The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
Application notesFlow Cyt: 1/50.
ICC: 1/50 - 1/100.
IHC-P: 1/100 - 1/250. Perform heat mediated antigen retrieval before commencing with IHC staining protocol. Use of HRP-conjugated or polymerized HRP secondary antibodies recommended, stronger signals have been found using the polymerized HRP secondary.
WB: 1/10000 - 1/20000. Detects a band of approximately 24 kDa (predicted molecular weight: 20 kDa).
Is unsuitable for IP.
Not yet tested in other applications.
Optimal dilutions/concentrations should be determined by the end user.
FunctionProtects cells against oxidative stress and cell death. Plays a role in regulating expression or stability of the mitochondrial uncoupling proteins SLC25A14 and SLC25A27 in dopaminergic neurons of the substantia nigra pars compacta and attenuates the oxidative stress induced by calcium entry into the neurons via L-type channels during pacemaking. Eliminates hydrogen peroxide and protects cells against hydrogen peroxide-induced cell death. May act as an atypical peroxiredoxin-like peroxidase that scavenges hydrogen peroxide. Following removal of a C-terminal peptide, displays protease activity and enhanced cytoprotective action against oxidative stress-induced apoptosis. Stabilizes NFE2L2 by preventing its association with KEAP1 and its subsequent ubiquitination. Binds to OTUD7B and inhibits its deubiquitinating activity. Enhances RELA nuclear translocation. Binds to a number of mRNAs containing multiple copies of GG or CC motifs and partially inhibits their translation but dissociates following oxidative stress. Required for correct mitochondrial morphology and function and for autophagy of dysfunctional mitochondria. Regulates astrocyte inflammatory responses. Acts as a positive regulator of androgen receptor-dependent transcription. Prevents aggregation of SNCA. Plays a role in fertilization. Has no proteolytic activity. Has cell-growth promoting activity and transforming activity. May function as a redox-sensitive chaperone.
Tissue specificityHighly expressed in pancreas, kidney, skeletal muscle, liver, testis and heart. Detected at slightly lower levels in placenta and brain. Detected in astrocytes, Sertoli cells, spermatogonia, spermatids and spermatozoa.
Involvement in diseaseDefects in PARK7 are the cause of Parkinson disease type 7 (PARK7) [MIM:606324]. A neurodegenerative disorder characterized by resting tremor, postural tremor, bradykinesia, muscular rigidity, anxiety and psychotic episodes. PARK7 has onset before 40 years, slow progression and initial good response to levodopa. Some patients may show traits reminiscent of amyotrophic lateral sclerosis-parkinsonism/dementia complex (Guam disease).
Sequence similaritiesBelongs to the peptidase C56 family.
Post-translational modificationsSumoylated on Lys-130 by PIAS2 or PIAS4; which is enhanced after ultraviolet irradiation and essential for cell-growth promoting activity and transforming activity. Cys-106 is easily oxidized to sulfinic acid. Undergoes cleavage of a C-terminal peptide and subsequent activation of protease activity in response to oxidative stress.
Cellular localizationCytoplasm. Nucleus. Mitochondrion. Under normal conditions, located predominantly in the cytoplasm and, to a lesser extent, in the nucleus and mitochondrion. Translocates to the mitochondrion and subsequently to the nucleus in response to oxidative stress and exerts an increased cytoprotective effect against oxidative damage. Detected in tau inclusions in brains from neurodegenerative disease patients.
Overlay histogram showing Jurkat cells stained with ab76241 (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 (ab76241, 1/100 dilution) for 30 min at 22°C. The secondary antibody used was Alexa Fluor® 488 goat anti-rabbit IgG (H+L) (ab150077) at 1/2000 dilution for 30 min at 22°C. Isotype control antibody (black line) was rabbit IgG (monoclonal) (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 Jurkat cells fixed with 4% paraformaldehyde (10 min)/permeabilized with 0.1% PBS-Tween for 20 min used under the same conditions.
Western blot - DJ1 antibody [EP2816Y] (ab76241)
All lanes : Anti-PARK7/DJ1 antibody [EP2816Y] (ab76241) at 1/20000 dilution
Lane 1 : TF-1 cell lysate Lane 2 : Jurkat cell lysate Lane 3 : HeLa cell lysate
Lysates/proteins at 10 µg per lane.
Secondary HRP labelled goat anti-rabbit at 1/1000 dilution
Immunohistochemical analysis of paraffin-embedded human brain tissue using ab76241 at 1/100 dilution.
References for Anti-PARK7/DJ1 antibody [EP2816Y] (ab76241)
This product has been referenced in:
Prahlad J et al. Use of cysteine-reactive cross-linkers to probe conformational flexibility of human DJ-1 demonstrates that Glu18 mutations are dimers. J Neurochem130:839-53 (2014).
Read more (PubMed: 24832775) »
Besong Agbo D et al. Development of a capillary isoelectric focusing immunoassay to measure DJ-1 isoforms in biological samples. Anal Biochem443:197-204 (2013).
Read more (PubMed: 24055619) »