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Validated using a knockout cell line

Anti-PGP9.5 antibody (ab27053)

  • Datasheet
  • SDS
Reviews (6)Q&A (1)References (5)

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Western blot - Anti-PGP9.5 antibody (ab27053)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PGP9.5 antibody (ab27053)
  • Western blot - Anti-PGP9.5 antibody (ab27053)
  • Immunocytochemistry/ Immunofluorescence - Anti-PGP9.5 antibody (ab27053)
  • Western blot - Anti-PGP9.5 antibody (ab27053)
  • Immunoprecipitation - Anti-PGP9.5 antibody (ab27053)
  • Western blot - 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

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Overview

  • Product name

    Anti-PGP9.5 antibody
    See all PGP9.5 primary antibodies
  • Description

    Rabbit polyclonal to PGP9.5
  • Host species

    Rabbit
  • Tested Applications & Species

    Application Species
    ICC/IF
    Rat
    IHC-P
    Human
    IP
    Mouse
    WB
    Mouse
    Rat
    Human
    See all applications and species data
  • Immunogen

    Synthetic peptide corresponding to Human PGP9.5 aa 150 to the C-terminus conjugated to keyhole limpet haemocyanin.

  • 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 the problem with our range of recombinant monoclonal antibodies and knockout edited cell lines for gold-standard validation.

    One factor contributing to the crisis is the use of antibodies that are not suitable. This can lead to misleading results and the use of incorrect data informing project assumptions and direction. To help address this challenge, we have introduced an application and species grid on our primary antibody datasheets to make it easy to simplify identification of the right antibody for your needs.

    Learn more here.

Properties

  • 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...
  • Purity

    Immunogen affinity purified
  • Clonality

    Polyclonal
  • Isotype

    IgG
  • Research areas

    • Neuroscience
    • Neurology process
    • Neurodegenerative disease
    • Parkinson's disease
    • Parkin / PARK
    • Neuroscience
    • Cell Type Marker
    • Neuron marker
    • Soma marker
    • Cell Biology
    • Proteolysis / Ubiquitin
    • Proteasome / Ubiquitin
    • Deubiquitination
    • Tags & Cell Markers
    • Cell Type Markers
    • Neuroscience Markers
    • Neuronal

Associated products

  • Compatible Secondaries

    • Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077)
    • Goat Anti-Rabbit IgG H&L (HRP) (ab205718)
    • Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773)
    • Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (ab216776)
  • Isotype control

    • Rabbit IgG, polyclonal - Isotype Control (ChIP Grade) (ab171870)
  • Positive Controls

    • Recombinant human PGP9.5 protein (ab53962)
  • Recombinant Protein

    • Recombinant human PGP9.5 protein (ab53962)

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.

Guaranteed

Tested applications are guaranteed to work and covered by our Abpromise guarantee.

Predicted

Predicted to work for this combination of applications and species but not guaranteed.

Incompatible

Does not work for this combination of applications and species.

Application Species
ICC/IF
Rat
IHC-P
Human
IP
Mouse
WB
Mouse
Rat
Human
Application Abreviews Notes
IHC-P (3)
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 (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 5 µg/ml.
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).

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 5 µg/ml.

Target

  • 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
    modifications

    O-glycosylated.
  • Cellular localization

    Cytoplasm. Endoplasmic reticulum membrane. About 30% of total UCHL1 is associated with membranes in brain.
  • Target information above from: UniProt accession P09936 The UniProt Consortium
    The Universal Protein Resource (UniProt) in 2010
    Nucleic Acids Res. 38:D142-D148 (2010) .

    Information by UniProt
  • 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
    • Unigene: 29807 Mouse
    • Unigene: 107213 Rat
    see all
  • Alternative names

    • Epididymis luminal protein 117 antibody
    • Epididymis secretory protein Li 53 antibody
    • HEL 117 antibody
    • HEL S 53 antibody
    • NDGOA antibody
    • Neuron cytoplasmic protein 9.5 antibody
    • OTTHUMP00000218137 antibody
    • OTTHUMP00000218139 antibody
    • OTTHUMP00000218140 antibody
    • OTTHUMP00000218141 antibody
    • Park 5 antibody
    • PARK5 antibody
    • PGP 9.5 antibody
    • PGP9.5 antibody
    • PGP95 antibody
    • Protein gene product 9.5 antibody
    • Ubiquitin C terminal esterase L1 antibody
    • Ubiquitin C terminal hydrolase antibody
    • Ubiquitin C terminal hydrolase L1 antibody
    • Ubiquitin carboxyl terminal esterase 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 L1 antibody
    • UCH-L1 antibody
    • UCHL1 antibody
    • UCHL1_HUMAN antibody
    see all

Images

  • Western blot - Anti-PGP9.5 antibody (ab27053)
    Western blot - Anti-PGP9.5 antibody (ab27053)

    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.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PGP9.5 antibody (ab27053)
    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.

  • Western blot - Anti-PGP9.5 antibody (ab27053)
    Western blot - Anti-PGP9.5 antibody (ab27053)
    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 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.

  • Immunocytochemistry/ Immunofluorescence - Anti-PGP9.5 antibody (ab27053)
    Immunocytochemistry/ Immunofluorescence - Anti-PGP9.5 antibody (ab27053)
    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.
  • Western blot - Anti-PGP9.5 antibody (ab27053)
    Western blot - Anti-PGP9.5 antibody (ab27053)
    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
  • Immunoprecipitation - Anti-PGP9.5 antibody (ab27053)
    Immunoprecipitation - Anti-PGP9.5 antibody (ab27053)
    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
  • Western blot - Anti-PGP9.5 antibody (ab27053)
    Western blot - Anti-PGP9.5 antibody (ab27053)
    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

  • Immunoprecipitation protocols
  • Immunohistochemistry protocols
  • Immunocytochemistry & immunofluorescence protocols
  • Western blot protocols

Click here to view the general protocols

Datasheets and documents

  • SDS download

  • Datasheet download

    Download

References (5)

Publishing research using ab27053? Please let us know so that we can cite the reference in this datasheet.

ab27053 has been referenced in 5 publications.

  • 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
  • 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
  • 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

Customer reviews and Q&As

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1-7 of 7 Abreviews or Q&A

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) abreview for Anti-PGP9.5 antibody - Neuronal Marker

Average
Abreviews
Abreviews
abreview image
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
Read More

Mr. Carl Hobbs

Verified customer

Submitted Nov 26 2012

Immunocytochemistry/ Immunofluorescence abreview for Anti-PGP9.5 antibody - Neuronal Marker

Good
Abreviews
Abreviews
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
Read More

Abcam user community

Verified customer

Submitted Nov 05 2012

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) abreview for Anti-PGP9.5 antibody - Neuronal Marker

Average
Abreviews
Abreviews
abreview image
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
Read More

Mr. Carl Hobbs

Verified customer

Submitted Jun 14 2012

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) abreview for Anti-PGP9.5 antibody - Neuronal Marker

Good
Abreviews
Abreviews
abreview image
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
Read More

Abcam user community

Verified customer

Submitted Jan 06 2012

Question

I stained a paraffin embedded skin tissue section with ab27053, PGP9.5 antibody - Neuronal Marker. It seems like not only sensory neurons but also melanocytes got staining with ab27053. Is it an appropriate staining figure for this antibody?

Read More

Abcam community

Verified customer

Asked on Jan 14 2010

Answer

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 have further questions.

Read More

Abcam Scientific Support

Answered on Jan 14 2010

Immunohistochemistry (Frozen sections) abreview for Anti-PGP9.5 antibody - Neuronal Marker

Excellent
Abreviews
Abreviews
abreview image
Application
Immunohistochemistry (Frozen sections)
Sample
Rat Tissue sections (brain sections)
Specification
brain sections
Fixative
Paraformaldehyde
Permeabilization
No
Read More

Dr. Sophie Pezet

Verified customer

Submitted Jul 14 2008

Western blot abreview for Anti-PGP9.5 antibody - Neuronal Marker

Good
Abreviews
Abreviews
abreview image
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%
Read More

Abcam user community

Verified customer

Submitted Feb 14 2007

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