Overview

  • Product name
    Anti-Prion protein PrP antibody
    See all Prion protein PrP primary antibodies
  • Description
    Goat polyclonal to Prion protein PrP
  • Host species
    Goat
  • Tested applications
    Suitable for: IHC-FoFr, WB, ELISA, Dot blot, IHC-Pmore details
  • Species reactivity
    Reacts with: Mouse, Sheep, Hamster, Cow, Human
  • Immunogen

    Synthetic peptide, corresponding to amino acids 79-97 of Human Prion protein PrP.

  • Positive control
    • CJD brain.
  • General notes
    Prion protein has recently been classified as CD230 at the 7th HLDA workshop.

Properties

Applications

Our Abpromise guarantee covers the use of ab6664 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-FoFr Use at an assay dependent dilution. PubMed: 16492732
WB Use at an assay dependent dilution.
ELISA Use at an assay dependent dilution. It has only been tested for direct ELISA but not for Sandwich ELISA.
Dot blot Use at an assay dependent dilution.
IHC-P 1/200. Do not perform antigen retrieval.

Target

  • Function
    The function of PrP is still under debate. May play a role in neuronal development and synaptic plasticity. May be required for neuronal myelin sheath maintenance. May play a role in iron uptake and iron homeostasis (By similarity). Isoform 2 may act as a growth suppressor by arresting the cell cycle at the G0/G1 phase. Soluble oligomers are toxic to cultured neuroblastoma cells and induce apoptosis (in vitro).
  • Involvement in disease
    Note=PrP is found in high quantity in the brain of humans and animals infected with neurodegenerative diseases known as transmissible spongiform encephalopathies or prion diseases, like: Creutzfeldt-Jakob disease (CJD), fatal familial insomnia (FFI), Gerstmann-Straussler disease (GSD), Huntington disease-like type 1 (HDL1) and kuru in humans; scrapie in sheep and goat; bovine spongiform encephalopathy (BSE) in cattle; transmissible mink encephalopathy (TME); chronic wasting disease (CWD) of mule deer and elk; feline spongiform encephalopathy (FSE) in cats and exotic ungulate encephalopathy (EUE) in nyala and greater kudu. The prion diseases illustrate three manifestations of CNS degeneration: (1) infectious (2) sporadic and (3) dominantly inherited forms. TME, CWD, BSE, FSE, EUE are all thought to occur after consumption of prion-infected foodstuffs.
    Defects in PRNP are the cause of Creutzfeldt-Jakob disease (CJD) [MIM:123400]. CJD occurs primarily as a sporadic disorder (1 per million), while 10-15% are familial. Accidental transmission of CJD to humans appears to be iatrogenic (contaminated human growth hormone (HGH), corneal transplantation, electroencephalographic electrode implantation, etc.). Epidemiologic studies have failed to implicate the ingestion of infected annimal meat in the pathogenesis of CJD in human. The triad of microscopic features that characterize the prion diseases consists of (1) spongiform degeneration of neurons, (2) severe astrocytic gliosis that often appears to be out of proportion to the degree of nerve cell loss, and (3) amyloid plaque formation. CJD is characterized by progressive dementia and myoclonic seizures, affecting adults in mid-life. Some patients present sleep disorders, abnormalities of high cortical function, cerebellar and corticospinal disturbances. The disease ends in death after a 3-12 months illness.
    Defects in PRNP are the cause of fatal familial insomnia (FFI) [MIM:600072]. FFI is an autosomal dominant disorder and is characterized by neuronal degeneration limited to selected thalamic nuclei and progressive insomnia.
    Defects in PRNP are the cause of Gerstmann-Straussler disease (GSD) [MIM:137440]. GSD is a heterogeneous disorder and was defined as a spinocerebellar ataxia with dementia and plaquelike deposits. GSD incidence is less than 2 per 100 million live births.
    Defects in PRNP are the cause of Huntington disease-like type 1 (HDL1) [MIM:603218]. HDL1 is an autosomal dominant, early onset neurodegenerative disorder with prominent psychiatric features.
    Defects in PRNP are the cause of kuru (KURU) [MIM:245300]. Kuru is transmitted during ritualistic cannibalism, among natives of the New Guinea highlands. Patients exhibit various movement disorders like cerebellar abnormalities, rigidity of the limbs, and clonus. Emotional lability is present, and dementia is conspicuously absent. Death usually occurs from 3 to 12 month after onset.
    Defects in PRNP are the cause of spongiform encephalopathy with neuropsychiatric features (SENF) [MIM:606688]; an autosomal dominant presenile dementia with a rapidly progressive and protracted clinical course. The dementia was characterized clinically by frontotemporal features, including early personality changes. Some patients had memory loss, several showed aggressiveness, hyperorality and verbal stereotypy, others had parkinsonian symptoms.
  • Sequence similarities
    Belongs to the prion family.
  • Domain
    The normal, monomeric form has a mainly alpha-helical structure. The disease-associated, protease-resistant form forms amyloid fibrils containing a cross-beta spine, formed by a steric zipper of superposed beta-strands. Disease mutations may favor intermolecular contacts via short beta strands, and may thereby trigger oligomerization.
    Contains an N-terminal region composed of octamer repeats. At low copper concentrations, the sidechains of His residues from three or four repeats contribute to the binding of a single copper ion. Alternatively, a copper ion can be bound by interaction with the sidechain and backbone amide nitrogen of a single His residue. The observed copper binding stoichiometry suggests that two repeat regions cooperate to stabilize the binding of a single copper ion. At higher copper concentrations, each octamer can bind one copper ion by interactions with the His sidechain and Gly backbone atoms. A mixture of binding types may occur, especially in the case of octamer repeat expansion. Copper binding may stabilize the conformation of this region and may promote oligomerization.
  • Post-translational
    modifications
    The glycosylation pattern (the amount of mono-, di- and non-glycosylated forms or glycoforms) seems to differ in normal and CJD prion.
    Isoform 2 is sumoylated by SUMO1.
  • Cellular localization
    Cell membrane. Golgi apparatus and Cytoplasm. Nucleus. Accumulates outside the secretory route in the cytoplasm, from where it relocates to the nucleus.
  • Information by UniProt
  • Database links
  • Alternative names
    • Alternative prion protein; major prion protein antibody
    • AltPrP antibody
    • ASCR antibody
    • CD230 antibody
    • CD230 antigen antibody
    • CJD antibody
    • GSS antibody
    • KURU antibody
    • Major prion protein antibody
    • p27 30 antibody
    • PRIO_HUMAN antibody
    • Prion protein antibody
    • Prion related protein antibody
    • PRIP antibody
    • PRNP antibody
    • PrP antibody
    • PrP27 30 antibody
    • PrP27-30 antibody
    • PrP33-35C antibody
    • PrPC antibody
    • PrPSc antibody
    • Sinc antibody
    see all

Images

  • CJD brain section showing stained prion plaque using ab6664 at 1:200

References

This product has been referenced in:
  • Piccardo P  et al. Dissociation of prion protein amyloid seeding from transmission of a spongiform encephalopathy. J Virol 87:12349-56 (2013). Read more (PubMed: 24027305) »
  • Steele AD  et al. Prion protein (PrPc) positively regulates neural precursor proliferation during developmental and adult mammalian neurogenesis. Proc Natl Acad Sci U S A 103:3416-21 (2006). ICC/IF, IHC-FoFr ; Mouse . Read more (PubMed: 16492732) »
See all 2 Publications for this product

Customer reviews and Q&As

1-10 of 12 Abreviews or Q&A

Answer

Thank you for kindly confirming these details.

As requested, we would be pleased to arrange this free of charge replacement. In order to do this, I am forwarding these details on to our distributors team who will be happy to organize this for you.

Original order number: ######

Original order: 1 X vial ab6664
Please provide alternative FOCR for the customer: ab61409

I would like to reassure you that this free of charge replacement vial is also covered by our Abpromise guarantee. Should you still be experiencing difficulties, or if you have any further questions, please do not hesitate to let me know.

Thank you for your help and cooperation with this case. Please do not hesitate to contact us if you need anything further.

Read More

Answer

Thank you for your message and for providing this further information.

I am sorry to hear the suggestions made have not improved the results on this occasion. I appreciate the time you have spent on these experiments and would be pleased to arrange a free of charge replacement or credit note in compensation.

I look forward to hearing from you with details of how you would like to proceed.

Read More

Answer

Thank you for your reply and for kindly confirming this further information. This enquiry has been forwarded to me as my colleague is currently away from the office.

I appreciate your cooperation and understand your concerns. It is regrettable the results have not been succesful.

In order to help with our investigation of this case before deciding how to proceed, I would appreciate if you could confirm if the antbody has been tried at a lower concentration as suggested previously? For example, the customer could try 1:3000 or even 1:5000. This may help to reduce any non specific binding.

Thank you for your time. I look forward to hearing from you with the requested information and hope we can resolve this case as soon as possible.

Read More

Answer

Thank you for your reply and providing that extra information.

I am happy to send you ab73098 as a free of charge replacement as ab7303 also did not prove successful for you. Your new order number is ****** and currently we are out of stock of ab73098 but it due to come back in stock in early March and we will ship it you as soon as possible (hopefully you should have it by 7th March).

Please let me know if there is anything else I can help you with

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Answer

Thank you for your reply.

I am sorry that the replacement antibody that we sent did not prove to be successful for you. I know that you mentioned that you tried different antigen retrieval protocols, but did you also try different antibody concentrations? If so, what concentrations did you try?

Also I remember during our original correspondence raised concerns about the fixation that you were required to use on your samples could be affecting your chances of getting positive staining. I was wondering if you had gotten any other antibodies to give a positive signal in your tissue. I ask because if you would like to receive ab73098 as another replacement antibody, I want to make sure there is a very good chance of it working for you in your samples.

I look forward to your reply.

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Answer

Thank you for your reply.

I have processed your request to receive ab7303 as a replacement for ab6664 which did not work in IHC-P. Currently ab7303 is out of stock, but we have an estimated delivery date of 14th February and your new order number is *********.

Please let me know if there is anything else I can help you with.

Read More

Answer

Thank you for your reply.

I am sorry that the O/N incubation did not prove to be successful, I could be as you initially suspected and that it is the formic acid fix that is the problem.

I am happy to send you ab7303 to see if that will prove to be more successful for you, and you are correct that the immunogen sequence for this antibody is between 79-97AA of Prion protein PrP. From what I can see ab93273 & abab664 are against the same target but are from different animals and so will be slightly different.

If you would like to try out ab7303, then please let me know and I will send it to you, or if you prefer to receive a different primary antibody then that is fine too.

If you could send me the original order information when you reply, along with the adID of the product you with to receive, I will get the replacement antibody shipped to you as soon as possible.

I look forward to your reply.

Read More

Question

Find our protocol below. The only thing I could think of is the formic acid. But according to safety protocols we are not allowed to work without. We urgently need an antibody to detect Jakob-Creutzfeld-Disease. So I am looking forward if you could help us.

1) Fixation step - Formic Acid

Fixation time 60 min



Fixation temperature roomtemperature



2) Antigen retrieval method (time and procedure)-heat unmasking, heat unmasking + proteinase K treatment and protease treatment

1. without antigen retrieval – only background staining

2. with protease typ XXIV 0.1 % in PBS for 10 min – quite impressive staining of vessel wall

3. microwavetreatment: Citratebuffer pH 6.0 – granular staining in neurons (but also in normal patients)

4. microwavetreatment: Citratebuffer pH 9.0 – similar to pH 6.0

5. Proteinase K treatment (Dako S 3004) 1:500 in PBS for 10 min RT, followed by microwavetreatment Citratebuffer pH 6.0 – only background


3) Permeabilization method: No


4) Blocking agent (eg BSA, serum…): No


5) Endogenous peroxidases blocked? Dako Real Peroxidase Blocking Solution 10 min RT


Endogenous biotins blocked? No


6) Primary antibody (If more than one was used, describe in “additional notes”) :


Concentration or dilution 1:100, 1:200, 1:500


Diluent buffer Dako Real Antibody Diluent


Incubation time 60 min RT


7) Secondary antibody: Rabbit anti Goat IgG-Fc Bethyl Laboratories



Concentration or dilution 1:1000 (tested and works for other routine antibodies)



Diluent buffer Dako Real Antibody Diluent



Incubation time 30 min


8) Washing after primary and secondary antibodies:



Buffer PBS



Number of washes 3 times 5 minutes



9) Detection method Dako Real Envision with DAB as chromogen



10) How many times have you run this staining? Two times



Do you obtain the same results every time? Yes

Read More
Answer

Thank you for your reply.

Based on the information you provided and the extensive troubleshooting you have already done using this antibody, I agree that the most probable cause is the formic acid. As the most successful protocols using this antibody have been by using PFA (http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&list_uids=16492732&dopt=Abstract). However one major difference between the protocol you are using and the PubMed reference is that they did an overnight incubation at 4C instead of only 60 minutes at room temp. I would therefore try your staining again, without any antigen retieval and an overnight incubation of the primary.

If the above protocolvariation does not prove to be successful, then Ican send you another PrP antibody to try out. We have many PrP antibodies in out catalogue and I have placed links to a few below:

https://www.abcam.com/Prion-protein-PrP-antibody-8H4-ab61409.html
https://www.abcam.com/Prion-protein-PrP-antibody-EP1802Y-ab52604.html
https://www.abcam.com/Prion-protease-resistant-protein-27-30-antibody-ab7303.html



Please let me know if the protocol variations does not prove to be successful and if you do want to recieve a different antibody, which antibody you would like to try and also include the original order information aswell.

I look forward to your reply.

Read More

Answer

Thank you for contacting Abcam.

I am sorry that you are having problems with ab6664 in IHC-P on human sections. The antibody is covered under our Abpromise for six months and is guaranteed to work in IHC-P on human samples . If we cannot resolve the issue you are having with the antibody then I would be happy to either send a replacement antibody or to process a refund.

To help understand the protocol that you are using and to see if there are any suggestions that I can make, could you please answer the questions below:

1) Fixation step - Formic Acid

Fixation time

Fixation temperature

2) Antigen retrieval method (time and procedure)-heat unmasking, heat unmasking + proteinase K treatment and protease treatment

3) Permeabilization method:

Did you do a permeabilization step (details please) or add permeabilizing agent in any dilution buffers?

Permeabilizing agent and concentration:

4) Blocking agent (eg BSA, serum…):

Concentration

Blocking time

Blocking temperature

5) Endogenous peroxidases blocked?

Endogenous biotins blocked?

6) Primary antibody (If more than one was used, describe in “additional notes”) :

Concentration or dilution

Diluent buffer

Incubation time

7) Secondary antibody:

Species:

Reacts against:

Concentration or dilution

Diluent buffer

Incubation time

Fluorochrome or enzyme conjugate

8) Washing after primary and secondary antibodies:

Buffer

Number of washes

9) Detection method

10) How many times have you run this staining?

Do you obtain the same results every time?

What steps have you altered to try and optimize the use of this antibody?

If we cannot determine why the antibody is not working as it should then I will be happy to help find a replacement antibody for you.

I look forward to your reply.

Read More

Answer

ab6664 has been shown to react with mouse PrPc via western blotting. So if detectable levels of this protein are expressed in N2a cells, then we would guarantee that ab6664 will detect it. Thus no testing agreement will be necessary. I hope this is helpful. Please contact us again if you have any further information.

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1-10 of 12 Abreviews or Q&A

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