Overview

  • Product name
  • Description
    Rabbit polyclonal to PGP9.5
  • Host species
    Rabbit
  • Tested applications
    Suitable for: IHC-Fr, WB, IHC-P, IHC (PFA fixed), ICC, ICC/IFmore details
  • Species reactivity
    Reacts with: Mouse, Rat, Human, Pig
    Predicted to work with: Horse
  • Immunogen

    Synthetic peptide:

    ASSEDTLLKDAAKVCR

    , corresponding to amino acids 176 - 191 of Human PGP9.5.

  • Positive control
    • Recombinant human PGP9.5 protein (ab53962) can be used as a positive control in WB. Human brain lysate.
  • General notes
    Serum may become trapped in top of vial during shipping. Centrifugation of vial is recommended before opening.

Properties

Applications

Our Abpromise guarantee covers the use of ab10404 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-Fr 1/2000.
WB 1/1000. Predicted molecular weight: 26.8 kDa.Can be blocked with Human PGP9.5 peptide (ab38203).
IHC-P 1/100 - 1/300.
IHC (PFA fixed) 1/200. PubMed: 17508253
ICC Use at an assay dependent concentration.
ICC/IF 1/1000.

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

  • ab10404 at a 1/500 dilution staining human cancer prostate tissue sections by Immunohistochemistry (Formalin-fixed paraffin-embedded sections).Following heat mediated antigen retrieval the tissue was fixed with formaldehyde and blocked with 20% serum. The tissue was incubated with the primary antibody for 1 hour and then with an HRP conjugated rabbit polyclonal antibody.

    See Abreview

  • Immunofluorescence analysis of mouse gonocytes, staining PGP9.5 with ab10404.

    Cells were permabilized with 0.025% Triton X-100 and blocked with 1% BSA for 1 hour at room temperature. Cells were incubated with primary antibody (1/20) overnight at 4°C. An AlexaFluor®594-conjugated anti-rabbit IgG was used as the secondary antibody.
  • Anti-PGP9.5 antibody (ab10404) at 1/1000 dilution + Human brain tissue lysate - total protein (ab29466) at 10 µg

    Secondary
    Goat polyclonal to Rabbit IgG - H&L - Pre-Adsorbed (HRP) at 1/3000 dilution

    Predicted band size: 26.8 kDa
    Observed band size: 27 kDa
    why is the actual band size different from the predicted?
    Additional bands at: 48 kDa, 70 kDa, 73 kDa. We are unsure as to the identity of these extra bands.

  • ab10404 at a dilution of 1/1000, detecting PGP9.5 in cortical neurons (Alexa 488 secondary at 1/2000) on rat brain tissue sections prepared for free floating IHC (4% PFA fixed, 30µm thick coronal sections; see protocol link for detailed description). Image taken with a 20x objective. No labeling observed following omission of primary antibody.

    Sections were viewed using an Axioplan 2 Imaging microscope (Imaging Associates) fitted with 10x, 20x and 40x Plan-Neofluorobjectives (Zeiss, Germany) and images were taken using a AxioCam Hrm digital camera (Zeiss, Germany) and AxioVision software (Imaging Associates).

  • ab10404 staining PGP9.5 in Mouse Endometriotic tissue sections by Immunohistochemistry (IHC-P - paraformaldehyde-fixed, paraffin-embedded sections). Tissue was fixed with paraformaldehyde and blocked with 3% serum for 1 hour at 25°C; antigen retrieval was by heat mediation in a citrate buffer. Samples were incubated with primary antibody (1/200 in 3% BSA) for 20 hours at 4°C. An undiluted Biotin-conjugated Goat anti-rabbit IgG polyclonal was used as the secondary antibody.

    See Abreview

  • ab10404 at a dilution of 1/2000, staining PGP9.5 in rat dorsal root ganglion by Immunohistochemistry (PFA perfusion fixed frozen sections).

References

This product has been referenced in:
  • Dias DO  et al. Reducing Pericyte-Derived Scarring Promotes Recovery after Spinal Cord Injury. Cell 173:153-165.e22 (2018). Read more (PubMed: 29502968) »
  • Chan KY  et al. Engineered AAVs for efficient noninvasive gene delivery to the central and peripheral nervous systems. Nat Neurosci 20:1172-1179 (2017). IHC ; Mouse . Read more (PubMed: 28671695) »
See all 22 Publications for this product

Customer reviews and Q&As

1-10 of 15 Abreviews or Q&A

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Rat 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

Abcam guarantees this product to work in the species/application used in this Abreview.
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

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 (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

Question
Answer

Thank you for your question. The protein PGP9.5 is expressed in neurons and most brain tissue, especially from the cerebral cortex, will be a good positive tissue control. A negative tissue control is more difficult to recommend but the  Human Protein Atlas has RNA and protein expression data for tissues at http://www.proteinatlas.org/ENSG00000154277-UCHL1/tissue, and cell line data at http://www.proteinatlas.org/ENSG00000154277-UCHL1/cell/HPA005993.

Read More

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
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Human Tissue sections (Endometriotic tissue)
Specification
Endometriotic tissue
Fixative
Paraformaldehyde
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: Citrate buffer
Permeabilization
No
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 3% · Temperature: 25°C

Dr. Brett Mckinnon

Verified customer

Submitted Jan 15 2013

Answer

Thank you for contacting us.

ab10404 is a whole serum product and the specific antibody concentration cannot be determined.

I am sorry for not being able to provide useful information this time. Please do not hesitate to contact us if you need any more advice or information in other aspects.

Use our products? Submit an Abreview. Earn rewards!
https://www.abcam.com/abreviews

Read More

Question

Hi, I`ve been tried to dilute the antibody to 1:20000, but the same
result was obteined, I think that the antibody solution is
contaminated or impure. Here is the details requested. I Add a picture
in which is posible apreciate the high inespecific signal. I`ll wait
for your answer.
Best.

*Order Details
Antibody code: ab10404
Lot number: GR24090-6
Purchase order number
or preferably Abcam order number:
General Information
Antibody storage conditions (temperature/reconstitution etc): The antibody
was alicuoted and stored at -20°C
Description of the problem (high background, low signal, non-specific
satining etc.): High background and inespecific marked cells or
non-specific staining.
Sample (Species/Tissue/Cell Type/Cell Line etc.): Human spinal cord and
eutopic and ectopic endometria, both fixed in formaldehyde 10%v/v in buffer
PBS and embedded in paraffin.
Fixation of sample
(Ethanol/Methanol/Acetone/Paraformaldehyde/Other/Duration etc.)
Formaldehyde 10%v/v in buffer PBS
Antigen retrieval (Enzymatic method, Heat mediated technique etc.)
The sections were performed with antigen retrieval: Heat mediated with
vegetable steamer in pH6.0 sodium citrate buffer, and pH 8.0 EDTA
during 20, 30 and 40
minutes; in addition, the IHC was also performed without antigen retrieval
and the result was the same.
Permeabilization step: It is not necesary for tissue sections
Blocking conditions (Buffer/time period, Blocking agent etc.): It was used
2,5% normal horse serum blocking agent from the vectastain kit: Elite ABC
kit PK-7200 (Vector), and 2% bovine serum albumin for 30 and 40 minutes.
Primary Antibody (Manufacturer/Species/Diluent/Dilution/Incubation time,
Wash step): Dilutions were tested from 1:100 to 1:20000 in PBS buffer and in
1% BSA, incubating one hour at 37oC and 20 hours at 4oC. Then, the sections
was washed in PBS buffer 3 times, 3 minutes each.
Secondary Antibody (Manufacturer/Species/Diluent/Dilution/Incubation time,
Wash step): It was used the R.T.U. biotinylated universal antibody
anti-rabbit/mouse IgG, made in horse from the **Elite ABC kit PK-7200
(Vector), during 30 minutes and then washed in PBS 3 times, 3 minutes each.*
*
Detection method: ABC method, using the vectastain R.T.U. Elite ABC reagent
from **the **Elite ABC kit PK-7200 (vector)* *
Positive and negative controls used (please specify): It was used
formaldehyde fixed and parafin embedded human spinal cord sections to the
positive control, and one section from each slide stained was used to the
negative control without the primary antibody incubation and the results of
this negative controls were satisfactory in all cases.
Optimization attempts (problem solving)
How many times have you tried the IHC?
10 times
Have you run a "No Primary" control?
Yes
Do you obtain the same results every time?
Yes
What steps have you altered?
Time and temperature incubation, with (in different times) and without
antigen retrieval, different blocking solutions and in different times and
different dilutions of the antibody
Additional Notes
We would appreciate if you are also able to provide and image which woudl
help us to assess the results*

Read More
Answer

Thank you for taking the time to complete our questionnaire.

The details you have kindly provided will enable us to investigate this case for you and this is also helpful in our records for monitoring of quality.

Reviewing this case, I would like to offer some suggestions to help optimize the results from ab10404. I would also appreciate if you can confirm some further details:

1. I am sorry I am not able to trace any orders for this antibody from Chile.
Please confirm the order number and date of purchase and details of where you purchased from. If you have ordered through a distributor, they will have the order number reference from us.

2. I can recommend to try different time points for antigen retrieval as this can sometimes help optimize the results. For example, try 3, 5, 10 and 20 minutes.

3. I can recommend to continue to use a lower concentration of the antibody, 1:10000 or1:20000. However, I suggest to try adding 0.2% Tween to the antibody dilution buffers and the wash buffers. This will help to solubilise the antibody and also help to wash away any excess antibody.

4. We recommend not to mix blocking agents in one experiment as it can affect the results. I can suggestincreasing the amount of blocking agent in the blocking buffer, try 5% or 10% serum only. And alsoadd 2% Serum to the antibody dilution buffer.

5. Please confirm if the secondary antibody is working well with other rabbit primary antibodies? I can suggest it would be important to consider including a no primary control to assess if the secondary antibody is binding non specifically. What were the results from the no primary control? The concentration of the secondary may need to be reduced to help optimize the results.

I hope this information is helpful, thank you for your cooperation. Should the suggestions not improve the results, please do not hesitate to contact me again with the further requested details including the results of the no primary control.

Read More

Question
Answer

TThank you for calling Abcam regarding positive controls.

AB10404 has been used in IHC-P on lung sections in the following publication:Liu Yet al.Human metapneumovirus establishes persistent infection in the lungs of mice and is reactivated by glucocorticoid treatment.J Virol83:6837-48 (2009).http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&list_uids=19357175&dopt=Abstract.
This product has also been used by customers in human prostate carcinoma tissue to good effect. More information about this can be seen at this link:https://www.abcam.com/index.html?datasheet=10404&tab=abreviews&intabreviewid=5255

In regards to AB8874: The publication,Liu Set al.Cross-talk between Schwann cells and neuroblasts influences the biology of neuroblastoma xenografts.Am J Pathol166:891-900 (2005).http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&list_uids=15743800&dopt=Abstract,uses human schwannoma as a positive control for anti-human p75NGFRantibody.

I hope that this information is helpful. Please let me know if you have any questions.

Read More

Answer

Thank you for taking the time to contact us. I am sorry to hear you have had difficulty obtaining satisfactory results from this antibody.

I appreciate the time you have spent in the laboratory and understand your concerns. It is regrettable the results have not been successful.

I would like to reassure you that this antibody is tested and covered by our guarantee forIHC-P. I am sorry Ido not know the species you are testing but it is guaranteed for Mouse, Rat, HumanandPig. Before deciding how to proceed, I would like to investigate this particular case further for you, and also obtain some further information for our quality records. In order to do this, I have enclosed a questionnaire below. I would appreciate if you could complete this. It will help you put the information we require together very easily.

We will happily offer technical support and in the event that a product is not functioning in the applications and speciescited on the product data sheet (and the problem has been reported within 6 months of purchase), we will be pleased to provide a credit note, free of charge replacement or refund.

Thank you for your time and cooperation. We look forward to receiving the completed questionnaire.

Order Details
Antibody code:

Lot number:

Purchase order number
or preferably Abcam order number:


General Information
Antibody storage conditions (temperature/reconstitution etc)


Description of the problem (high background, low signal, non-specific satining etc.)

Sample (Species/Tissue/Cell Type/Cell Line etc.)


Fixation of sample (Ethanol/Methanol/Acetone/Paraformaldehyde/Other/Duration etc.)


Antigen retrieval (Enzymatic method, Heat mediated technique etc.)


Permeabilization step


Blocking conditions (Buffer/time period, Blocking agent etc.)


Primary Antibody (Manufacturer/Species/Diluent/Dilution/Incubation time, Wash step)


Secondary Antibody (Manufacturer/Species/Diluent/Dilution/Incubation time, Wash step)


Detection method


Positive and negative controls used (please specify)


Optimization attempts (problem solving)
How many times have you tried the IHC?



Have you run a "No Primary" control?
Yes No

Do you obtain the same results every time?
Yes No


What steps have you altered?


Additional Notes


We would appreciate if you are also able to provide and image which woudl help us to assess the results

Read More

1-10 of 15 Abreviews or Q&A

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