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重组RabMAb

重组Anti-Prion蛋白PrP抗体[EP1802Y] (ab52604)

Reviews (2)Q&A (4)References (31)
Western blot - Anti-Prion protein PrP antibody [EP1802Y] (ab52604)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Prion protein PrP antibody [EP1802Y] (ab52604)
  • Flow Cytometry (Intracellular) - Anti-Prion protein PrP antibody [EP1802Y] (ab52604)
  • Western blot - Anti-Prion protein PrP antibody [EP1802Y] (ab52604)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Prion protein PrP antibody [EP1802Y] (ab52604)
  • Flow Cytometry (Intracellular) - Anti-Prion protein PrP antibody [EP1802Y] (ab52604)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Prion protein PrP antibody [EP1802Y] (ab52604)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Prion protein PrP antibody [EP1802Y] (ab52604)
  • Anti-Prion protein PrP antibody [EP1802Y] (ab52604)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EP1802Y] to Prion protein PrP
  • Suitable for: Flow Cyt (Intra), WB, IHC-P
  • Reacts with: Mouse, Rat, Human

Conjugates logo Related conjugates and formulations

Alexa Fluor® 555 Alexa Fluor® 594 Alexa Fluor® 647 Alkaline Phosphatase APC Carrier Free HRP PE

概述

  • 产品名称

    Anti-Prion蛋白PrP抗体[EP1802Y]
    参阅全部 Prion protein PrP 一抗

  • 描述

    兔单克隆抗体[EP1802Y] to Prion蛋白PrP

  • 宿主

    Rabbit

  • 经测试应用

    适用于: Flow Cyt (Intra), WB, IHC-Pmore details
    不适用于: ICC

  • 种属反应性

    与反应: Mouse, Rat, Human

  • 免疫原

    Synthetic peptide within Human Prion protein PrP aa 200-300 (C terminal). The exact sequence is proprietary.

  • 阳性对照

    • WB: Human, rat and mouse brain tissue lysates. SK-MEL-28 and Neuro-2a whole cell lysates. ab74056; IHC-P: brain glioma tissue, Human, mouse and rat cerebrum tissue; Flow Cyt (intra): SH-SY5Y cells.

  • 常规说明

    This product is a recombinant monoclonal antibody, which offers several advantages including:

    • - High batch-to-batch consistency and reproducibility
    • - Improved sensitivity and specificity
    • - Long-term security of supply
    • - Animal-free production
    For more information see here.

    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.

性能

应用

The Abpromise guarantee

Abpromise™承诺保证使用ab52604于以下的经测试应用

“应用说明”部分 下显示的仅为推荐的起始稀释度；实际最佳的稀释度/浓度应由使用者检定。

应用 Ab评论 说明
Flow Cyt (Intra)
1/70.

For unpurified use at 1/00.  ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.
WB (2)
1/5000 - 1/10000. Detects a band of approximately 28 kDa (predicted molecular weight: 28 kDa).
IHC-P
1/500. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

See IHC antigen retrieval protocols.
说明
Flow Cyt (Intra)
1/70.

For unpurified use at 1/00.  ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.
WB
1/5000 - 1/10000. Detects a band of approximately 28 kDa (predicted molecular weight: 28 kDa).
IHC-P
1/500. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

See IHC antigen retrieval protocols.
应用说明
Is unsuitable for ICC.

靶标

  • 功能

    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).

  • 疾病相关

    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.

  • 序列相似性

    Belongs to the prion family.

  • 结构域

    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.

  • 翻译后修饰

    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.

  • 细胞定位

    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

  • 数据库链接

    see all

  • 别名

    • 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

图片

  • Western blot - Anti-Prion protein PrP antibody [EP1802Y] (ab52604)
    Western blot - Anti-Prion protein PrP antibody [EP1802Y] (ab52604)
    All lanes : Anti-Prion protein PrP antibody [EP1802Y] (ab52604) at 1/5000 dilution (Purified)

    Lane 1 : Human brain lysate
    Lane 2 : Mouse brain lysate
    Lane 3 : Rat brain lysate
    Lane 4 : SK-MEL-28 (Human malignant melanoma ) whole cell lysate
    Lane 5 : Neuro-2a (Mouse neuroblastoma neuroblast) whole cell lysate

    Lysates/proteins at 20 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG (HRP) with minimal cross-reactivity with human IgG at 1/2000 dilution

    Predicted band size: 28 kDa
    Observed band size: 20-37 kDa why is the actual band size different from the predicted?



    The molecular weights observed represent different glycosation states and are consistent with what has been described in the literature (PMID: 20670940, PMID: 19568430, PMID: 15240877 and PMID: 22558368).

    Blocking/Diluting buffer: 5% NFDM/TBST

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Prion protein PrP antibody [EP1802Y] (ab52604)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Prion protein PrP antibody [EP1802Y] (ab52604)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human cerebrum tissue sections labeling Prion protein PrP with purified ab52604 at 1/500 dilution (1.32 µg/mL). Heat mediated antigen retrieval was performed using Bond™ Epitope Retrieval Solution 1 (pH 6.0). Rabbit specific IHC polymer detection kit HRP/DAB (ab209101) was used as the secondary antibody. Negative control: PBS instead of the primary antibody. Hematoxylin was used as a counterstain.
    The immunostaining was performed on a Leica Biosystems BOND® RX instrument.
  • Flow Cytometry (Intracellular) - Anti-Prion protein PrP antibody [EP1802Y] (ab52604)
    Flow Cytometry (Intracellular) - Anti-Prion protein PrP antibody [EP1802Y] (ab52604)

    Intracellular Flow Cytometry analysis of SH-SY5Y (Human neuroblastoma epithelial cell) cells, labeling Prion protein PrP with Purified ab52604 at 1/70 dilution (10µg/ml) (Red). Cells were fixed with 4% Paraformaldehyde and permeabilised with 90% Methanol. A Goat anti rabbit IgG (Alexa Fluorr® 488, ab150077) secondary antibody was used at 1/2000. Isotype control - Rabbit monoclonal IgG (Black). Unlabeled control - Cell without incubation with primary antibody and secondary antibody (Blue). 

     

     

  • Western blot - Anti-Prion protein PrP antibody [EP1802Y] (ab52604)
    Western blot - Anti-Prion protein PrP antibody [EP1802Y] (ab52604)
    Anti-Prion protein PrP antibody [EP1802Y] (ab52604) at 1/5000 dilution (unpurified) + Recombinant Mouse Prion protein PrP (ab74056) at 0.01 µg

    Secondary
    Goat Anti-Rabbit IgG H&L (HRP) preadsorbed (ab97080) at 1/5000 dilution

    Developed using the ECL technique.

    Performed under reducing conditions.

    Predicted band size: 28 kDa


    Exposure time: 10 seconds


    This blot was produced using a 4-12% Bis-tris gel under the MES buffer system. The gel was run at 200V for 35 minutes before being transferred onto a Nitrocellulose membrane at 30V for 70 minutes. The membrane was then blocked for an hour using 2% Bovine Serum Albumin before being incubated with ab52604 overnight at 4°C. Antibody binding was detected using an anti-rabbit antibody conjugated to HRP, and visualised using ECL development solution ab133406.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Prion protein PrP antibody [EP1802Y] (ab52604)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Prion protein PrP antibody [EP1802Y] (ab52604)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of rat cerebrum tissue sections labeling Prion protein PrP with purified ab52604 at 1/500 dilution (1.32 µg/mL). Heat mediated antigen retrieval was performed using Bond™ Epitope Retrieval Solution 1 (pH 6.0). Rabbit specific IHC polymer detection kit HRP/DAB (ab209101) was used as the secondary antibody. Negative control: PBS instead of the primary antibody. Hematoxylin was used as a counterstain.
    The immunostaining was performed on a Leica Biosystems BOND® RX instrument.
  • Flow Cytometry (Intracellular) - Anti-Prion protein PrP antibody [EP1802Y] (ab52604)
    Flow Cytometry (Intracellular) - Anti-Prion protein PrP antibody [EP1802Y] (ab52604)

    Overlay histogram showing SH-SY5Y cells stained with unpurifiedab52604 (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 (ab52604, 1/100 dilution) for 30 min at 22°C. The secondary antibody used was DyLight® 488 goat anti-rabbit IgG (H+L) (ab96899) at 1/500 dilution for 30 min at 22°C. Isotype control antibody (black line) was rabbit IgG (monoclonal) (1µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed. 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. 

     

     

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Prion protein PrP antibody [EP1802Y] (ab52604)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Prion protein PrP antibody [EP1802Y] (ab52604)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of mouse cerebrum tissue sections labeling Prion protein PrP with purified ab52604 at 1/500 dilution (1.32 µg/mL). Heat mediated antigen retrieval was performed using Bond™ Epitope Retrieval Solution 1 (pH 6.0). Rabbit specific IHC polymer detection kit HRP/DAB (ab209101) was used as the secondary antibody. Negative control: PBS instead of the primary antibody. Hematoxylin was used as a counterstain.
    The immunostaining was performed on a Leica Biosystems BOND® RX instrument.
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Prion protein PrP antibody [EP1802Y] (ab52604)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Prion protein PrP antibody [EP1802Y] (ab52604)

    Immunohistochemical analysis of brain glioma using ab52604 (unpurified) at a dilution of 1/100. Heat mediated antigen retrieval was performed with citrate buffer pH 6 before commencing with IHC staining protocol.

  • Anti-Prion protein PrP antibody [EP1802Y] (ab52604)
    Anti-Prion protein PrP antibody [EP1802Y] (ab52604)

实验方案

To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.

Click here to view the general protocols

文献 (31)

发表研究结果有使用 ab52604？请让我们知道，以便我们可以引用本数据表中的参考文章。

ab52604 被引用在 31 文献中.

  • Mortberg MA  et al. Regional variability and genotypic and pharmacodynamic effects on PrP concentration in the CNS. JCI Insight 7:N/A (2022). PubMed: 35133987
  • Hirata T  et al. Loss of the N-acetylgalactosamine side chain of the GPI-anchor impairs bone formation and brain functions and accelerates the prion disease pathology. J Biol Chem 298:101720 (2022). PubMed: 35151686
  • Hirata T  et al. ER entry pathway and glycosylation of GPI-anchored proteins are determined by N-terminal signal sequence and C-terminal GPI-attachment sequence. J Biol Chem 298:102444 (2022). PubMed: 36055406
  • Luo Q  et al. Prion Protein Expression is Correlated with Glioma Grades. Virol Sin 35:490-493 (2020). PubMed: 32236816
  • Leca I  et al. A proteomic survey of microtubule-associated proteins in a R402H TUBA1A mutant mouse. PLoS Genet 16:e1009104 (2020). PubMed: 33137126
View all Publications for this product

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1-6 of 6 Abreviews or Q&A

Western blot abreview for Anti-Prion protein PrP antibody [EP1802Y]

 Good
Abreviews
abreview image
Application
Western blot
Sample
Mouse Cell lysate - whole cell (Neuro-2A)
Gel Running Conditions
Reduced Denaturing (4-20% gel)
Loading amount
15 µg
Treatment
Synucleozid 0, 0.25, 0.5 1uM for 48hrs
Specification
Neuro-2A
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 24°C
Read More
The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

Abcam user community

Verified customer

提交于 Apr 12 2021

Western blot abreview for Anti-Prion protein PrP antibody [EP1802Y]

 Inconclusive
Abreviews
abreview image
Application
Western blot
Sample
Mouse Cell lysate - whole cell (Neuroblasome cells)
Gel Running Conditions
Non-reduced Non-Denaturing (Native)
Loading amount
100 µg
Specification
Neuroblasome cells
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 20°C
Read More
The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

Imane Ghafir El Idrissi

Verified customer

提交于 Feb 12 2016

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
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

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
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

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
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.

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Question

  Dear Sir/Madam I would like to order your prion like protein (ab753). However, I need a primary antibody to detect it. Can you please confirm that your antibody (ab52604) is able to detect that prion like protein. I look forward to hearing from you.

Read More
Answer

Thank you for contacting us. The immunogen used for ab52604 was a smaller peptide from human Prion protein, whereas the protein ab753 is recombinant bovine Prion protein. Ab52604 was not tested for bovine and is not guaranteed to react; a better choice for use with ab753 is ab703, which used ab753 as the immunogen and will definately detect it. I hope this information is helpful to you. Please do not hesitate to contact us if you need any more advice or information.  

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