Key features and details
- Mouse monoclonal [31A3] to PGP9.5
- Suitable for: Flow Cyt, IHC-P, WB
- Knockout validated
- Reacts with: Rat, Human
- Isotype: IgG1
Product nameAnti-PGP9.5 antibody [31A3]
See all PGP9.5 primary antibodies
DescriptionMouse monoclonal [31A3] to PGP9.5
Tested applicationsSuitable for: Flow Cyt, IHC-P, WBmore details
Species reactivityReacts with: Rat, Human
Full length protein corresponding to PGP9.5.
This antibody clone is manufactured by Abcam.
This monoclonal antibody to PGP9.5 has been knockout validated in Western blot. The expected band for PGP9.5 was observed in wild type cells and the band was not seen in knockout cells.
Storage instructionsShipped at 4°C. Upon delivery aliquot and store at -20°C. Avoid freeze / thaw cycles.
Storage bufferpH: 7.20
Preservative: 0.02% Sodium azide
Constituents: PBS, 6.97% L-Arginine
Concentration information loading...
PurityProtein A purified
Our Abpromise guarantee covers the use of ab20559 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|Flow Cyt||Use 0.1-1µg for 106 cells.
ab170190 - Mouse monoclonal IgG1, is suitable for use as an isotype control with this antibody.
|IHC-P||Use at an assay dependent concentration.|
|WB||Use a concentration of 1 - 5 µg/ml. Detects a band of approximately 25 kDa (predicted molecular weight: 25 kDa).
Dilution optimised using Chromogenic detection.
FunctionUbiquitin-protein hydrolase involved both in the processing of ubiquitin precursors and of ubiquitinated proteins. This enzyme is a thiol protease that recognizes and hydrolyzes a peptide bond at the C-terminal glycine of ubiquitin. Also binds to free monoubiquitin and may prevent its degradation in lysosomes. The homodimer may have ATP-independent ubiquitin ligase activity.
Tissue specificityFound in neuronal cell bodies and processes throughout the neocortex (at protein level). Expressed in neurons and cells of the diffuse neuroendocrine system and their tumors. Weakly expressed in ovary. Down-regulated in brains from Parkinson disease and Alzheimer disease patients.
Involvement in diseaseParkinson disease 5
Neurodegeneration with optic atrophy, childhood-onset
Sequence similaritiesBelongs to the peptidase C12 family.
Cellular localizationCytoplasm. Endoplasmic reticulum membrane. About 30% of total UCHL1 is associated with membranes in brain.
- Information by UniProt
- Epididymis luminal protein 117 antibody
- Epididymis secretory protein Li 53 antibody
- HEL 117 antibody
Immunohistochemical analysis of rat emybronic rectal tissue, staining PGP9.5 with ab20559.
After antigen retrieval, the sections were treated and incubated with primary antibody (1/200), followed by an HRP-conjugated donkey anti-mouse IgG (1/200). Staining was detected using DAB.
Lane 1: Wild type HAP1 whole cell lysate (20 µg)
Lane 2: UCHL1 (KO) knockout HAP1 whole cell lysate (20 µg)
Lane 3: SH-SY5Y whole cell lysate (20 µg)
Lane 4: Hek293 whole cell lysate (20 µg)
Lanes 1 - 4: Merged signal (red and green). Green - ab20559 observed at 24 kDa. Red - loading control, ab181602, observed at 37 kDa.
ab20559 detected the expected band for PGP9.5 in wild type cells and the band was not seen in PGP9.5/UCHL1 knockout cells. Wild-type and UCHL1 knockout samples were subjected to SDS-PAGE. Ab20559 and ab181602 (Rabbit anti GAPDH loading control) were incubated overnight at 4°C at 1 ug/ml and 1/10000 dilution respectively. Blots were developed with Goat anti-Mouse IgG H&L (IRDye® 800CW) preabsorbed ab216772 and Goat anti-Rabbit IgG H&L (IRDye® 680RD) preabsorbed ab216777 secondary antibodies at 1/10000 dilution for 1 hour at room temperature before imaging.
Overlay histogram showing SH-SH5Y cells stained with ab20559 (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 (ab20559, 0.1μg/1x106 cells) for 30 min at 22°C. The secondary antibody used was Alexa Fluor® 488 goat anti-mouse IgG (H&L) (ab150113) at 1/2000 dilution for 30 min at 22°C. Isotype control antibody (black line) was mouse IgG1 [ICIGG1] (ab91353, 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 SH-SY5Y cells fixed with 4% paraformaldehyde (10 min)/permeabilized with 0.1% PBS-Tween for 20 min used under the same conditions.
All lanes : Anti-PGP9.5 antibody [31A3] (ab20559) at 5 µg/ml
Lane 1 : Human brain tissue lysate - total protein (ab29466)
Lane 2 : Human spinal cord tissue lysate - total protein (ab29188)
Lane 3 : Brain (Rat) Tissue Lysate
Lane 4 : Brain (Mouse) Tissue Lysate
Lane 5 : SHSY-5Y (Human neuroblastoma cell line) Whole Cell Lysate
Lysates/proteins at 20 µg per lane.
All lanes : Goat Anti-Mouse IgG H&L (HRP) preadsorbed (ab97040) at 1/50000 dilution
Developed using the ECL technique.
Performed under reducing conditions.
Predicted band size: 25 kDa
Additional bands at: 25 kDa. We are unsure as to the identity of these extra bands.
Exposure time: 4 minutes
ab20559 has been referenced in 8 publications.
- Mtango NR et al. Essential role of maternal UCHL1 and UCHL3 in fertilization and preimplantation embryo development. J Cell Physiol 227:1592-603 (2012). ICC/IF ; Mouse . PubMed: 21678411
- Saygili E et al. Age-related regional differences in cardiac nerve growth factor expression. Age (Dordr) 34:659-67 (2012). WB ; Rat . PubMed: 21559866
- Mtango NR et al. Essential role of ubiquitin C-terminal hydrolases UCHL1 and UCHL3 in mammalian oocyte maturation. J Cell Physiol 227:2022-9 (2012). PubMed: 21751213
- Wang W et al. Abnormal innervation patterns in the anorectum of ETU-induced fetal rats with anorectal malformations. Neurosci Lett 495:88-92 (2011). IHC-P ; Rat . PubMed: 21440597
- Zippin JH et al. Soluble adenylyl cyclase defines a nuclear cAMP microdomain in keratinocyte hyperproliferative skin diseases. J Invest Dermatol 130:1279-87 (2010). IHC-P ; Human . PubMed: 20130594
- Rana OR et al. Acetylcholine as an age-dependent non-neuronal source in the heart. Auton Neurosci 156:82-9 (2010). WB ; Rat . PubMed: 20510655
- Crimmins S et al. Transgenic rescue of ataxia mice reveals a male-specific sterility defect. Dev Biol 325:33-42 (2009). IHC-P ; Mouse . PubMed: 18926813
- Yi YJ et al. Ubiquitin C-terminal hydrolase-activity is involved in sperm acrosomal function and anti-polyspermy defense during porcine fertilization. Biol Reprod 77:780-93 (2007). IHC-P ; Mouse . PubMed: 17671268