis of the immunogen sequence and related species. For rabbit polyclonal antibodies, homology >70% is recommended. For mouse monoclonal antibodies and rabbit recombinant antibodies, homology >90% is recommended. Generally, the higher the homology, the greater the applicability. However, there will be certain differences in protein expression in different species, tissues or cells. Therefore, the homology analysis results are for reference only and do not serve as a guarantee.
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