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Immunology Innate Immunity Macrophage / Inflamm.

Anti-M6PR (cation independent)抗体[MEM-238] (ab8093)

  • Datasheet
  • SDS
Reviews (3)Q&A (7)References (14)

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Flow Cytometry (Intracellular) - Anti-M6PR (cation independent) antibody [MEM-238] (ab8093)

    Key features and details

    • Mouse monoclonal [MEM-238] to M6PR (cation independent)
    • Suitable for: Flow Cyt (Intra)
    • Reacts with: Human
    • Isotype: IgG1

    Conjugates logo Related conjugates and formulations

    Alexa Fluor® 647 Biotin FITC PE

    选择批间可重复性更高的重组抗体

    Product image
    Anti-M6PR (cation independent) antibody [EPR6599] (ab124767)
    • 研究可靠 —— 各批次间结果一致且可重复
    • 长期批量供应 —— 采用重组技术,可实现快速生产
    • 首次实验即可成功 —— 经过大量验证确认了特异性
    • 符合伦理标准 —— 产品不含动物成分

    概述

    • 产品名称

      Anti-M6PR (cation independent)抗体[MEM-238]
      参阅全部 M6PR (cation independent) 一抗
    • 描述

      小鼠单克隆抗体[MEM-238] to M6PR (cation independent)
    • 宿主

      Mouse
    • 特异性

      CD222 antigen (human)
    • 经测试应用

      适用于: Flow Cyt (Intra)more details
    • 种属反应性

      与反应: Human
      预测可用于: Non human primates
    • 免疫原

      Tissue, cells or virus corresponding to Vaccinia virus M6PR (cation independent).

    • 表位

      Recognizes an epitope between domains 2 and 5.
    • 阳性对照

      • Flow Cyt (Intra): HeLa cells.
    • 常规说明

      CD222 is a 250kDa transmembrane protein with a short cytoplasmic tail containing an internalization signal. CD222 was originally identified as a receptor for IGFII and M6P-containing proteins (e.g. lysosomal hydrolases). Lysosomal enzymes are sorted to lysosomes via CD222 either from the Golgi, where the enzymes acquire M6P, or from the extracellular space. The majority of CD222 molecules (approximately 90-95%) are located intracellularly, only 5-10% is present on the cell membrane. The internalization rate seems to be enhanced by ligand induced dimerization of CD222 as well as by phosphorylation of its cytoplasmic serine. CD222 is also a receptor for TGFbeta latency associated peptide (LAP), proliferin and may bind several molecules independently of M6P, including plasminogen, CD87 or retinoic acid. It is involved in activation of latent TGFbeta [PROW].

      This product was changed from ascites to tissue culture supernatant on 24th January 2018. Please note that the dilutions may need to be adjusted accordingly. If you have any questions, please do not hesitate to contact our scientific support team.

      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

    性能

    • 形式

      Liquid
    • 存放说明

      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.
    • 存储溶液

      pH: 7.40
      Preservative: 0.097% Sodium azide
      Constituent: PBS
    • Concentration information loading...
    • 纯度

      Protein A purified
    • 纯化说明

      Purified from TCS. Purity >95% by SDS-PAGE.
    • Primary antibody说明

      CD222 is a 250kDa transmembrane protein with a short cytoplasmic tail containing an internalization signal. CD222 was originally identified as a receptor for IGFII and M6P-containing proteins (e.g. lysosomal hydrolases). Lysosomal enzymes are sorted to lysosomes via CD222 either from the Golgi, where the enzymes acquire M6P, or from the extracellular space. The majority of CD222 molecules (approximately 90-95%) are located intracellularly, only 5-10% is present on the cell membrane. The internalization rate seems to be enhanced by ligand induced dimerization of CD222 as well as by phosphorylation of its cytoplasmic serine. CD222 is also a receptor for TGFbeta latency associated peptide (LAP), proliferin and may bind several molecules independently of M6P, including plasminogen, CD87 or retinoic acid. It is involved in activation of latent TGFbeta [PROW].
    • 克隆

      单克隆
    • 克隆编号

      MEM-238
    • 同种型

      IgG1
    • 研究领域

      • Immunology
      • Innate Immunity
      • Macrophage / Inflamm.
      • Tags & Cell Markers
      • Subcellular Markers
      • Organelles
      • Endosome
      • Signal Transduction
      • Growth Factors/Hormones
      • Insulin / Insulin-like
      • Signal Transduction
      • Protein Trafficking
      • Golgi Proteins
      • Neuroscience
      • Processes

    相关产品

    • Alternative Versions

      • Alexa Fluor® 647 Anti-M6PR (cation independent) antibody [MEM-238] (ab187589)
      • Biotin Anti-M6PR (cation independent) antibody [MEM-238] (ab239242)
      • FITC Anti-M6PR (cation independent) antibody [MEM-238] (ab58841)
      • PE Anti-M6PR (cation independent) antibody [MEM-238] (ab77135)
    • Compatible Secondaries

      • Goat Anti-Mouse IgG H&L (Alexa Fluor® 488) (ab150113)
      • Goat Anti-Mouse IgG H&L (HRP) (ab205719)
      • Goat Anti-Mouse IgG H&L (DyLight® 488) preadsorbed (ab96879)
    • Conjugation kits

      • PE / R-Phycoerythrin Conjugation Kit - Lightning-Link® (ab102918)
      • APC Conjugation Kit - Lightning-Link® (ab201807)
    • Isotype control

      • Mouse IgG1, kappa monoclonal [15-6E10A7] - Isotype Control (ab170190)
      • Mouse IgG1, Kappa Monoclonal [B11/6] - Isotype Control (ab91353)

    应用

    The Abpromise guarantee

    Abpromise™承诺保证使用ab8093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)
    Use a concentration of 2 - 6 µg/ml.

    ab170190 - Mouse monoclonal IgG1, is suitable for use as an isotype control with this antibody.

    说明
    Flow Cyt (Intra)
    Use a concentration of 2 - 6 µg/ml.

    ab170190 - Mouse monoclonal IgG1, is suitable for use as an isotype control with this antibody.

    靶标

    • 功能

      Transport of phosphorylated lysosomal enzymes from the Golgi complex and the cell surface to lysosomes. Lysosomal enzymes bearing phosphomannosyl residues bind specifically to mannose-6-phosphate receptors in the Golgi apparatus and the resulting receptor-ligand complex is transported to an acidic prelyosomal compartment where the low pH mediates the dissociation of the complex. This receptor also binds IGF2. Acts as a positive regulator of T-cell coactivation, by binding DPP4.
    • 序列相似性

      Belongs to the MRL1/IGF2R family.
      Contains 1 fibronectin type-II domain.
    • 结构域

      Contains 15 repeating units of approximately 147 AA harboring four disulfide bonds each. The most highly conserved region within the repeat consists of a stretch of 13 AA that contains cysteines at both ends.
    • 细胞定位

      Lysosome membrane. Colocalized with DPP4 in internalized cytoplasmic vesicles adjacent to the cell surface.
    • Target information above from: UniProt accession P11717 The UniProt Consortium
      The Universal Protein Resource (UniProt) in 2010
      Nucleic Acids Res. 38:D142-D148 (2010) .

      Information by UniProt
    • 数据库链接

      • Entrez Gene: 3482 Human
      • Omim: 147280 Human
      • SwissProt: P11717 Human
      • Unigene: 487062 Human
      • Unigene: 673278 Human
      • 别名

        • 300 kDa mannose 6 phosphate receptor antibody
        • 300 kDa mannose 6-phosphate receptor antibody
        • Cation independent mannose 6 phosphate receptor antibody
        • Cation-independent mannose-6-phosphate receptor antibody
        • CD222 antibody
        • CD222 antigen antibody
        • CI Man 6 P receptor antibody
        • CI Man-6-P receptor antibody
        • CI MPR antibody
        • CI-M6PR antibody
        • CI-MPR antibody
        • CIMPR antibody
        • IGF 2 receptor antibody
        • IGF 2R antibody
        • IGF II receptor antibody
        • IGF-II receptor antibody
        • IGF2 receptor antibody
        • Igf2r antibody
        • Insulin like growth factor 2 receptor antibody
        • Insulin like growth factor II receptor antibody
        • Insulin-like growth factor 2 receptor antibody
        • Insulin-like growth factor II receptor antibody
        • M6P R antibody
        • M6P/IGF2 receptor antibody
        • M6P/IGF2R antibody
        • M6PR antibody
        • mannose 6 phosphate receptor antibody
        • mannose 6 phosphate receptor, cation independent antibody
        • MPR 300 antibody
        • MPR300 antibody
        • MPRI antibody
        • MPRI_HUMAN antibody
        see all

      图片

      • Flow Cytometry (Intracellular) - Anti-M6PR (cation independent) antibody [MEM-238] (ab8093)
        Flow Cytometry (Intracellular) - Anti-M6PR (cation independent) antibody [MEM-238] (ab8093)
        Overlay histogram showing HeLa cells stained with ab8093 (red line). The cells were fixed with 80% methanol (5 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 (ab8093, 1µg/1x106 cells) for 30 min at 22ºC. The secondary antibody used was DyLight® 488 goat anti-mouse IgG (H+L) (ab96879) at 1/500 dilution for 30 min at 22ºC. Isotype control antibody (black line) was mouse IgG1 [ICIGG1] (ab91353, 2µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed.

      实验方案

      • Flow cytometry protocols

      Click here to view the general protocols

      数据表及文件

      • SDS download

      • Datasheet download

        Download

      文献 (14)

      发表研究结果有使用 ab8093?请让我们知道,以便我们可以引用本数据表中的参考文章。

      ab8093 被引用在 14 文献中.

      • Besemer AS  et al. Receptor-mediated endocytosis 8 (RME-8)/DNAJC13 is a novel positive modulator of autophagy and stabilizes cellular protein homeostasis. Cell Mol Life Sci 78:645-660 (2021). PubMed: 32322926
      • Promchan K & Natarajan V Leucine zipper transcription factor-like 1 binds adaptor protein complex-1 and 2 and participates in trafficking of transferrin receptor 1. PLoS One 15:e0226298 (2020). PubMed: 31895934
      • Hirst J  et al. Role of the AP-5 adaptor protein complex in late endosome-to-Golgi retrieval. PLoS Biol 16:e2004411 (2018). WB ; Human . PubMed: 29381698
      • Stadlmann J  et al. Comparative glycoproteomics of stem cells identifies new players in ricin toxicity. Nature 549:538-542 (2017). PubMed: 28959962
      • Chen Y  et al. Segregation in the Golgi complex precedes export of endolysosomal proteins in distinct transport carriers. J Cell Biol 216:4141-4151 (2017). PubMed: 28978644
      View all Publications for this product

      客户评价及客户问答

      Show All 评价 Q&A
      提交评价 提交问题

      1-10 of 10 Abreviews or Q&A

      Immunocytochemistry/ Immunofluorescence abreview for Anti-Mannose 6 Phosphate Receptor (Cation independent) antibody [MEM-238]

      Excellent
      Abreviews
      Abreviews
      abreview image
      Application
      Immunocytochemistry/ Immunofluorescence
      Sample
      Human Cell (A549)
      Specification
      A549
      Fixative
      Paraformaldehyde
      Permeabilization
      Yes - 0.1% Triton
      Blocking step
      5% Goat Serum, 0.5% BSA, 0.5% Gelatin as blocking agent for 16 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 4°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 Jul 07 2010

      Immunocytochemistry/ Immunofluorescence abreview for Anti-Mannose 6 Phosphate Receptor (Cation independent) antibody [MEM-238]

      Excellent
      Abreviews
      Abreviews
      abreview image
      Application
      Immunocytochemistry/ Immunofluorescence
      Sample
      Human Cell (HeLa)
      Specification
      HeLa
      Fixative
      Paraformaldehyde
      Permeabilization
      Yes - Triton X-100
      Blocking step
      BSA as blocking agent for 30 minute(s) · Concentration: 2% · Temperature: 25°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 Mar 11 2010

      Immunocytochemistry/ Immunofluorescence abreview for Anti-Mannose 6 Phosphate Receptor (Cation independent) antibody [MEM-238]

      Good
      Abreviews
      Abreviews
      abreview image
      Application
      Immunocytochemistry/ Immunofluorescence
      Sample
      Human Cell (HEK cells)
      Specification
      HEK cells
      Fixative
      Paraformaldehyde
      Blocking step
      BSA as blocking agent for 40 minute(s) · Concentration: 5%
      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.

      Dr. Randal Moldrich

      Verified customer

      提交于 Mar 31 2006

      Question

      I have repeated the protocol with your suggested amendments but unfortunately have obtained the same results. I wondered if it would be possible for you to send me a vial of ab32815 to try as I see this has been used in Western blotting in published papers with images?

      Read More

      Abcam community

      Verified customer

      Asked on Mar 06 2012

      Answer

      I have issued a free of charge replacement with the order number for ab32815.

      To check the status of the order please contact our Customer Service team and reference this number.

      Should you still be experiencing difficulties, or if you have any further questions, please do not hesitate to let us know.

      I wish you the best of luck with your research.

      Read More

      Abcam Scientific Support

      回复于 Mar 06 2012

      Question

      Brief protocol:



      1. 50,000 - 150,000 Cells lysed in 20uL running buffer containing mercaptoethanol for 5min in 95C heat block (no protease inhibitor used)



      2. Samples (20uL) run on gel with protein ladder for approx 90 mins.



      3. Transferred for 2hr - very good transfer of protein marker seen



      4. Blocked for 60 mins at RT with 5% milk



      5. Incubated with primary antibody (1 in 2000 dilution) overnight at 4C



      6. Incubated with secondary antibody for 1hr at RT



      7. ECL applied for 1 min



      8. Membrane developed (images below).

      Read More

      Abcam community

      Verified customer

      Asked on Mar 02 2012

      Answer

      Thank you, I received the data this time. For a protocol modification, I suggest trying preparing a sample by heating at 60C for 10 minutes instead of 95 for 5 minutes to minimize aggregation of the trans-membrane domains. It is strange that both antibodies give you this result, and I still think there may be some issue with the sample entering the gel and/or transferring effectively, even though the standards do.

      Another modification is to remove methanol from the transfer buffer and add SDS to a final concentration of 0.1%, to encourage transfer of this large protein.

      Are you confident that the secondary antibody is not reacting with the samples? Is the secondary specific for a subclass of mouse IgG?

      These modifications are unlikley to remove the band at 70 kDa. We do have other antibodies against the receptor, so if these modifications fail, I can send a vial of something else to try, such as ab32815 or ab124767.

      Read More

      Abcam Scientific Support

      回复于 Mar 02 2012

      Question

      I have been having trouble with two Abcam IGF2R (CI-M6PR) antibodies, a b2733 and ab8093, which I have been attempting to use for Western Blotting.

      Both antibodies have failed to show a band in the expected region (approx 250Kd) but have instead shown a band at around 70Kd. I have confirmed the cell lines that I am using (CMK and K562) express IGF2R using PCR and flow cytometry. I have attached a document containing the gels run for the two proteins. I have tried each antibody 3 times now with similar results. I have run secondary only controls, which do not show this band.

      Do you have any suggestions on how to improve my results?

      Read More

      Abcam community

      Verified customer

      Asked on Mar 02 2012

      Answer

      Thank you for bringing this to our attention. I will need a few more details of your protocol. I was not able to open the file you sent.

      Can you please tell me how you prepared your lysates (did you use protease inhibitors), and how you heated the samples to denature. I am concerned that the receptor may have aggregated upon boiling and did not enter the gel effectively, or did not transfer to the blot well. Did you check for efficacy of transfer with a Ponceau Red stain or equivalent?

      How much sample did you load per lane of the gel? How did you block the membrane? What concentration of antibody did you apply to the membrane and how long did you incubate with the antibody?

      Read More

      Abcam Scientific Support

      回复于 Mar 02 2012

      Question

      Thanks, I have just tried with BSA as blocking agent and I did have more success but with also more a specific bands from ms9-II cells that overexpress M6PR. I will also try ab32815 to see if the later ab shows less a specific bands with my ms9-II cell extract. Thanks again and you have been really very helpful

      Read More

      Abcam community

      Verified customer

      Asked on Nov 25 2011

      Answer

      I'm glad that you have been seeing better results with blocking with BSA. If you are now having difficulty with non-specific bands, increasing the dilution of the primary and secondary antibody as well as adding 1 % BSA and 0.2% Tween 20 into the diluents may help. If you would like me to have a look at your protocol to see if there is any other advice I can think of then please do fill in the form and I'd be happy to try and help. Otherwise, I wish you all the best with your experiments.

      Read More

      Abcam Scientific Support

      回复于 Nov 25 2011

      Question

      Phone call enquiring about ab2733 in regards to performing western blotting as performance with ab8093 was faint.

      Read More

      Abcam community

      Verified customer

      Asked on Nov 25 2011

      Answer

      Thank you for contacting us. Having looked into the case a little I can see no reason why ab8093 should not be performing well with Western blotting or that ab2733 would be likely to perform any better. What might be helpful is if you were to share the protocol which you are currently performing and I may be able to spot some things which may help to improve the performance of ab8093. To this end I have attached a questionnaire, this should only take 5-10 minutes to fill in. It would be helpful if you could also attach an image of the blot obtained. This information will also help us to monitor the quality of our products and to determine if any further internal testing needs to be performed. If the antibody is not performing to the level we would expect with the applications and species listed on the datasheet you would be eligible for a replacement antibody or refund. We do have an antibody in our catalogue which have been more fully characterised with Western blotting and may serve as an alternative to ab8093 if necessary. Ab32815 has been used by one of our customers to detect the Mannose 6 Phosphate Receptor protein from the membrane fraction of Hela Cells using Western blotting: https://www.abcam.com/M6PR-cation-independent-antibody-ab32815.html&tab=abreviews&intabreviewid=11713 I'm sorry that I could not be of more help but hopefully with the protocol in hand we can decide if trying an alternative antibody would be worthwhile. In the meantime I am sorry for any inconvenience this has caused.

      Read More

      Abcam Scientific Support

      回复于 Nov 25 2011

      Question

      Thank you so much. Have a great week, Jeremy.

      Read More

      Abcam community

      Verified customer

      Asked on Sep 13 2011

      Answer

      I have attached an Excel file with information regarding all our current antibodies that have been tested in EM. I hope this is helpful. Please let me know if you have any further questions.

      Read More

      Abcam Scientific Support

      回复于 Sep 13 2011

      Question

      I would like to get a list of all antibodies from Abcam that have been successfully tested for EM.

      Read More

      Abcam community

      Verified customer

      Asked on Sep 09 2011

      Answer

      Thank you for your enquiry. I am working up an internal query to obtain a list of all EM-tested antibodies in our catalog. I will forward you this list as soon as possible. Thanks for your patience!

      Read More

      Abcam Scientific Support

      回复于 Sep 09 2011

      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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