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Anti-Alpha Dystroglycan抗体[2238] (ab106110)

Submit a review Q&A (5)References (5)
Western blot - Anti-Alpha Dystroglycan antibody [2238] (ab106110)

    Key features and details

    • Mouse monoclonal [2238] to Alpha Dystroglycan
    • Suitable for: WB
    • Reacts with: Mouse, Rat, Human
    • Isotype: IgG2b

    概述

    • 产品名称

      Anti-Alpha Dystroglycan抗体[2238]
      参阅全部 Alpha Dystroglycan 一抗

    • 描述

      小鼠单克隆抗体[2238] to Alpha Dystroglycan

    • 宿主

      Mouse

    • 特异性

      ab106110 is specific for a glycoepitope present on brain Alpha Dystroglycan. The clone was generated using enriched bovine brain Alpha dystroglycan, and has been shown to bind Alpha Dystroglycan from enriched mouse, rat, rabbit and bovine brain tissues but did not bind Alpha Dystroglycan from striated muscle, peripheral nerve or kidney (PMID: 16709410).

    • 经测试应用

      适用于: WBmore details

    • 种属反应性

      与反应: Mouse, Rat, Human

    • 免疫原

      Full length native protein (purified) corresponding to Cow Alpha Dystroglycan. Purified bovine brain alpha-dystroglycan
      Database link: O18738

    • 表位

      Glycoepitope on brain bovine alpha-dystroglycan

    • 阳性对照

      • WB: Human brain tissue lysate; SH-SY5Y cell lysate; Mouse brain tissue lysate; Rat brain tissue lysate.

    • 常规说明

      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

    应用

    The Abpromise guarantee

    Abpromise™承诺保证使用ab106110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
    WB
    1/100. Predicted molecular weight: 97 kDa.
    说明
    WB
    1/100. Predicted molecular weight: 97 kDa.

    靶标

    • 功能

      The dystroglycan complex is involved in a number of processes including laminin and basement membrane assembly, sarcolemmal stability, cell survival, peripheral nerve myelination, nodal structure, cell migration, and epithelial polarization.
      Alpha-dystroglycan is an extracellular peripheral glycoprotein that acts as a receptor for both extracellular matrix proteins containing laminin-G domains, and for certain adenoviruses. Receptor for laminin-2 (LAMA2) and agrin in peripheral nerve Schwann cells. Also acts as a receptor for M.leprae in peripheral nerve Schwann cells but only in the presence of the G-domain of LAMA2, and for lymphocytic choriomeningitis virus, Old World Lassa fever virus, and clade C New World arenaviruses.
      Beta-dystroglycan is a transmembrane protein that plays important roles in connecting the extracellular matrix to the cytoskeleton. Acts as a cell adhesion receptor in both muscle and non-muscle tissues. Receptor for both DMD and UTRN and, through these interactions, scaffolds axin to the cytoskeleton. Also functions in cell adhesion-mediated signaling and implicated in cell polarity.

    • 组织特异性

      Expressed in a variety of fetal and adult tissues. In epidermal tissue, located to the basement membrane. Also expressed in keratinocytes and fibroblasts.

    • 疾病相关

      Defects in DAG1 are the cause of muscular dystrophy-dystroglycanopathy limb-girdle type C7 (MDDGC7) [MIM:613818]. An autosomal recessive muscular dystrophy showing onset in early childhood, and associated with mental retardation without structural brain anomalies. Note=MDDGC7 is caused by DAG1 mutations that interfere with normal post-translational processing, resulting in defective DAG1 glycosylation and impaired interactions with extracellular-matrix components. Other muscular dystrophy-dystroglycanopathies are caused by defects in enzymes involved in protein O-glycosylation.

    • 序列相似性

      Contains 1 peptidase S72 domain.

    • 翻译后修饰

      O- and N-glycosylated. Alpha-dystroglycan is heavily O-glycosylated comprising of up to two thirds of its mass and the carbohydrate composition differs depending on tissue type. Mucin-type O-glycosylation is important for ligand binding activity. O-mannosylation of alpha-DAG1 is found in high abundance in both brain and muscle where the most abundant glycan is Sia-alpha-2-3-Gal-beta-1-4-Glc-NAc-beta-1-2-Man. In muscle, glycosylation on Thr-379 by a phosphorylated O-mannosyl glycan with the structure 2-(N-acetylamido)-2-deoxygalactosyl-beta-1,3-2-(N-acetylamido)-2-deoxyglucosyl-beta-1,4-6-phosphomannose is mediated by like-acetylglucosaminyltransferase (LARGE) protein and is required for laminin binding. O-mannosylation is also required for binding lymphocytic choriomeningitis virus, Old World Lassa fever virus, and clade C New World arenaviruses. The O-glycosyl hexose on Thr-367, Thr-369, Thr-372, Thr-381 and Thr-388 is probably mannose. O-glycosylated in the N-terminal region with a core 1 or possibly core 8 glycan. The beta subunit is N-glycosylated.
      Autolytic cleavage produces the alpha and beta subunits. In cutaneous cells, as well as in certain pathological conditions, shedding of beta-dystroglcan can occur releasing a peptide of about 30 kDa.
      SRC-mediated phosphorylation of the PPXY motif of the beta subunit recruits SH2 domain-containing proteins, but inhibits binding to WWW domain-containing proteins, DMD and UTRN. This phosphorylation also inhibits nuclear entry.

    • 细胞定位

      Secreted > extracellular space and Cell membrane. Cytoplasm > cytoskeleton. Nucleus > nucleoplasm. The monomeric form translocates to the nucleus via the action of importins and depends on RAN. Nuclear transport is inhibited by Tyr-892 phosphorylation. In skeletal muscle, this phosphorylated form locates to a vesicular internal membrane compartment. In peripheral nerves, localizes to the Schwann cell membrane. Colocalizes with ERM proteins in Schwann-cell microvilli.
    • Information by UniProt

    • 数据库链接

      • 别名

        • 156DAG antibody
        • A3a antibody
        • AGRNR antibody
        • Alpha dystroglycan antibody
        • Alpha-DG antibody
        • Beta-DG antibody
        • Beta-dystroglycan antibody
        • DAG antibody
        • Dag1 antibody
        • DAG1_HUMAN antibody
        • Dystroglycan 1 (dystrophin associated glycoprotein 1) antibody
        • Dystroglycan antibody
        • Dystrophin associated glycoprotein 1 antibody
        • Dystrophin-associated glycoprotein 1 antibody
        • OTTHUMP00000210857 antibody
        • OTTHUMP00000210858 antibody
        see all

      图片

      • Western blot - Anti-Alpha Dystroglycan antibody [2238] (ab106110)
        Western blot - Anti-Alpha Dystroglycan antibody [2238] (ab106110)
        All lanes : Anti-Alpha Dystroglycan antibody [2238] (ab106110) at 1/100 dilution

        Lane 1 : Human brain tissue lysate
        Lane 2 : SH-SY5Y cell lysate
        Lane 3 : Mouse brain tissue lysate
        Lane 4 : Mouse heart tissue lysate
        Lane 5 : Rat brain tissue lysate
        Lane 6 : Rat heart tissue lysate

        Lysates/proteins at 20 µg per lane.

        Performed under reducing conditions.

        Predicted band size: 97 kDa
        Observed band size: 75-100 kDa why is the actual band size different from the predicted?



        Lanes 1 - 6: Merged signal (red and green). Green - ab106110 observed at 75-100 kDa. Red - loading control ab181602 (Rabbit Anti-GAPDH antibody [EPR16891]) observed at 37 kDa.

        ab106110 was shown to react with DAG1 in Western blot. Membranes were blocked in fluorescent western blot (TBS-based) blocking solution before incubation with ab106110 and ab181602 (Rabbit Anti-GAPDH antibody [EPR16891]) overnight at 4 °C at a 1 in 100 dilution and a 1 in 20000 dilution respectively. Blots were incubated with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 h at room temperature before imaging.

      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 (5)

      发表研究结果有使用 ab106110？请让我们知道，以便我们可以引用本数据表中的参考文章。

      ab106110 被引用在 5 文献中.

      • Day BW  et al. The dystroglycan receptor maintains glioma stem cells in the vascular niche. Acta Neuropathol 138:1033-1052 (2019). PubMed: 31463571
      • Ko SF  et al. Inducible pluripotent stem cell-derived mesenchymal stem cell therapy effectively protected kidney from acute ischemia-reperfusion injury. Am J Transl Res 10:3053-3067 (2018). PubMed: 30416650
      • Barik A  et al. LRP4 is critical for neuromuscular junction maintenance. J Neurosci 34:13892-905 (2014). WB ; Mouse . PubMed: 25319686
      • Meilleur KG  et al. Clinical, pathologic, and mutational spectrum of dystroglycanopathy caused by LARGE mutations. J Neuropathol Exp Neurol 73:425-41 (2014). PubMed: 24709677
      • D'Arcy CE  et al. Identification of FHL1 as a therapeutic target for Duchenne muscular dystrophy. Hum Mol Genet N/A:N/A (2013). PubMed: 24087791

      客户评价及客户问答

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      1-5 of 5 Abreviews or Q&A

      Question

      Antibody : Anti-alpha dystroglycan antibody (2238)
      Abcam no : ab106110
      lot No. : GR45713-4
      Purchase order no : SGRH-PO/GMHLA/56/11-12 dated 23.11.11
      date of receiving : 3.12.11
      date of opening the vial : 26.5.12
      1) Samples:
      Sample is human muscle biopsy sections (skeletal muscle)
      2) Secondary antibody/detection:
      I am using NOVOLINK polymer detection kit from NOVOCASTRA. i have been using this routinely for other primary antibodies but used it for the first time for abcam antibody.
      3) Positive control: normal muscle sample was used as positive control, but no staining was observed.
      Please solve the problem asap, as my patientsare waiting for the results.

      Read More
      Answer

      Thank you for your email. We had 4 days of holidays, which is why your inquiry wasn't dealt in time. On the other hand Dr Tanya is out of office this week so I will be dealing with her inquiries.

      I have read the whole protocol and information you have provided. It seems that you haven't fixed the samples; we always recommend customer fixing the samples in ice cold fixative e.g. acetone, methanol or ethanol for 15 minutes; this is necessary in IHC-FoFr which helps to avoid the antigen retrieval method.

      The specificity note of ab106110 datasheet states, the antibody is only specific for a glycoepitope on brain bovine Alpha Dystroglycan, which is absent on Alpha Dystroglycan expressed in all other tissues. So we recommend using brain samples as positive control.

      ab64568 was developed using muscle sample this ab could be perfect with your muscle sample.

      I hope this information will be helpful. Should you have any question please do not hesitate to contact me.

      Read More

      Question

      We usually don't fix the samples for our routine IHC, and this is the first time we have procured the antibody from abcam. we are using the muscle biopsy samples not the brain sample and we are looking for alpha-dystroglycan staining on the muscle fibres for our neuromuscular dystrophy patients. is it that we have procured the wrong antibody?

      as you are suggesting that ab64568 was developed using muscle sample and this ab would be ideal for muscle sample. I request you to kindly make a replacement of the antibody that we have got with the one you are suggesting. As the muscle biopsy is an invasive procedure , two of our patients are awainitng the IHC results. Though we procured the antibody in Dec., clinically suspected patient came in May and the vial was opened in may.

      I request you to consider the case and solve the problem asap.

      Read More
      Answer

      Thanks you for your email.

      We have provided antibody ab106110 as per your order. It is clearly mentioned on the datasheet that ab64568 only detects the brain Alpha Dystroglycan because it is specific to a particular protein which is not expressed on other tissues so I would not expecting its use with muscle samples.

      We do not change antibodies if they are incorrectly ordered and are used by customers. We can however provide exchanges in case of unopened and correctly stored antibodies. This means your order is not entitle for antibody exchange. However as your patients are waiting results so I am happy making an exception this time and will provide you ab64568 as replacement. Please note this antibody has not been tested in IHC-Fr yet so the experimental conditions has to be determined by the end user.

      Please contact our Indian distributor from whom you purchased this product. They then need to contact us for agreed replacement.

      I hope this info will be helpful. Should you have any question please do not hesitate to ask.

      Read More

      Question

      I am currently investigating the antibodies against dystroglycan both alpha and beta and have seen that you have a large selection of different clones available. The antibodies with product codes mentioned above state they have not been referenced in papers yet... is this information up to date or are there still no references or evidence that these antibodies work. If this is the case it would be helpful if you could forward any information/evidence that you that these work. I would also be happy to test them if you were able to send them free of charge.

      Thank you for any assistance you can provide,

      Read More
      Answer

      Thank you for your inquiry.

      I can confirm that we do not know of any publication using the above mentioned antibodies. I would like to reassure you that we do guarantee our product for all species and applications stated on the respective datasheet.

      If you would like to use one of our products in a species or application that it has not been previously been tested in, you might be eligible for our testing discount program.

      https://www.abcam.com/collaborationdiscount

      For more information please see below:

      At Abcam, providing quality service is just as important as providing quality products. Our Abpromise® ensures that you can trust our products to perform as stated on the datasheets, or we will offer a replacement, credit, or refund.

      The Abcam Abpromise® guarantee:
      - 100% Scientific and Customer Support for any product you buy from Abcam or one of our authorized distributors.
      - We guarantee our products work in the tested species and applications stated on the datasheet.
      - We will replace or refund products not performing as stated on the datasheet if reported within 6 months of purchase.
      - We investigate any quality concerns raised by customers to ensure our catalog contains products that perform to the highest standards.

      Please note these conditions to our Abpromise®:
      - Protocol information must be provided in order for the claim to be validated.
      - Antibodies tested in recombinant samples only are not guaranteed for use on endogenous samples.
      - “Predicted to react” information on the datasheets is provided for reference only and these species are not guaranteed.
      - Fast Track antibodies are covered for ELISA against the immunizing peptide only.

      https://www.abcam.com/abpromise

      I hope this information is helpful, but please do not hesitate to contact us with further questions.

      Read More

      Question

      Thanks for the prompt reply, i have filled up the proforma that you had sent and sending you with this mail, couldn't take the pic of the sections, but just to tell you, there was NO staining of the membrane of the muscle fibres as otherwise observed staining with the antibody on the positive sample.

      Waiting for your reply

      1) Abcam product code ab106110 (Anti-alpha dystroglycan antibody 2238)



      2) Abcam order reference number or product batch number



      3) Description of the problem No staining observed in the positive sample.



      4) Sample preparation:

      Species

      Type of sample: Fresh frozen sections, perfusion fixed frozen sections, PFA/formalin fixed paraffin embedded sections, cells in culture, other: Fresh frozen sections (muscle biopsy)

      Sample preparation

      Positive control

      Negative control



      5) Fixation step NO

      Yes/No

      If yes: Fixative agent and concentration

      Fixation time

      Fixation temperature



      6) Antigen retrieval method



      7) Permeabilization method:

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

      Permeabilizing agent and concentration:





      8) Blocking agent (eg BSA, serum…): PROTEIN BLOCK (AS GIVEN IN NOVOLINK KIT FROM NOVOCASTRA)

      Concentration

      Blocking time 5 MIN.

      Blocking temperature ROOM TEMPERATURE



      9) Endogenous peroxidases blocked? PEROXIDASE BLOCK(AS GIVEN IN NOVOLINK KIT FROM NOVOCASTRA)

      Endogenous biotins blocked?



      10) Primary antibody (If more than one was used, describe in “additional notes”) : ab106110

      Concentration or dilution : (1:50) as given in the datasheet

      Diluent buffer : TBS (tris-saline buffer)

      Incubation time



      11) Secondary antibody: NOVOLINK Polymer detection kit

      Species:

      Reacts against:

      Concentration or dilution

      Diluent buffer

      Incubation time

      Fluorochrome or enzyme conjugate



      12) Washing after primary and secondary antibodies:

      Buffer : TBS BUFFER (pH- 7.5)

      Number of washes: THREE WASHES



      13) Detection method NOVOLINK Polymer detection kit (DAB)



      14) How many times have you run this staining? REGULARLY DOING WITH OTHER PRIMARY ANTIBODIES

      Do you obtain the same results every time? DONE TWICE WITH ABCAM AB (106110), SAME NEGATIVE STAINING OBTAINED

      What steps have you altered to try and optimize the use of this antibody? DIFF. DILUTIONS TRIED (1:20, 1:40)

      Document attachment: Attaching images of your IHC is strongly recommended and can greatly speed up our investigation of your problem.

      Thanks

















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      Read More
      Answer

      Thank you for providing some useful information.

      In order to be able to process your complaint, could you please confirm the batch number and the Abcam Order number (date of purchase)?

      As our Abpromise indicates, in the event that a product is not functioning in the applications/species cited on the product data sheet (and the problem has been reported within 6 months of purchase) we will happily offer a credit note/refund to the value of the product purchased.

      Alpha Dystroglycan is expressed in a variety of foetal and adult tissues. In epidermal tissue, located to the basement membrane, it is also expressed in keratinocytes and fibroblasts

      Though you have kindly provided some details, it would be much appreciated if I could get some more information which would help me identify the source of the problem.

      1) Samples:

      Could you please specify the species of the sample? Is it human? Is the muscle biopsy from heart muscle, skeletal muscle or smooth muscle?

      2) Secondary antibody/detection:

      Does the secondary antibody recognizes the primary antibody? Could you please provide further information about the detection system specificity?

      3) Positive control: any positive control used for checking the staining conditions?

      Thank you for your understanding and co-operation in this matter. I look forward to hearing from you and hope to solve this problem as soon as possible.

      Read More

      Question

      To

      The Abcam technical expert team,



      I have recently procured the Anti-alpha dystroglycal antibody (2238) from abcam andopened it for the first time on24th may'12 for IHC staining of the frozen muscle biopsy sections, but there was no staining seen with the antibody on the positive sample.

      Iused the NOVOLINK Polymer Detection System for detecting the primary antibody.

      I would like to know whether the absence of staining is due the incompatility with the secondary detection or something else, or i have got the wrong antibody instead of antibody 2238 i should have got https://www.abcam.com/Alpha-Dystroglycan-phospho-Y893-antibody-ab118551.htmlfor my purpose of staining.

      Please help me, two of my patients are waiting for the biopsy results.



      Thanks

      Read More
      Answer

      I am sorry to hear that you have been experiencing problems using this product in the application that you wish.

      In order to assess the quality of our products I would ask that you complete a brief questionnaire relating to the application used. Often it is possible to make suggestions that may help resolve problems experienced using a particular product.

      As our Abpromise indicates, in the event that a product is not functioning in the applications/species cited on the product data sheet (and the problem has been reported within 6 months of purchase) we will happily offer a credit note/refund to the value of the product purchased.

      All our customer feedback, including complaints are monitored weekly by our in house technical support team. If a product is at fault the technical support team will consider removing the product from our catalogue in order to avoid future customer inconvenience.

      Please could you provide some further details of the protocol used and complete the following form (attached as a word document). It would be much appreciated if you could attach an image to the response.

      Thank you for your understanding and co-operation in this matter. I look forward to hearing from you soon and resolving this issue as soon as possible.

      Read More

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