Product nameAnti-PGP9.5 antibody [346CT2.5.1]
See all PGP9.5 primary antibodies
DescriptionMouse monoclonal [346CT2.5.1] to PGP9.5
Tested applicationsSuitable for: IHC-P, Flow Cyt, WBmore details
Species reactivityReacts with: Rat, Goat, Cat, Dog, Human
Predicted to work with: Mouse, Horse, Pig
Recombinant full length protein corresponding to Human PGP9.5 aa 1 to the C-terminus.
- This antibody gave a positive signal in the following lysates: Human Brain Tissue; Rat Brain Tissue; Rat Brain Cortex Tissue; PC12 Whole Cell; Y79 Whole Cell; U-87MG Whole Cell; SHSY-5Y Whole Cell and HAP1 Wole cells.
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: 7.40
Preservative: 0.02% Sodium azide
Concentration information loading...
PurityProtein G purified
Our Abpromise guarantee covers the use of ab86808 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|IHC-P||1/1000. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.|
|Flow Cyt||Use 0.01-0.1µg for 106 cells.
ab170190 - Mouse monoclonal IgG1, is suitable for use as an isotype control with this antibody.
|WB||Use a concentration of 1 µg/ml. Detects a band of approximately 25 kDa (predicted molecular weight: 25 kDa).
Abcam recommends blocking with 3% Milk.
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
IHC-P image of PGP9.5 staining on cat bladder sections using ab86808 (1:3000). The sections were subjected to heat mediated antigen retrieval using citric acid. The sections were then blocked using 1% BSA at 21°C for 10 min. ab86808 was incubated at 21°C for 1 hours. The secondary antibody was Goat polyclonal to anti-mouse (rat pre-absorbed) conjugated to biotin (1:200).
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)
Lanes 1 - 3: Merged signal (red and green). Green - ab86808 observed at 24 kDa. Red - loading control, ab181602, observed at 37 kDa.
Ab86808 was shown to specifically react with PGP9.5/UCHL1 in wild-type cells as signal was lost in PGP9/UCHL1 knockout HAP1 cells. Wild-type and UCHL1 (KO) knockout samples were subjected to SDS-PAGE. Ab86808 and ab181602 (Rabbit anti GAPDH loading control) were incubated overnight at 4°C at 1 ug/ml and 1/2000 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 ab86808 (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 (ab86808, 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.
IHC-P image of PGP9.5 staining using ab86808(1:2500) on dog small intestine sections. The sections were subjected to heat mediated antigen retrieval using citric acid. The sections were then blocked with 1% BSA for 10 min at 21°C. the sections were then incubated with primary antibody ab86808 at 1:2500 dlution for 2 hours at 21°C. The secondary used was Goat polyclonal to anti mouse IgG conjugated to biotin (1:200)
All lanes : Anti-PGP9.5 antibody [346CT2.5.1] (ab86808) at 1 µg/ml
Lane 1 : Human brain tissue lysate - total protein (ab29466)
Lane 2 : Brain (Rat) Tissue Lysate
Lane 3 : Rat Cortex Tissue Lysate
Lane 4 : PC12 (Rat adrenal pheochromocytoma cell line) Whole Cell Lysate
Lane 5 : Y79 (Human retinoblastoma cell line) Whole Cell Lysate
Lane 6 : U-87 MG (Human glioblastoma astrocytoma) Whole Cell Lysate
Lane 7 : 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 1/5000 dilution
Developed using the ECL technique.
Performed under reducing conditions.
Predicted band size: 25 kDa
Observed band size: 25 kDa
Exposure time: 1 minute
IHC-P image of PGP9.5 staining on rat pancreas using ab86808 (1:1000). The rat pancreatic sections were subjected to heat mediated antigen retrieval using citric acid. The sections were then blocked using 1% BSA at 21°C for 10 min. ab86808 was incubated at 21°C for 1 hours. The secondary antibody was Goat polyclonal to anti-mouse (rat pre-absorbed) conjugated to biotin (1:200).
This product has been referenced in:
- Bautzova T et al. 5-oxoETE triggers nociception in constipation-predominant irritable bowel syndrome through MAS-related G protein-coupled receptor D. Sci Signal 11:N/A (2018). Read more (PubMed: 30563864) »
- Cenac N et al. Quantification and Potential Functions of Endogenous Agonists of Transient Receptor Potential Channels in Patients With Irritable Bowel Syndrome. Gastroenterology 149:433-44.e7 (2015). Read more (PubMed: 25911511) »