Product datasheet
Anti-NBR1 antibody ab55474
1 Abreviews 5 References 3 Images
Overview
Product name
Anti-NBR1 antibody
Description
Mouse monoclonal to NBR1
Tested applications
WB, IP, IHC-P, Flow Cyt
Species reactivity
Reacts with: Mouse, Human
Immunogen
Recombinant fragment: EPQVTLNVTF KNEIQSFLVS DPENTTWADI EAMVKVSFDL
NTIQIKYLDE ENEEVSINSQ GEYEEALKMA VKQGNQLQMQ VHEGHHVVDE APPPV,
corresponding to amino acids 2-97 of Human NBR1
Run BLAST with
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Properties
Form
Liquid
Storage instructions
Shipped at 4°C. Upon delivery aliquot and store at -20°C or -80°C. Avoid repeated freeze / thaw
cycles.
Storage buffer
Preservative: None
PBS, pH 7.2
Purity
Protein G purified
Clonality
Monoclonal
Isotype
IgG2a
Light chain type
kappa
Applications
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.
Application
Abreviews
Notes
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.
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Application
Abreviews
Notes
Use 1µg for 106 cells. ab170191-Mouse monoclonal IgG2a, is suitable for use as
Flow Cyt
an isotype control with this antibody.
Target
Function
Acts probably as a receptor for selective autophagosomal degradation of ubiquitinated targets.
Sequence similarities
Contains 1 OPR domain.
Contains 1 UBA domain.
Contains 1 ZZ-type zinc finger.
Domain
The OPR domain mediates interaction with SQSTM1.
Cellular localization
Cytoplasm. 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.
Anti-NBR1 antibody images
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
Flow Cytometry - Anti-NBR1 antibody (ab55474)
(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.
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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.
Western blot - NBR1 antibody (ab55474)
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
Immunohistochemistry (Formalin/PFA-fixed
paraffin-embedded sections) - NBR1 antibody
(ab55474)
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.
Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"
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