Anti-LC3B antibody [EPR18709] - Autophagosome Marker – Chicken IgY (Chimeric)

4 product images

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  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-LC3B antibody [EPR18709] - Autophagosome Marker – Chicken IgY (Chimeric) (ab322912)

  Immunofluorescence staining of LC3B in sections of formalin-fixed paraffin-embedded mouse brain (positive) and mouse liver (negative).
  Performed on a Leica BOND. The section was pre-treated using heat mediated antigen retrieval with sodium citrate (pH6.0) for 20 minutes. The section was then incubated at room temperature for 1 hour with ab322912 at 1/100 dilution and then incubated for 1 hour with Goat Anti-Chicken IgY H&L (Alexa Fluor® 488) preadsorbed ab150173 Goat Anti-Chicken IgY H&L (Alexa Fluor® 488) preadsorbed at 1/1000 dilution (shown in green). Nuclear DNA was labelled with DAPI (shown in blue). The section was then mounted using Dako Fluorescence Mounting Medium®.
  Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).
  For other IHC staining systems (automated and non-automated), customers should optimize variable parameters such as antigen retrieval conditions, antibody concentrations and incubation times.

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  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-LC3B antibody [EPR18709] - Autophagosome Marker – Chicken IgY (Chimeric) (ab322912)

  Immunofluorescence staining of LC3B in sections of formalin-fixed paraffin-embedded human brain (positive) and human heart muscle (negative)*.
  Performed on a Leica BOND. The section was pre-treated using heat mediated antigen retrieval with sodium citrate (pH6.0) for 20 minutes. The section was then incubated at room temperature for 1 hour with ab322912 at 1/100 dilution and then incubated for 1 hour with Goat Anti-Chicken IgY H&L (Alexa Fluor® 488) preadsorbed ab150173 Goat Anti-Chicken IgY H&L (Alexa Fluor® 488) preadsorbed at 1/1000 dilution (shown in green). Nuclear DNA was labelled with DAPI (shown in blue). The section was then mounted using Dako Fluorescence Mounting Medium®.
  Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).
  For other IHC staining systems (automated and non-automated), customers should optimize variable parameters such as antigen retrieval conditions, antibody concentrations and incubation times.
  *Tissue obtained from the Human Research Tissue Bank, supported by the NIHR Cambridge Biomedical Research Centre.

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  Immunocytochemistry/ Immunofluorescence - Anti-LC3B antibody [EPR18709] - Autophagosome Marker – Chicken IgY (Chimeric) (ab322912)

  ab322912 staining LC3B in NIH/3T3 +/- Chloroquine (50µM 24 hours) cells. The cells were fixed with 100% methanol (5 min), permeabilized with 0.1% PBS-Triton X-100 for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1%PBS-Tween for 1h. The cells were then incubated overnight at 4°C with ab322912 at 1µg/ml (shown in green) and Alexa Fluor® 594 Anti-alpha Tubulin antibody [EP1332Y] - Microtubule Marker ab202272, Alexa Fluor® 594 Rabbit monoclonal [EP1332Y] to alpha Tubulin - Microtubule Marker (shown in pseudocolour magenta). Cells were then incubated with Goat Anti-Chicken IgY H&L (Alexa Fluor® 488) preadsorbed ab150173, Goat Anti-Chicken IgY H&L (Alexa Fluor® 488) preadsorbed at 1/1000 dilution. Nuclear DNA was labelled with DAPI (shown in blue). Image was acquired with a high-content analyser (Operetta CLS, Perkin Elmer) and a maximum intensity projection of confocal sections is shown.

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  Immunocytochemistry/ Immunofluorescence - Anti-LC3B antibody [EPR18709] - Autophagosome Marker – Chicken IgY (Chimeric) (ab322912)

  ab322912 staining LC3B in HAP1 cells (wildtype and MAP1LC3B knockout) +/- Chloroquine (50µM 24 hours) cells. The cells were fixed with 100% methanol (5 min), permeabilized with 0.1% PBS-Triton X-100 for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1%PBS-Tween for 1h. The cells were then incubated overnight at 4°C with ab322912 at 1µg/ml (shown in green) and Alexa Fluor® 594 Anti-alpha Tubulin antibody [EP1332Y] - Microtubule Marker ab202272, Alexa Fluor® 594 Rabbit monoclonal [EP1332Y] to alpha Tubulin - Microtubule Marker (shown in pseudocolour magenta). Cells were then incubated with Goat Anti-Chicken IgY H&L (Alexa Fluor® 488) preadsorbed ab150173, Goat Anti-Chicken IgY H&L (Alexa Fluor® 488) preadsorbed at 1/1000 dilution. Nuclear DNA was labelled with DAPI (shown in blue). Image was acquired with a high-content analyser (Operetta CLS, Perkin Elmer) and a maximum intensity projection of confocal sections is shown.