Anti-PGP9.5 antibody (ab27053)
Key features and details
- Rabbit polyclonal to PGP9.5
- Suitable for: IHC-P, IP, WB, ICC/IF
- Knockout validated
- Reacts with: Mouse, Rat, Human
- Isotype: IgG
Get better batch-to-batch reproducibility with a recombinant antibody
- Research with confidence – consistent and reproducible results with every batch
- Long-term and scalable supply – powered by recombinant technology for fast production
- Success from the first experiment – confirmed specificity through extensive validation
- Ethical standards compliant – production is animal-free
Overview
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Product name
Anti-PGP9.5 antibody
See all PGP9.5 primary antibodies -
Description
Rabbit polyclonal to PGP9.5 -
Host species
Rabbit -
Tested applications
Suitable for: IHC-P, IP, WB, ICC/IFmore details -
Species reactivity
Reacts with: Mouse, Rat, Human -
Immunogen
Synthetic peptide corresponding to Human PGP9.5 aa 150 to the C-terminus conjugated to keyhole limpet haemocyanin.
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Positive control
- ICC/IF: PC12 cells and primary rat neurons/glia, DIV14 cells. WB: Wild type HAP1 whole cell lysate. HEK-293 whole cell lysate. Human, rat and mouse brain whole cell lysate. IHC-P: Human pancreas tissue. IP: Mouse brain tissue lysate.
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General notes
The Life Science industry has been in the grips of a reproducibility crisis for a number of years. Abcam is leading the way in addressing this with our range of recombinant monoclonal antibodies and knockout edited cell lines for gold-standard validation. Please check that this product meets your needs before purchasing.
If you have any questions, special requirements or concerns, please send us an inquiry and/or contact our Support team ahead of purchase. Recommended alternatives for this product can be found below, along with publications, customer reviews and Q&As
Properties
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Form
Liquid -
Storage instructions
Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C or -80°C. Avoid freeze / thaw cycle. -
Storage buffer
pH: 7.40
Preservative: 0.02% Sodium azide
Constituent: PBS
Batches of this product that have a concentration < 1mg/ml may have BSA added as a stabilising agent. If you would like information about the formulation of a specific lot, please contact our scientific support team who will be happy to help. -
Concentration information loading...
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Purity
Immunogen affinity purified -
Clonality
Polyclonal -
Isotype
IgG -
Research areas
Associated products
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Compatible Secondaries
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Isotype control
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Positive Controls
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Recombinant Protein
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab27053 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 |
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IHC-P | (3) |
Use a concentration of 1 µg/ml. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
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IP |
Use a concentration of 5 µg/ml.
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WB | (1) |
Use a concentration of 1 µg/ml. Detects a band of approximately 25 kDa (predicted molecular weight: 25 kDa).
Abcam recommends using milk as the blocking agent. Abcam welcomes customer feedback and would appreciate any comments regarding this product and the data presented below. |
ICC/IF | (1) |
Use a concentration of 1 µg/ml.
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Notes |
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IHC-P
Use a concentration of 1 µg/ml. Perform heat mediated antigen retrieval before commencing with IHC staining protocol. |
IP
Use a concentration of 5 µg/ml. |
WB
Use a concentration of 1 µg/ml. Detects a band of approximately 25 kDa (predicted molecular weight: 25 kDa). Abcam recommends using milk as the blocking agent. Abcam welcomes customer feedback and would appreciate any comments regarding this product and the data presented below. |
ICC/IF
Use a concentration of 1 µg/ml. |
Target
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Function
Ubiquitin-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 specificity
Found 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 disease
Parkinson disease 5
Neurodegeneration with optic atrophy, childhood-onset -
Sequence similarities
Belongs to the peptidase C12 family. -
Post-translational
modificationsO-glycosylated. -
Cellular localization
Cytoplasm. Endoplasmic reticulum membrane. About 30% of total UCHL1 is associated with membranes in brain. - Information by UniProt
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Database links
- Entrez Gene: 7345 Human
- Entrez Gene: 22223 Mouse
- Entrez Gene: 29545 Rat
- Omim: 191342 Human
- SwissProt: P09936 Human
- SwissProt: Q9R0P9 Mouse
- SwissProt: Q00981 Rat
- Unigene: 518731 Human
see all -
Alternative names
- Epididymis luminal protein 117 antibody
- Epididymis secretory protein Li 53 antibody
- HEL 117 antibody
see all
Images
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Lane 1: Wild type HAP1 whole cell lysate (20 µg)
Lane 2: UCHL1 (PGP9.5) knockout HAP1 whole cell lysate (20 µg)
Lane 3: Hek293 whole cell lysate (20 µg)
Lane 4: Ms brain whole cell lysate (20 µg)Lanes 1 - 4: Merged signal (red and green). Green - ab27053 observed at 24 kDa. Red - loading control, ab8245, observed at 37 kDa.
ab27053 was shown to recognize UCHL1 (PGP9.5) in wild type cells as signal was lost at the expected MW in UCHL1 (PGP9.5) knockout cells. Additional cross-reactive bands were observed in the wild-type and knockout cells. Wild-type and UCHL1 (PGP9.5) knockout samples were subjected to SDS-PAGE. Ab27053 and ab8245 (Mouse 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-Rabbit IgG H&L (IRDye® 800CW) preabsorbed ab216773 and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed ab216776 secondary antibodies at 1/10000 dilution for 1 hour at room temperature before imaging.
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ab27053 staining PGP9.5 in PC12 cells. The cells were fixed with 100% methanol (5 min), permeabilized with 0.1% PBS-Tween for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1%PBS-Tween for 1h. The cells were then incubated overnight at 4°C with ab27053 at 1µg/ml and ab7291, Mouse monoclonal [DM1A] to alpha Tubulin - Loading Control. Cells were then incubated with ab150081, Goat polyclonal Secondary Antibody to Rabbit IgG - H&L (Alexa Fluor® 488), pre-adsorbed at 1/1000 dilution (shown in green) and ab150120, Goat polyclonal Secondary Antibody to Mouse IgG - H&L (Alexa Fluor® 594), pre-adsorbed at 1/1000 dilution (shown in pseudocolour red). Nuclear DNA was labelled with DAPI (shown in blue).
Image was acquired with a confocal microscope (Leica-Microsystems TCS SP8) and a single confocal section is shown.
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PGP9.5 antibody (ab27053)
IHC image of Anti-PGP9.5 antibody staining in a section of formalin-fixed paraffin-embedded normal human pancreas performed on a Leica BOND™ system using the standard protocol. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 minutes. The section was then incubated with ab27053, 1 µg/mL, for 15 minutes at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX.
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ab27053 staining PGP9.5 in primary rat neurons/glia, DIV14 (prepared from E18 rat hippocampal brain area, obtained from Transnetyx Tissue by BrainBits, LLC, cat.no. SDHEP) cells. The cells were fixed with 4% paraformaldehyde (10 min), permeabilized with 0.1% PBS-Tween for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1%PBS-Tween for 1h. The cells were then incubated overnight at 4°C with ab27053 at 5µg/ml and ab7291, Mouse monoclonal [DM1A] to alpha Tubulin - Loading Control. Cells were then incubated with ab150081, Goat polyclonal Secondary Antibody to Rabbit IgG - H&L (Alexa Fluor® 488), pre-adsorbed at 1/1000 dilution (shown in green) and ab150120, Goat polyclonal Secondary Antibody to Mouse IgG - H&L (Alexa Fluor® 594), pre-adsorbed at 1/1000 dilution (shown in pseudocolour red). Nuclear DNA was labelled with DAPI (shown in blue).
Image was acquired with a confocal microscope (Leica-Microsystems TCS SP8) and a single confocal section is shown.
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All lanes : Anti-PGP9.5 antibody (ab27053) at 1 µg/ml
Lane 1 : Human brain tissue lysate - total protein (ab29466)
Lane 2 : Brain (Mouse) Tissue Lysate (ab27253)
Lane 3 : Brain (Rat) Tissue Lysate (ab7942)
Lysates/proteins at 10 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/10000 dilution
Developed using the ECL technique.
Performed under reducing conditions.
Predicted band size: 25 kDa
Observed band size: 25 kDa
Additional bands at: 37 kDa (possible non-specific binding), 60 kDa (possible non-specific binding), 75 kDa (possible non-specific binding)
Exposure time: 10 secondsThis blot was produced using a 4-12% Bis-tris gel under the MES buffer system. The gel was run at 200V for 35 minutes before being transferred onto a Nitrocellulose membrane at 30V for 70 minutes. The membrane was then blocked for an hour using 3% Milk before being incubated with abX overnight at 4°C. Antibody binding was detected using an anti-rabbit antibody conjugated to HRP, and visualised using ECL development solution.
Abcam recommends using milk as the blocking agent.
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All lanes : Anti-PGP9.5 antibody (ab27053) at 1 µg/ml
Lane 1 : Human brain tissue lysate - total protein (ab29466)
Lane 2 : HEK293 (Human embryonic kidney cell line) Whole Cell Lysate
Lane 3 : Brain (Mouse) Tissue Lysate
Lysates/proteins at 10 µg per lane.
Secondary
All lanes : Goat polyclonal to Rabbit IgG - H&L - Pre-Adsorbed (HRP) at 1/3000 dilution
Developed using the ECL technique.
Performed under reducing conditions.
Predicted band size: 25 kDa
Observed band size: 25 kDa
Additional bands at: 65 kDa. We are unsure as to the identity of these extra bands.
Exposure time: 1 minute -
PGP9.5 was immunoprecipitated using 0.5mg Mouse Brain tissue lysate, 5µg of Rabbit polyclonal to PGP9.5 and 50µl of protein G magnetic beads (+). No antibody was added to the control (-).
The antibody was incubated under agitation with Protein G beads for 10min, Mouse Brain tissue lysate lysate diluted in RIPA buffer was added to each sample and incubated for a further 10min under agitation.
Proteins were eluted by addition of 40µl SDS loading buffer and incubated for 10min at 70oC; 10µl of each sample was separated on a SDS PAGE gel, transferred to a nitrocellulose membrane, blocked with 5% BSA and probed with ab27053.
Secondary: Clean-Blot IP Detection Reagent (HRP) at 1/500 dilution.
Band: 26kDa, non specific band - 65kDa: We are unsure as to the identity of this extra band; PGP9.5 -
Anti-PGP9.5 antibody (ab27053) at 1/250 dilution + Recombinant Human PGP9.5 protein (ab82628) at 0.01 µg
Secondary
Goat polyclonal to Rabbit IgG - H&L - Pre-Adsorbed (HRP) (ab65484) at 1/5000 dilution
Developed using the ECL technique.
Performed under reducing conditions.
Predicted band size: 25 kDa
Exposure time: 3 minutes
Protocols
Datasheets and documents
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SDS download
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Datasheet download
References (10)
ab27053 has been referenced in 10 publications.
- Han H et al. Vascular Endothelial Growth Factor Mediates the Sprouted Axonogenesis of Breast Cancer in Rat. Am J Pathol 191:515-526 (2021). PubMed: 33345997
- Ichimasu N et al. Possible involvement of type 2 cytokines in alloknesis in mouse models of menopause and dry skin. Exp Dermatol 30:1745-1753 (2021). PubMed: 34181782
- Lebonvallet N et al. A re-innervated in vitro skin model of non-histaminergic itch and skin neurogenic inflammation: PAR2-, TRPV1- and TRPA1-agonist induced functionality. Skin Health Dis 1:e66 (2021). PubMed: 35663777
- Kooij R et al. Small-Molecule Activity-Based Probe for Monitoring Ubiquitin C-Terminal Hydrolase L1 (UCHL1) Activity in Live Cells and Zebrafish Embryos. J Am Chem Soc 142:16825-16841 (2020). PubMed: 32886496
- Ramírez-García PD et al. A pH-responsive nanoparticle targets the neurokinin 1 receptor in endosomes to prevent chronic pain. Nat Nanotechnol 14:1150-1159 (2019). PubMed: 31686009
- Martins DE et al. Molecular alterations of human lumbar yellow ligament related to the process of intervertebral disk degeneration and stenosis. Eur Spine J 28:1413-1422 (2019). PubMed: 31069526
- Miller RE et al. Visualization of Peripheral Neuron Sensitization in a Surgical Mouse Model of Osteoarthritis by In Vivo Calcium Imaging. Arthritis Rheumatol 70:88-97 (2018). PubMed: 28992367
- Belkind-Gerson J et al. Colitis promotes neuronal differentiation of Sox2+ and PLP1+ enteric cells. Sci Rep 7:2525 (2017). PubMed: 28566702
- Humes DJ et al. Visceral hypersensitivity in symptomatic diverticular disease and the role of neuropeptides and low grade inflammation. Neurogastroenterol Motil 24:318-e163 (2012). PubMed: 22276853
- Yu Y et al. Evaluation of blastomere biopsy using mouse model indicates the potential high-risk of neurodegenerative disorders in the offspring. Mol Cell Proteomics : (2009). WB . PubMed: 19279043