Product datasheet
Anti-HECT E3 ubiquitin ligase antibody ab83853
3 Images
Overview
Product name
Anti-HECT E3 ubiquitin ligase antibody
Description
Rabbit polyclonal to HECT E3 ubiquitin ligase
Tested applications
ICC/IF, WB, IHC-P
Species reactivity
Reacts with: Human
Predicted to work with: Rabbit
Immunogen
Synthetic peptide within residues: FHPEELKDVI VGNTDYDWKT FEKNARYEPG
YNSSHPTIVM FWKAFHKLTL , corresponding to internal sequence amino acids 900-949 of
Human HECT E3 ubiquitin ligase (NP_057407).
Run BLAST with
Positive control
Run BLAST with
HepG2 cell lysate.
Properties
Form
Liquid
Storage instructions
Shipped at 4°C. Upon delivery aliquot and store at -20°C. Avoid repeated freeze / thaw cycles.
Storage buffer
Preservative: None
Constituents: 2% Sucrose, PBS
Purity
Immunogen affinity purified
Clonality
Polyclonal
Isotype
IgG
Applications
Our Abpromise guarantee covers the use of ab83853 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
ICC/IF
Use a concentration of 1 µg/ml.
WB
Use a concentration of 1 µg/ml. Detects a band of approximately 117 kDa
(predicted molecular weight: 117 kDa). Good results were obtained when
blocked with 5% non-fat dry milk in 0.05% PBS-T.
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Application
Abreviews
Notes
IHC-P
1/100.
Target
Function
Major E3 ligase for ISG15 conjugation. Acts as a positive regulator of innate antiviral response
in cells induced by interferon. Makes part of the ISGylation machinery that recognizes target
proteins in a broad and relatively non-specific manner. Catalyzes ISGylation of IRF3 which
results in sustained activation, it attenuates IRF3-PIN1 interaction, which antagonizes IRF3
ubiquitination and degradation, and boosts the antiviral response. Catalyzes ISGylation of
influenza A viral NS1 which attenuates virulence; ISGylated NS1 fails to form homodimers and
thus to interact with its RNA targets. Catalyzes ISGylation of papillomavirus type 16 L1 protein
which results in dominant-negative effect on virus infectivity. Physically associated with
polyribosomes, broadly modifies newly synthesized proteins in a cotranslational manner. In an
interferon-stimulated cell, newly translated viral proteins are primary targets of ISG15.
Tissue specificity
Expressed in testis and to a lesser degree in brain, ovary and placenta. Found in most tissues at
low levels.
Sequence similarities
Contains 1 HECT (E6AP-type E3 ubiquitin-protein ligase) domain.
Contains 5 RCC1 repeats.
Post-translational
modifications
ISGylated.
Cellular localization
Cytoplasm > perinuclear region. Associated with the polyribosomes, probably via the 60S
subunit.
Anti-HECT E3 ubiquitin ligase antibody images
Immunohistochemistry (Formalin/PFA-fixed
paraffin-embedded sections) analysis of
human pineal tissue labelling HECT E3
ubiquitin ligase with ab83853 at 1/100. A
Immunohistochemistry (Formalin/PFA-fixed
paraffin-embedded sections) - Anti-HECT E3
ubiquitin ligase antibody (ab83853)
Cy3-conjugated donkey anti-rabbit IgG
(1/200) was used as the secondary antibody.
Positive staining shown in the cytoplasm of
cell bodies and processes of pinealocytes.
Magnification: 20X. Exposure time: 0.5 - 2.0
seconds. Left - DAPI. Middle - HECT E3
ubiquitin ligase. Right - Merge.
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Anti-HECT E3 ubiquitin ligase antibody
(ab83853) at 1 µg/ml (5% skim milk / PBS ) +
HepG2 cell lysate at 10 µg
Secondary
HRP conjugated anti-Rabbit IgG at 1/50000
dilution
Predicted band size : 117 kDa
Observed band size : 117 kDa
Western blot - HECT E3 ubiquitin ligase antibody
(ab83853)
Additional bands at : 55 kDa. We are
unsure as to the identity of these extra
bands.Gel concentration: 6-18%
ICC/IF image of ab83853 stained HeLa cells.
The cells were 4% formaldehyde (10 min) and
then incubated in 1%BSA / 10% normal goat
serum / 0.3M glycine in 0.1% PBS-Tween for
1h to permeabilise the cells and block nonspecific protein-protein interactions. The cells
were then incubated with the antibody
(ab83853, 1µg/ml) overnight at +4°C. The
secondary antibody (green) was ab96899
Immunocytochemistry/ Immunofluorescence -
Dylight 488 goat anti-rabbit IgG (H+L) used at
Anti-HECT E3 ubiquitin ligase antibody (ab83853)
a 1/250 dilution for 1h. Alexa Fluor® 594
WGA was used to label plasma membranes
(red) at a 1/200 dilution for 1h. DAPI was
used to stain the cell nuclei (blue) at a
concentration of 1.43µM.
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