Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR21235] to LC3B
- Suitable for: ICC/IF, IHC-P
- Knockout validated
- Reacts with: Human
Product nameAnti-LC3B antibody [EPR21235]
See all LC3B primary antibodies
DescriptionRabbit monoclonal [EPR21235] to LC3B
Tested applicationsSuitable for: ICC/IF, IHC-Pmore details
Species reactivityReacts with: Human
Recombinant full length protein.
Database link: Q9GZQ8
- ICC/IF: HAP1 cells (+/- chloroquine) (HAP1-MAP1LC3B knockout cells used as negative cell line). IHC-P: FFPE human normal colon and human normal cerebral cortex tissue sections.
This product was made using synthetic libraries and phage display technology.
This antibody is a recombinant chimeric antibody. Rabbit chimeric monoclonal antibody (Human Fab/ Rabbit Fc).
Storage instructionsShipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C long term. Avoid freeze / thaw cycle.
Storage bufferPreservative: 0.02% Sodium azide
Constituents: 1% BSA, PBS
Concentration information loading...
PurityProtein A purified
- Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077)
- Goat Anti-Rabbit IgG H&L (Alexa Fluor® 555) (ab150078)
- Goat Anti-Rabbit IgG H&L (Alexa Fluor® 647) (ab150079)
- Goat Anti-Rabbit IgG H&L (Alexa Fluor® 594) (ab150080)
- Goat Anti-Rabbit IgG H&L (HRP) (ab205718)
- Goat Anti-Rabbit IgG H&L (DyLight® 488) preadsorbed (ab96899)
- Goat Anti-Rabbit IgG H&L (HRP) (ab97051)
Our Abpromise guarantee covers the use of ab239416 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|ICC/IF||Use a concentration of 0.1 µg/ml.|
|IHC-P||Use a concentration of 5 µg/ml. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.|
FunctionProbably involved in formation of autophagosomal vacuoles (autophagosomes).
Tissue specificityMost abundant in heart, brain, skeletal muscle and testis. Little expression observed in liver.
Sequence similaritiesBelongs to the MAP1 LC3 family.
modificationsThe precursor molecule is cleaved by APG4B/ATG4B to form LC3-I. This is activated by APG7L/ATG7, transferred to ATG3 and conjugated to phospholipid to form LC3-II.
Cellular localizationCytoplasm > cytoskeleton. Endomembrane system. Cytoplasmic vesicle > autophagosome membrane. LC3-II binds to the autophagic membranes.
- Information by UniProt
- ATG8F antibody
- Autophagy-related protein LC3 B antibody
- Autophagy-related ubiquitin-like modifier LC3 B antibody
ab239416 staining LC3B in HAP1 cells (wildtype and MAP1LC3B knockout) +/- Chloroquine (50μM, 24 hours). The cells were fixed with 100% methanol (5 min), permeabilized with 0.1% 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 ab239416 at 0.1μg/ml and ab195889 then detected with an Alexa Fluor® 488 goat anti-rabbit secondary antibody (ab150081) at a 1/1000 dilution (shown in green). Nuclear DNA was labelled with DAPI (shown in blue), and ab195889, Mouse monoclonal to alpha Tubulin (Alexa Fluor® 594), at a 1/250 dilution (shown in red).
IHC image of LC3B staining in a section of formalin-fixed paraffin-embedded normal human cerebral cortex performed on a Leica BONDTM system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20mins. The section was then incubated with ab239416, 5ug/ml, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX. The inset secondary-only control image is taken from an identical assay without primary antibody.
For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.
To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.
ab239416 has not yet been referenced specifically in any publications.