Storage instructionsShipped at 4°C. Upon delivery aliquot and store at -20°C or -80°C. Avoid repeated freeze / thaw cycles.
Storage bufferPreservative: None
PBS, pH 7.2
Concentration information loading...
PurityProtein G purified
Light chain typekappa
Our Abpromise guarantee covers the use of ab55474 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|WB||Use a concentration of 1 - 5 µg/ml.|
|IP||Use at an assay dependent concentration. PubMed: 23626693|
|IHC-P||Use a concentration of 4 µg/ml.|
|Flow Cyt||Use 1µg for 106 cells.
ab170191 - Mouse monoclonal IgG2a, is suitable for use as an isotype control with this antibody.
FunctionActs probably as a receptor for selective autophagosomal degradation of ubiquitinated targets.
Sequence similaritiesContains 1 OPR domain.
Contains 1 UBA domain.
Contains 1 ZZ-type zinc finger.
DomainThe OPR domain mediates interaction with SQSTM1.
Cellular localizationCytoplasm. Cytoplasmic vesicle > autophagosome. Lysosome. Cytoplasm > myofibril > sarcomere > M line. In cardiac muscles localizes to the sarcomeric M line (By similarity). Is targeted to lysosomes for degradation.
- Information by UniProt
- 1A1 3B antibody
- 1A13B antibody
- B box protein antibody
Western blot against tagged recombinant protein immunogen using ab55474 NBR1 antibody at 1ug/ml. Predicted band size of immunogen is 37 kDa.
This antibody has only been tested in WB against the recombinant fragment used as immunogen. We have no data on the detection of endogenous protein.
Overlay histogram showing HeLa cells stained with ab55474 (red line). The cells were fixed with 80% methanol (5 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab55474, 1μg/1x106 cells) for 30 min at 22°C. The secondary antibody used was DyLight® 488 goat anti-mouse IgG (H+L) (ab96879) at 1/500 dilution for 30 min at 22°C. Isotype control antibody (black line) was mouse IgG2a [ICIGG2A] (ab91361, 1μg/1x106 cells) used under the same conditions. Unlabelled sample (blue line) was also used as a control. Acquisition of >5,000 events were collected using a 20mW Argon ion laser (488nm) and 525/30 bandpass filter. This antibody gave a positive signal in HeLa cells fixed with 4% paraformaldehyde (10 min)/permeabilized with 0.1% PBS-Tween for 20 min used under the same conditions.
ab55474 (4µg/ml) staining NBR1 in human skeletal muscle using an automated system (DAKO Autostainer Plus). Using this protocol there is moderate cytoplasmic staining.
Sections were rehydrated and antigen retrieved with the Dako 3 in 1 AR buffers EDTA pH 9.0 in a DAKO PT link. Slides were peroxidase blocked in 3% H2O2 in methanol for 10 mins. They were then blocked with Dako Protein block for 10 minutes (containing casein 0.25% in PBS) then incubated with primary antibody for 20 min and detected with Dako envision flex amplification kit for 30 minutes. Colorimetric detection was completed with Diaminobenzidine for 5 minutes. Slides were counterstained with Haematoxylin and coverslipped under DePeX. Please note that, for manual staining, optimization of primary antibody concentration and incubation time is recommended. Signal amplification may be required.
This product has been referenced in:
- Yu Z et al. Autophagy defects and related genetic variations in renal cell carcinoma with eosinophilic cytoplasmic inclusions. Sci Rep 8:9972 (2018). Read more (PubMed: 29967346) »
- Lin X et al. Liver-specific deletion of Eva1a/Tmem166 aggravates acute liver injury by impairing autophagy. Cell Death Dis 9:768 (2018). Read more (PubMed: 29991758) »