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    prion-protein-prp-antibody-ab703.pdf

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Neuroscience Neurology process Neurodegenerative disease Prions
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Anti-Prion protein PrP 抗体 (ab703)

  • Datasheet
Submit a review Q&A (5)References (3)

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Western blot - Anti-Prion protein PrP antibody (ab703)

    Key features and details

    • Rabbit polyclonal to Prion protein PrP
    • Suitable for: WB
    • Reacts with: Mouse
    • Isotype: IgG

    リコンビナント抗体で、ロット間での高い再現性を実現

    Product image
    Anti-Prion protein PrP antibody [EP1802Y] (ab52604)
    • 異なるロット間での安定した再現性
    • 容易なスケールアップ
    • 評価試験による特異性の確認済み
    • 倫理基準に準拠 - アニマル・フリーの生産

    製品の概要

    • 製品名

      Anti-Prion protein PrP antibody
      Prion protein PrP 一次抗体 製品一覧
    • 製品の詳細

      Rabbit polyclonal to Prion protein PrP
    • 由来種

      Rabbit
    • 特異性

      Recognises natural PrPc and conformationally changed PrPc (PrPsc). Recognises recombinant protein ,ab753, in western blot and ELISA.
    • アプリケーション

      適用あり: WBmore details
    • 種交差性

      交差種: Mouse
    • 免疫原

      Recombinant full length protein corresponding to Cow Prion protein PrP aa 1-250.
      Database link: P10279

    • 特記事項

      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 this with our range of recombinant monoclonal antibodies and knockout edited cell lines for gold-standard validation. Please check that this product meets your needs before purchasing.

      If you have any questions, special requirements or concerns, please send us an inquiry and/or contact our Support team ahead of purchase. Recommended alternatives for this product can be found below, along with publications, customer reviews and Q&As

    製品の特性

    • 製品の状態

      Liquid
    • 保存方法

      Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C or -80°C. Avoid freeze / thaw cycle.
    • バッファー

      Preservative: 0.02% Sodium azide
      Constituent: Whole serum
    • Concentration information loading...
    • 精製度

      Whole antiserum
    • ポリ/モノ

      ポリクローナル
    • アイソタイプ

      IgG
    • 研究分野

      • Neuroscience
      • Neurology process
      • Neurodegenerative disease
      • Prions
      • Stem Cells
      • Hematopoietic Progenitors
      • Surface Molecules
      • Neuroscience
      • Diseases

    関連製品

    • Compatible Secondaries

      • Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077)
      • Goat Anti-Rabbit IgG H&L (HRP) (ab205718)
    • Isotype control

      • Rabbit IgG, polyclonal - Isotype Control (ChIP Grade) (ab171870)
    • Positive Controls

      • Recombinant Cow Prion protein PrP (ab753)
    • Recombinant Protein

      • Recombinant Human Prion protein PrP (denatured) (ab140567)

    アプリケーション

    The Abpromise guarantee

    Abpromise保証は、 次のテスト済みアプリケーションにおけるab703の使用に適用されます

    アプリケーションノートには、推奨の開始希釈率がありますが、適切な希釈率につきましてはご検討ください。

    アプリケーション Abreviews 特記事項
    WB
    1/2000.
    特記事項
    WB
    1/2000.

    ターゲット情報

    • 機能

      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).
    • 関連疾患

      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.
    • 配列類似性

      Belongs to the prion family.
    • ドメイン

      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.
    • 翻訳後修飾

      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.
    • 細胞内局在

      Cell membrane. Golgi apparatus and Cytoplasm. Nucleus. Accumulates outside the secretory route in the cytoplasm, from where it relocates to the nucleus.
    • Target information above from: UniProt accession P04156 The UniProt Consortium
      The Universal Protein Resource (UniProt) in 2010
      Nucleic Acids Res. 38:D142-D148 (2010) .

      Information by UniProt
    • 参照データベース

      • Entrez Gene: 19122 Mouse
      • SwissProt: P04925 Mouse
      • Unigene: 648 Mouse
      • 別名

        • 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

      画像

      • Western blot - Anti-Prion protein PrP antibody (ab703)
        Western blot - Anti-Prion protein PrP antibody (ab703)This image is courtesy of Dr. M. Glatzel, Institut of Neuropathology, University Hospital of Zürich, Switzerland

        Lane 1: MW standard
        Lane 2: wild-type mice (PrPc)
        Lane 3: CDI/PrPsc (PK-)
        Lane 4: CDI/PrPsc (PK+),
        Lane 5: PrP 0/0 mice
        CDI = clinically ill CD-1 mice, PK = digested with Proteinase K.

        Lane 1: MW standard
        Lane 2: wild-type mice (PrPc)
        Lane 3: CDI/PrPsc (PK-)
        Lane 4: CDI/PrPsc (PK+),
        Lane 5: PrP 0/0 mice
        CDI = clinically ill CD-1 mice, PK = digested with Proteinase K.

      プロトコール

      • Western blot protocols

      Click here to view the general protocols

      データシートおよび資料

      • Datasheet download

        Download

      参考文献 (3)

      ab703 を使用した論文を発表された方は、こちらまでお知らせください。データシートに掲載させていただきます。

      ab703 は 3 報の論文で使用されています。

      • Besnier LS  et al. The cellular prion protein PrPc is a partner of the Wnt pathway in intestinal epithelial cells. Mol Biol Cell 26:3313-28 (2015). IF . PubMed: 26224313
      • Manich G  et al. Presence of a neo-epitope and absence of amyloid beta and tau protein in degenerative hippocampal granules of aged mice. Age (Dordr) 36:151-65 (2014). PubMed: 23867972
      • Morel E  et al. The cellular prion protein PrP is involved in the proliferation of epithelial cells and in the distribution of junction-associated proteins. PLoS ONE 3:e3000 (2008). IP ; Human, Mouse . PubMed: 18714380

      レビューと Q&A

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      レビューを送る 質問を送る

      1-5 of 5 Abreviews or Q&A

      Question

      Hi again,

      Thanks for your reply. I have attached the requested information as a Word file, and hope all the details you need are in there.

      I'll wait for your advice in the light of this information before deciding whether to take a replacement or a refund for the ab703. It would be interesting to know if you have any further insight into my result with this antibody.

      Best Wishes,

      Read More

      Abcam community

      Verified customer

      Asked on Jan 17 2012

      Answer

      Thank you for contacting us again and for the full and clear explanation of your experiments and results.


      I am sorry that the results with this antibody has not been more successful as was indicated by the previous results with this antibody. It does indeed appear that the antibody is not detecting any PrPsc whilst the 8H4 clone is picking it up. The reason for this is unclear. From the following paper it is indicated that the Chandler strain is much more susceptible to degradation than other strains, which may mean that the levels present following PK digestion are low. It also states that the monoglycosylated form is most prevalent in this strain which may expain the single band at ˜25kDa. However, as 8H4 was able to pick out the PrPsc and the single band observed with ab703 is so clean these explanations may be stretching it a bit.


      http://www.ncbi.nlm.nih.gov/pubmed/10415165


      The conditions used to obtain the Western blot presented on the datasheet of ab703 was to treat the sample with PK (at a concentration of 20 µg/mL) for 30 minutes at 37°C then loaded on the gel following heating to 99°C for 10 minutes. May I ask, how did you quench the PK reaction? The paper above used the addition of PMSF. Were the samples used for the 8H4 and ab703 exactly the same (i.e. had they both been frozen prior to detection)?


      I would say from the results obtained it may be most worthwhile to optimise the conditions using the monoclonal antibody 8H4. Whilst the bands are not clear, especially for the PrPc, this may be improved on optimisation. Concentrating on the incubation of the antibody with the membrane as well as the blocking conditions used. It may also be necessary to look into how the samples are prepared, as the protein has a tendency to aggregate which may be contributing to the higher bands observed for the PK- lanes.


      For the incubation, I would try both 1 hour at room temperature as well as at 4°C overnight (with agitation). This can quite often lead to a dramatic reduction in non-specific banding. I would also try an alternative blocking agent such as 3% BSA in TBST or 5% serum.


      If you would like a replacement with our 8H4 antibody or any of our other anti-PrP antibodies I would be happy to arrange this. I am not sure ab52604 would be an improvement as the specificity has not been assessed and the band observed by Western blotting is again indicating the detection of only one glycosylated form.Alternatively if you would prefer a refund please do let me know.


      I am sorry I have not been able to beof more help in this case.

      Read More

      Abcam Scientific Support

      Answered on Jan 17 2012

      Question

      Phone call requesting suggestions of antibodies to use with detection of PrP (both PrP C and PrP Sc). For use in Western blotting of mouse and human samples. Would like to detect PrP Sc which is protease resistance.

      Read More

      Abcam community

      Verified customer

      Asked on Jan 03 2012

      Answer

      Thank you for contacting us again regarding our antibodies directed against PrP. I am sorry that I was not in the office and able to deal with your call. I am sorry that it has taken some time to find more information relating to the antibodies we currently have to offer and which may be the most suitable for you. We have a number of antibodies which have been confirmed to recognise both the PrPc and PrPsc forms of the protein. These include: ab6664, ab703 and ab14219. I am unsure as to how you are currently performing your experiments but it seems that the normal protocol is to use proteinase K to destroy any PrPc in your sample and then any protein which is left (PrPsc) can be detected using Western blotting. As discussed over the phone, ab703 seems to be appropriate for your experiments, however I am a little concerned that the results that you have been observing have been highly non-specific and the Western blot for this antibody has shown to give quite broad bands. This antibody is covered by the Abpromise and if you do choose to buy it we would offer you all the support we can to make it perform as expected and if you were to be unhappy with the results youmay be entitled to a free of charge replacement or a refund. I would suggest considering ab52604 as this antibody has shown very clean Western blotting bands with human fetal, mouse and ratbrain. I have attached the image to this email. Although this antibody has not been tested to see if both forms of the protein are recognised, it seems likely that it would, using the same procedure of proteinase K treatment. However, as this has not beenconclusivelyconfirmed I would be willing to offer a testing discount to you. This would involve you buying this antibody, testing it with your samples to assess the specificity and once you have shared the results with us through submitting an Abreview of your results (should only take 5-10 minutes) you would be entitled to a primaryantibody of your choice from our catalogue. More information onthis offer can befound here: https://www.abcam.com/collaboratordiscount If you would be interested in this offer please let me know and I can issue you witha discount code. I am still trying togather more informationregarding theotherantibodiesinourcatalogueand will keep youupdated with any furtherrelevant information. I hope this information has been of help.If you require any further information please let me know.

      Read More

      Abcam Scientific Support

      Answered on Jan 03 2012

      Question

          Hi thanks for your prompt response. The application is western blotting. the conjugate I am not surebut highly likely is going to be HRP. But your advice on this is welcomed.

      Read More

      Abcam community

      Verified customer

      Asked on Dec 06 2011

      Answer

      Thanks for the additional information. Here is a link to an HRP-conjugated secondary antibody you could for western blotting with ab703: https://www.abcam.com/Rabbit-IgG-secondary-antibody-ab16284.html I hope this is helpful. Please contact me again if  you have any further questions.

      Read More

      Abcam Scientific Support

      Answered on Dec 06 2011

      Question

        Hello Thanks for your previous advice. Could you please also advice me what is the best secondry antibody to detect primary anti-body ab703.

      Read More

      Abcam community

      Verified customer

      Asked on Dec 05 2011

      Answer

      Thank you for your enquiry. I would be happy to help you find a secondary antibody for use with ab703. If you could let me know what application you would use the antibody for and what conjugate you would like attached to the secondary (eg. HRP, biotin, what color fluorophore) then I can recommend a product.

      Read More

      Abcam Scientific Support

      Answered on Dec 05 2011

      Question

      I would like the conditions that you used for this Western blot that you show on your website (ie: reducing or non-reducing conditions, SDS or native, dilutions, etc.)

      Read More

      Abcam community

      Verified customer

      Asked on Oct 13 2003

      Answer

      Thank you for your enquiry and your patience. I was away so I could not answer to this technical enquiry sooner. I managed to get some more information about this product: - 12% SDS polyacrylamide gel - 25-50 ug total protein are loaded - rabbit polyclonal to prion protein diluted 1:2000 - Loading buffer is: 150 mM tris-HCl, pH 6,8, 6% SDS, 0.03% bromphenol blue, 30% glycerol - sample incubation 95°C for 5 min - the membran is developed by enhanced chemilumenscence (ECL) .

      Read More

      Asdf Edo

      Abcam Scientific Support

      Answered on Oct 13 2003

      Please note: All products are "FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC PROCEDURES"
      For licensing inquiries, please contact partnerships@abcam.com

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