Overview

  • Product name

    Anti-PGP9.5 antibody [EPR4118]
    See all PGP9.5 primary antibodies
  • Description

    Rabbit monoclonal [EPR4118] to PGP9.5
  • Host species

    Rabbit
  • Tested applications

    Suitable for: ICC/IF, IHC-Fr, WB, IP, IHC-P, Flow Cytmore details
  • Species reactivity

    Reacts with: Mouse, Rat, Human
  • Immunogen

    Synthetic peptide corresponding to residues in Human PGP9.5 (UniProt P09936).

  • Positive control

    • WB: Fetal brain, Y79, U87-MG, SH-SY5Y, HAP1 and 293T cell lysates; IHC-P: Human glioma, colon, and hepatocellular carcinoma tissue, Mouse colon and cerebral cortex tissue, Rat Jejunum and cerebral cortex tissue; ICC/IF: Neuro-2a cells; IP: Human fetal brain lysate; FC: SH-SY5Y and Y79 cells; IHC-Fr: Mouse cerebrum tissue.
  • General notes

     

     

    Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMab® patents.

    We are constantly working hard to ensure we provide our customers with best in class antibodies. As a result of this work we are pleased to now offer this antibody in purified format. We are in the process of updating our datasheets. The purified format is designated 'PUR' on our product labels. If you have any questions regarding this update, please contact our Scientific Support team.

    This product is a recombinant rabbit monoclonal antibody.

Properties

Applications

Our Abpromise guarantee covers the use of ab108986 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
ICC/IF 1/500.
IHC-Fr 1/250.

Heat mediated antigen retrieval using sodium citrate buffer (10mM citrate pH 6.0 + 0.05% Tween-20)

WB 1/1000 - 1/10000. Detects a band of approximately 25 kDa (predicted molecular weight: 24 kDa).
IP 1/10 - 1/100.
IHC-P 1/250 - 1/1000. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Flow Cyt 1/100 - 1/10000.

ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.

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.
  • Information by UniProt
  • Database links

  • 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

  • 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)
    Lane 4: HEK293 whole cell lysate (20 µg)

    Lanes 1 - 4: Merged signal (red and green). Green - ab108986 observed at 24 kDa. Red - loading control, ab8245, observed at 37 kDa.

    Ab108986 was shown to specifically react with UCHL1 (KO) in wild-type cells as signal was lost in UCHL1 (KO) knockout HAP1 cells. Wild-type and UCHL1 (KO) knockout samples were subjected to SDS-PAGE.  Ab108986 and ab8245 (Mouse anti GAPDH loading control) were incubated overnight at 4°C at 1000 dilution 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 (Frozen sections) analysis of mouse cerebrum tissue sections labeling PGP9.5 with Purified ab108986 at 1/250 (0.5 µg/ml).Heat mediated antigen retrieval using sodium citrate buffer (10mM citrate pH 6.0 + 0.05% Tween-20). Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) was used as the secondary antibody. Negative control: PBS instead of the primary antibody. DAPI was used as a counterstain.
  • Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized Neuro-2a (Mouse neuroblastoma cell line) cells labeling PGP9.5 with ab108986 at 1/500 dilution, followed by Goat Anti-Rabbit IgG (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution (green). Confocal image showing cytoplasmic staining on Neuro-2a cell line. The nuclear counter stain is DAPI (blue).

    The negative control is PBS only.

  • Immunohistochemical analysis of paraffin-embedded human hepatocellular carcinoma tissue labeling PGP9.5 with ab108986, followed by a ready to use Goat Anti-Rabbit IgG H&L (HRP). Positive staining on human hepatocellular carcinoma. The section was incubated with ab229902 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Counterstained with Hematoxylin. Heat mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0).

    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is a ready to use Goat Anti-Rabbit IgG H&L (HRP).

  • Flow cytometric analysis of 4% paraformaldehyde  fixed 90% methanol permeabilized Y79 (Human retinoblastoma retinoblastoma) cells labelling PGP9.5 with ab108986 at 1/20 dilution (Red) compared with a Rabbit monoclonal IgG (ab172730)  isotype control (black) and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue). A Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) at 1/2000 dilution was used as the secondary antibody.  

  • PGP9.5 was immunoprecipitated from 0.35 mg Human fetal brain lysate  with ab108986 at 1/20 dilution (0.5μg in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab108986 1/500 dilution (0.17 μg/ml). VeriBlot for IP Detection Reagent (HRP) (ab131366) was used as the secondary antibody at 1/1000 dilution.

    Lane 1: Human fetal brain lysate 10μg

    Lane 2: ab108986 IP in Human fetal brain lysate

    Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab108986 in Human fetal brain lysate.

    Blocking and dilution buffer and concentration: 5% NFDM/TBST.

    Exposure time: 1 second 

  • All lanes : Anti-PGP9.5 antibody [EPR4118] (ab108986) at 1/5000 dilution

    Lane 1 : HEK-293 (Human embryonic kidney epithelial cell) whole cell lysates
    Lane 2 : SH-SY5Y (Human neuroblastoma epithelial cell) whole cell lysates
    Lane 3 : C6 (Rat glial tumor glial cell) whole cell lysates
    Lanes 4 & 6 : PC-12 (Rat adrenal gland pheochromocytoma) whole cell lysates
    Lane 5 : Neuro-2a (Mouse neuroblastoma neuroblast) whole cell lysates
    Lane 7 : Mouse brain lysates

    Lysates/proteins at 20 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution

    Predicted band size: 24 kDa
    Observed band size: 25 kDa
    why is the actual band size different from the predicted?


    Exposure time: 10 seconds


    Blocking/Diluting buffer and concentration: 5% NFDM/TBST

  • Immunohistochemical analysis of paraffin-embedded rat cerebral cortex tissue labeling PGP9.5 with ab108986, followed by a ready to use Goat Anti-Rabbit IgG H&L (HRP). Positive staining on rat cerebral cortex.The section was incubated with ab229902 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Counterstained with Hematoxylin. Heat mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0).

    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is a ready to use Goat Anti-Rabbit IgG H&L (HRP).

  • Immunohistochemical analysis of paraffin-embedded mouse cerebral cortex tissue labeling PGP9.5 with ab108986, followed by a ready to use Goat Anti-Rabbit IgG H&L (HRP). Positive staining on mouse cerebral cortex. The section was incubated with ab229902 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Counterstained with Hematoxylin. Heat mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0).

    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is a ready to use Goat Anti-Rabbit IgG H&L (HRP).

  • Formalin-fixed, paraffin-embedded Cat lung tissue stained for PGP9.5 using ab108986 at 1/250 dilution in immunohistochemical analysis. The secondary antibody was ImmPRESS™ HRP Universal Antibody (Anti-Mouse IgG/A). Antigen retrieval: Heat mediated - Buffer/Enzyme Used: 10 mM citrate, pH6.0.

  • Overlay histogram showing SH-SY5Y cells stained with ab108986 (red line). The cells were fixed with 4% paraformaldehyde (10 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 (ab108986, 1/10000 dilution) for 30 min at 22°C. The secondary antibody used was Alexa Fluor® 488 goat anti-rabbit IgG (H&L) (ab150077) at 1/2000 dilution for 30 min at 22°C. Isotype control antibody (black line) was rabbit IgG (monoclonal) (0.1μg/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 80% methanol (5 min)/permeabilized with 0.1% PBS-Tween for 20 min used under the same conditions.
  • Immunohistochemical analysis of rat Jejunum tissue sections labeling PGP9.5 with ab108986 at a dulution of 1/1000. Sections were fixed with Formaldehyde. A Biotin conjugated Goat Anti-Rabbit IgG at 1/300 was used as the secondary antibody. Antigen retrieval was heat mediated using citric acid. 

    All nerve components of enteric plexuses appear to be very well demonstrated, particularly the fine fibres of the lamina propria and the muscularis mucosa.

    See Abreview

  • All lanes : Anti-PGP9.5 antibody [EPR4118] (ab108986) at 1/1000 dilution

    Lane 1 : Fetal brain cell lysate
    Lane 2 : Y79 cell lysate
    Lane 3 : U87-MG cell lysate
    Lane 4 : SH-SY5Y cell lysate
    Lane 5 : 293T cell lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    All lanes : HRP labelled goat anti-rabbit IgG at 1/2000 dilution

    Predicted band size: 24 kDa
    Observed band size: 25 kDa why is the actual band size different from the predicted?

  • Fomalin-fixed, paraffin-embedded Dog skin tissue stained for PGP9.5 using ab108986 at 1/500 dilution in immunohistochemical analysis. ImmPRESS™ Anti-Rabbit IgG Polymer Detection Kit was used as the secondary antibody.

    Antigen Retrieval: Heat mediated - Buffer/Enzyme Used: 10 mM citrate, pH6.0

  • ab108986 staining PGP9.5 in human colon tissue sections by Immunohistochemistry (IHC-P - paraformaldehyde-fixed, paraffin-embedded sections). Tissue was fixed with formaldehyde and blocked with 2% BSA for 10 minutes at 21°C; antigen retrieval was by heat mediation in citric acid. Samples were incubated with the primary antibody (1/500 in TBS/BSA/azide) for 16 hours at 21°C. A Biotin-conjugated goat anti-rabbit IgG polyclonal (1/300) was used as the secondary antibody.

    See Abreview

  • Immunohistochemical analysis of mouse colon tissue sections labeling PGP9.5 with ab108986 at a dulution of 1/1500. Sections were fixed with Formaldehyde. A Biotin conjugated Goat Anti-Rabbit IgG at 1/300 was used as the secondary antibody. Antigen retrieval was heat mediated using citric acid. 

    All nerve cell/fibre components of enteric plexuses are demonstrated very well.

    See Abreview

  • Immunohistochemical staining of PGP9.5 in paraffin embedded Human glioma tissue, using ab108986 at a 1/250 dilution.

References

This product has been referenced in:

  • Rajendran PS  et al. Identification of peripheral neural circuits that regulate heart rate using optogenetic and viral vector strategies. Nat Commun 10:1944 (2019). Read more (PubMed: 31028266) »
  • Tang SC  et al. Pancreatic neuro-insular network in young mice revealed by 3D panoramic histology. Diabetologia 61:158-167 (2018). Read more (PubMed: 28864913) »
See all 13 Publications for this product

Customer reviews and Q&As

1-10 of 10 Abreviews or Q&A

Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Dog Tissue sections (skin)
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: 10 mM citrate, pH6.0
Permeabilization
No
Specification
skin
Blocking step
Serum as blocking agent for 30 minute(s) · Concentration: 2.5% · Temperature: 25°C
Fixative
Formaldehyde

Abcam user community

Verified customer

Submitted Jul 19 2019

Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Human Tissue sections (liver)
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: 10 mM citrate, pH6.0
Permeabilization
No
Specification
liver
Blocking step
Serum as blocking agent for 30 minute(s) · Concentration: 2.5% · Temperature: 25°C
Fixative
Formaldehyde

Abcam user community

Verified customer

Submitted Jul 17 2019

Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Finch Tissue sections (brain)
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: 10 mM citrate, pH6.0
Permeabilization
No
Specification
brain
Blocking step
Serum as blocking agent for 30 minute(s) · Concentration: 2.5% · Temperature: 25°C
Fixative
Formaldehyde

Abcam user community

Verified customer

Submitted Jun 24 2019

Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Horse Tissue sections (placenta)
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: 10 mM citrate, pH6.0
Permeabilization
No
Specification
placenta
Blocking step
Serum as blocking agent for 30 minute(s) · Concentration: 2.5% · Temperature: 25°C
Fixative
Formaldehyde

Abcam user community

Verified customer

Submitted May 20 2019

Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Mouse Tissue sections (foot pad)
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: 10 mM citrate, pH6.0
Permeabilization
No
Specification
foot pad
Blocking step
Serum as blocking agent for 30 minute(s) · Concentration: 2.5% · Temperature: 25°C
Fixative
Formaldehyde

Abcam user community

Verified customer

Submitted Apr 12 2019

Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Cat Tissue sections (lung)
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: 10 mM citrate, pH6.0
Permeabilization
No
Specification
lung
Blocking step
Serum as blocking agent for 30 minute(s) · Concentration: 2.5% · Temperature: 25°C
Fixative
Formaldehyde

Abcam user community

Verified customer

Submitted Apr 12 2019

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

Mr. Carl Hobbs

Verified customer

Submitted Dec 21 2016

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

Mr. Carl Hobbs

Verified customer

Submitted Dec 21 2016

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

Mr. Carl Hobbs

Verified customer

Submitted Dec 21 2016

Answer

Thank you for your inquiry. Unfortunately, we do not have any conjugated anti-PGP9.5 antibodies available in our catalog. Having said this, I am pleased to let you know that we have conjugation kits available that allow a quick and easy process to conjugate an antibody of your choice, in whatever function you need. It has the advantage of being a one-step antibody labeling method with no separation steps. Please find more detailed information under https://www.abcam.com/EasyLink or in the attached guide. I hope this information is nevertheless helpful to you. Please do not hesitate to contact me if you have any further questions in this regard.

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