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    pgp95-antibody-346ct251-ab86808.pdf

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Neuroscience Neurology process Neurodegenerative disease Parkinson's disease Parkin / PARK
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Validated using a knockout cell line

Anti-PGP9.5 antibody [346CT2.5.1] (ab86808)

  • Datasheet
  • SDS
Reviews (4) Submit a question References (2)

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Western blot - Anti-PGP9.5 antibody [346CT2.5.1] (ab86808)
  • Flow Cytometry - Anti-PGP9.5 antibody [346CT2.5.1] (ab86808)
  • Western blot - Anti-PGP9.5 antibody [346CT2.5.1] (ab86808)

Key features and details

  • Mouse monoclonal [346CT2.5.1] to PGP9.5
  • Suitable for: Flow Cyt, WB
  • Knockout validated
  • Reacts with: Rat, Human
  • Isotype: IgG1

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Overview

  • Product name

    Anti-PGP9.5 antibody [346CT2.5.1]
    See all PGP9.5 primary antibodies
  • Description

    Mouse monoclonal [346CT2.5.1] to PGP9.5
  • Host species

    Mouse
  • Tested Applications & Species

    Application Species
    Flow Cyt
    Human
    WB
    Rat
    Human
    See all applications and species data
  • Immunogen

    Recombinant full length protein corresponding to Human PGP9.5 aa 1 to the C-terminus.

  • Positive control

    • 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.
  • General notes

    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.

    If you require this antibody in a particular buffer formulation or a particular conjugate for your experiments, please contact orders@abcam.com or you can find further information here.

    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. Upon delivery aliquot and store at -20°C. Avoid freeze / thaw cycles.
  • Storage buffer

    pH: 7.40
    Preservative: 0.02% Sodium azide
    Constituent: PBS
  • Concentration information loading...
  • Purity

    Protein G purified
  • Clonality

    Monoclonal
  • Clone number

    346CT2.5.1
  • Myeloma

    Sp2/0
  • Isotype

    IgG1
  • 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

  • Alternative Versions

    • Alexa Fluor® 647 Anti-PGP9.5 antibody [346CT2.5.1] (ab199217)
    • Alexa Fluor® 488 Anti-PGP9.5 antibody [346CT2.5.1] (ab201838)
  • Compatible Secondaries

    • Goat Anti-Mouse IgG H&L (Alexa Fluor® 488) (ab150113)
    • Goat Anti-Mouse IgG H&L (HRP) (ab205719)
    • Goat anti-Mouse IgG H&L (IRDye® 800CW) preadsorbed (ab216772)
    • Goat Anti-Rabbit IgG H&L (IRDye® 680RD) preadsorbed (ab216777)
  • Isotype control

    • Mouse IgG1, kappa monoclonal [15-6E10A7] - Isotype Control (ab170190)
    • Mouse IgG1, Kappa Monoclonal [B11/6] - Isotype Control (ab91353)
  • Recombinant Protein

    • Recombinant human PGP9.5 protein (ab53962)

Applications

The Abpromise guarantee

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.

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
Flow Cyt
Human
WB
Rat
Human
All applications
Mouse
Goat
Horse
Cat
Dog
Pig
Application Abreviews Notes
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.

Notes
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.

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: 100033838 Horse
    • Entrez Gene: 7345 Human
    • Entrez Gene: 22223 Mouse
    • Entrez Gene: 396637 Pig
    • Entrez Gene: 29545 Rat
    • Omim: 191342 Human
    • SwissProt: Q9GM50 Horse
    • SwissProt: P09936 Human
    • SwissProt: Q9R0P9 Mouse
    • SwissProt: Q6SEG5 Pig
    • 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 [346CT2.5.1] (ab86808)
    Western blot - Anti-PGP9.5 antibody [346CT2.5.1] (ab86808)

    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.

  • Flow Cytometry - Anti-PGP9.5 antibody [346CT2.5.1] (ab86808)
    Flow Cytometry - Anti-PGP9.5 antibody [346CT2.5.1] (ab86808)
    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.
  • Western blot - Anti-PGP9.5 antibody [346CT2.5.1] (ab86808)
    Western blot - Anti-PGP9.5 antibody [346CT2.5.1] (ab86808)
    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.

    Secondary
    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

Protocols

  • Flow cytometry protocols
  • Western blot protocols

Click here to view the general protocols

Datasheets and documents

  • SDS download

  • Datasheet download

    Download

References (2)

Publishing research using ab86808? Please let us know so that we can cite the reference in this datasheet.

ab86808 has been referenced in 2 publications.

  • 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). 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). PubMed: 25911511

Customer reviews and Q&As

Show All Reviews Q&A
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1-4 of 4 Abreviews or Q&A

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) abreview for Anti-PGP9.5 antibody [346CT2.5.1]

Excellent
Abreviews
Abreviews
abreview image
Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Goat Tissue sections (Small intestine)
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: Citric acid
Permeabilization
No
Specification
Small intestine
Blocking step
BSA as blocking agent for 10 minute(s) · Concentration: 2% · Temperature: 21°C
Fixative
Formaldehyde
Read More

Mr. Carl Hobbs

Verified customer

Submitted Oct 14 2016

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) abreview for Anti-PGP9.5 antibody [346CT2.5.1]

Good
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 29 2012

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) abreview for Anti-PGP9.5 antibody [346CT2.5.1]

Excellent
Abreviews
Abreviews
abreview image
Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Dog Tissue sections (Small intestine)
Specification
Small intestine
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 15 2012

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) abreview for Anti-PGP9.5 antibody [346CT2.5.1]

Excellent
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 13 2012

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