Overview

  • Product name
    Anti-PGP9.5 antibody - Neuronal Marker
    See all PGP9.5 primary antibodies
  • Description
    Rabbit polyclonal to PGP9.5 - Neuronal Marker
  • Host species
    Rabbit
  • Tested applications
    Suitable for: WB, ICC/IF, IHC-Pmore details
  • Species reactivity
    Reacts with: Human
  • Immunogen

    Recombinant full length protein corresponding to Human PGP9.5.
    Database link: P09936

  • Positive control
    • Pancreas and small intestine This antibody gave a positive result when used in the following formaldehyde fixed cell lines: DU145.

Properties

Applications

Our Abpromise guarantee covers the use of ab15503 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
WB Use at an assay dependent concentration. Predicted molecular weight: 25 kDa.
ICC/IF Use a concentration of 1 µg/ml.
IHC-P 1/200. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol. Cool at RT for 20 minutes.

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

  • ICC/IF image of ab15503 stained DU145. The cells were 4% formaldehyde 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 ab15503 at 1 ug/mL overnight at +4°C. The secondary antibody (green) was DyLight® 488 goat anti-rabbit IgG (H+L) (ab96899) used at a 1/250 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.

  • Formalin-fixed, paraffin-embedded human pancreas tissue stained for PGP9.5 using ab15503 at 1/200 dilution in immunohistochemical analysis.

  • Anti-PGP9.5 antibody - Neuronal Marker (ab15503) at 1/25 dilution + Human Brain lysate

    Predicted band size: 25 kDa
    Observed band size: 26 kDa
    why is the actual band size different from the predicted?

References

This product has been referenced in:
  • Gao F  et al. The neurotrophic tyrosine kinase receptor TrkA and its ligand NGF are increased in squamous cell carcinomas of the lung. Sci Rep 8:8135 (2018). Read more (PubMed: 29802376) »
  • Tam J  et al. Reconstitution of full-thickness skin by microcolumn grafting. J Tissue Eng Regen Med N/A:N/A (2016). IHC-P ; Human . Read more (PubMed: 27296503) »
See all 6 Publications for this product

Customer reviews and Q&As

1-8 of 8 Abreviews or Q&A

Application
Immunohistochemistry (Frozen sections)
Sample
Human Tissue sections (nasal mucosa)
Specification
nasal mucosa
Blocking step
Serum as blocking agent for 30 minute(s) · Concentration: 5% · Temperature: 24°C
Fixative
Paraformaldehyde

Mr. Duc Dung Le

Verified customer

Submitted Jun 10 2016

Application
Immunohistochemistry (PFA perfusion fixed frozen sections)
Sample
Mouse Tissue sections (Jugular nodose ganglion complex (JNC))
Antigen retrieval step
None
Permeabilization
No
Specification
Jugular nodose ganglion complex (JNC)
Blocking step
Serum as blocking agent for 45 minute(s) · Concentration: 5 · Temperature: RT°C
Fixative
Zamboni

Mr. Duc Dung Le

Verified customer

Submitted Apr 14 2015

Application
Immunocytochemistry
Blocking step
1% BSA/ 10% Goat / 0.3M Glycine/ 0,1%Tween/PBS as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 1% · Temperature: 25°C
Sample
Human Cultured Cells (A431 cells)
Specification
A431 cells
Permeabilization
Yes - 0.1%Tween/PBS
Fixative
Paraformaldehyde

Abcam user community

Verified customer

Submitted Aug 29 2014

Answer

Thank you for contacting us.

According to UniGene, the RNA for this protein can be found quite abundantly in human pancreas. Therefore, it shouldn't be a problem to find a suitable antibody which has been tested in human and IHC-P.

We have two polyclonal antibodies, which have been used in various references and Abreviews. However, only

https://www.abcam.com/index.html?datasheet=15503 (or use the following: https://www.abcam.com/index.html?datasheet=15503).

has been tested by us so far in-house in pancreas tissue.

The other antibody whith a lot of refferences and Abreviews would be

https://www.abcam.com/index.html?datasheet=10404 (or use the following: https://www.abcam.com/index.html?datasheet=10404).

If they would prefer a monoclonal antibody, I would like to suggest

https://www.abcam.com/index.html?datasheet=8189 (or use the following: https://www.abcam.com/index.html?datasheet=8189).

I hope this information is helpful to you. Please do not hesitate to contact us if you need any more advice or information.

Read More
Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Western blot
Sample
Human Cell lysate - whole cell (fibroblasts)
Loading amount
30 µg
Specification
fibroblasts
Treatment
see below (Additional notes)
Gel Running Conditions
Reduced Non-Denaturing (Native) (5-16% gels)
Blocking step
BSA as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C

Abcam user community

Verified customer

Submitted Jul 11 2012

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Human Tissue sections (Skin)
Specification
Skin
Fixative
Paraformaldehyde
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: citrate buffer pH 6,0
Permeabilization
No
Blocking step
Serum as blocking agent for 30 minute(s) · Concentration: 4% · Temperature: 25°C

Abcam user community

Verified customer

Submitted Dec 08 2011

Question
Answer

Thank you for your technical questionnaire. Once again I am sorry to hear that you have been having difficulties with this antibody in sheep tissue. Whilst this antibody is yet to be tested using ovine tissue it is a surprise to find that you have been obtaining correct staining in your positive and negative controls but aberrant staining that is specific to sarcocysts and not surrounding ovine tissue. I have read your technical questionnaire and you seem to have performed many of the optimisation steps that I would recommend. The fact that you are using a negative control confirms that it cannot be unquenched peroxidase activity. Furthermore given that you are blocking in goat serum the non-specific staining cannot be attributable to the secondary antibody binding to the sarcocyst cells. The only recommendation that I could recommend is to dilute the antibody further and perform longer incubations of the antibody. This may improve the specificity. It may be that this antibody demonstrated some reactivity against an ovine protein ONLY expressed in sarcocysts and therefore is showing an interesting staining pattern as a consequence. I am sorry that I cannot be of more assistance.

Read More

Answer

Thank you for your enquiry and for completing our technical questionnaire so comprehensively. I am sorry to hear that you have been having difficulties with this antibody. We recommend an antibody dilution of 1/200 and antigen retrieval by boiling tissue sections in 10mM citrate buffer, pH 6.0 for 10 min followed by cooling at RT for 20 min. Your questionnaire details that you have been following a protocol in line with this. The expression data in the EXPASY database suggests that PGP9.5 is expressed in neurons and cells of the diffuse neuroendocrine system and their tumors in human cells. This area of research is outside my area of expertise. Please can you tell me whether you expect this protein to be expressed in ovine tracheal cells. Have you verified that the transcript is present it your cells? So far this antibody has only been tested using human cells and it may be that the epitope that the antibody recognizes is not present in the ovine homolog. I hope this information helps, please do not hesitate to contact us if you need any more advice or information.

Read More

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

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