Overview

  • Product nameAnti-PGP9.5 antibodySee all PGP9.5 primary antibodies ...
  • Description
    Rabbit polyclonal to PGP9.5
  • Tested applicationsIHC-P, IP, WB, IHC-Fr, ICC/IFmore details
  • Species reactivity
    Reacts with: Mouse, Rat, Human
  • Immunogen

    Synthetic peptide conjugated to KLH derived from within residues 150 to the C-terminus of Human PGP9.5.

    (Peptide available as ab27848.)

  • Positive control
    • This antibody gave a positive signal in the following lysates: Mouse Brain Tissue, Mouse Brain Tissue (0 days old), Rat Brain Tissue, Human Brain Tissue, HEK293 Whole Cell

Properties

Applications

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
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).Can be blocked with Human PGP9.5 peptide (ab27848).

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.

IHC-Fr Use at an assay dependent concentration.
ICC/IF Use a concentration of 5 µg/ml.

Target

  • 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 diseaseDefects in UCHL1 are the cause of Parkinson disease type 5 (PARK5) [MIM:613643]; also known as Parkinson disease autosomal dominant 5. PARK5 is a complex neurodegenerative disorder with manifestations ranging from typical Parkinson disease to dementia with Lewy bodies. Clinical features include parkinsonian symptoms (resting tremor, rigidity, postural instability and bradykinesia), dementia, diffuse Lewy body pathology, autonomic dysfunction, hallucinations and paranoia.
  • Sequence similaritiesBelongs to the peptidase C12 family.
  • Post-translational
    modifications
    O-glycosylated.
  • Cellular localizationCytoplasm. Endoplasmic reticulum membrane. About 30% of total UCHL1 is associated with membranes in brain.
  • Information by UniProt
  • Database links
  • Alternative names
    • Gracile axonal dystrophy antibody
    • Gracile axonal dystrophy antibody
    • Neuron cytoplasmic protein 9.5 antibody
    • Neuron cytoplasmic protein 9.5 antibody
    • OTTHUMP00000218137 antibody
    • OTTHUMP00000218139 antibody
    • OTTHUMP00000218140 antibody
    • OTTHUMP00000218141 antibody
    • Park 5 antibody
    • Park5 antibody
    • Park5 antibody
    • Parkinson Disease 5 antibody
    • Parkinson Disease 5 antibody
    • PGP 9.5 antibody
    • PGP 9.5 antibody
    • PGP9.5 antibody
    • PGP9.5 antibody
    • PGP95 antibody
    • Protein gene product 9.5 antibody
    • Protein gene product 9.5 antibody
    • Ubiquitin C terminal esterase L1 antibody
    • Ubiquitin C terminal esterase L1 antibody
    • Ubiquitin C terminal hydrolase (neuron specific) antibody
    • Ubiquitin C terminal hydrolase (neuron specific) antibody
    • Ubiquitin C terminal hydrolase antibody
    • Ubiquitin carboxyl terminal esterase L1 antibody
    • Ubiquitin carboxyl terminal esterase L1 antibody
    • Ubiquitin carboxyl terminal hydrolase isozyme L1 antibody
    • Ubiquitin carboxyl terminal hydrolase isozyme L1 antibody
    • Ubiquitin carboxyl-terminal hydrolase isozyme L1 antibody
    • Ubiquitin thioesterase L1 antibody
    • Ubiquitin thiolesterase antibody
    • Ubiquitin thiolesterase antibody
    • Ubiquitin thiolesterase L1 antibody
    • Ubiquitin thiolesterase L1 antibody
    • UCH L1 antibody
    • UCH L1 antibody
    • UCH-L1 antibody
    • UCHL1 antibody
    • UCHL1 antibody
    • UCHL1_HUMAN antibody
    see all

Anti-PGP9.5 antibody images

  • All lanes : Anti-PGP9.5 antibody (ab27053) at 1 µg/ml

    Lane 1 : Brain (Human) Tissue Lysate - adult normal tissue (ab29466)
    Lane 2 : Brain (Mouse) Tissue Lysate (ab27253)
    Lane 3 : Brain (Rat) Tissue Lysate (ab7942)

    Lysates/proteins at 10 µg per lane.

    Secondary
    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 seconds

    This 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.

  • IHC image of PGP9.5 staining in human pancreas formalin fixed paraffin embedded tissue section, performed on a Leica BondTM system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab27053, 5µg/ml, for 15 mins 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.

  • All lanes : Anti-PGP9.5 antibody (ab27053) at 1 µg/ml

    Lane 1 : Brain (Human) Tissue Lysate - adult normal tissue (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
    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
  • ICC/IF image of ab27053 stained PC12 cells. The cells were 4% PFA fixed (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 non-specific protein-protein interactions. The cells were then incubated with the antibody (ab27053, 5µg/ml) overnight at +4°C. The secondary antibody (green) was Alexa Fluor® 488 goat anti-rabbit IgG (H+L) used at a 1/1000 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.
  • Anti-PGP9.5 antibody (ab27053) at 1/250 dilution + Human PGP9.5 full length 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
  • 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

References for Anti-PGP9.5 antibody (ab27053)

This product has been referenced in:
  • 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 . Read more (PubMed: 19279043) »

See 1 Publication for this product

Product Wall

Application Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample Cat Tissue sections (Bladder)
Specification Bladder
Fixative Formaldehyde
Antigen retrieval step Heat mediated - Buffer/Enzyme Used: Citric acid
Permeabilization No
Blocking step BSA as blocking agent for 10 minute(s) · Concentration: 1% · Temperature: 21°C
Username

Mr. Carl Hobbs

Verified customer

Submitted Nov 26 2012

Application Immunocytochemistry/ Immunofluorescence
Sample Mouse Cell (Dorsal Root Ganglia (DRG) neurons)
Specification Dorsal Root Ganglia (DRG) neurons
Fixative Formaldehyde
Permeabilization Yes - tween 20
Blocking step Serum as blocking agent for 30 minute(s) · Concentration: 2% · Temperature: 21°C
Username

Abcam user community

Verified customer

Submitted Nov 05 2012

Application Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample Rat Tissue sections (Pancreas)
Specification Pancreas
Fixative Formaldehyde
Antigen retrieval step Heat mediated - Buffer/Enzyme Used: Citric acid
Permeabilization No
Blocking step BSA as blocking agent for 10 minute(s) · Concentration: 1% · Temperature: 21°C
Username

Mr. Carl Hobbs

Verified customer

Submitted Jun 14 2012

Application Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample Mouse Tissue sections (Skin)
Specification Skin
Fixative Paraformaldehyde
Antigen retrieval step Heat mediated - Buffer/Enzyme Used: Ctrate buffer pH 6.0
Permeabilization No
Blocking step Serum as blocking agent for 30 minute(s) · Concentration: 5% · Temperature: RT°C
Username

Abcam user community

Verified customer

Submitted Jan 06 2012

Thanks for your enquiry. According to Campbell et al (2003), PGP9.5 expresses in melanocytes in addition to neurons. Please read PMID: 14559978 for more detail. I hope this information helps you, and please do not hesitate to contact us if you hav...

Read More
Application Immunohistochemistry (Frozen sections)
Sample Rat Tissue sections (brain sections)
Specification brain sections
Fixative Paraformaldehyde
Permeabilization No
Username

Dr. Sophie Pezet

Verified customer

Submitted Jul 14 2008

Application Western blot
Sample Mouse Tissue lysate - whole (Spinal cord)
Loading amount 40 µg
Specification Spinal cord
Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5%
Username

Abcam user community

Verified customer

Submitted Feb 14 2007

Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"