Anti-beta Galactosidase抗体[DC1 4C7] (ab116)

Submit a review Q&A (13)References (5)

概述

  • 产品名称
    Anti-beta Galactosidase抗体[DC1 4C7]
    参阅全部 beta Galactosidase 一抗
  • 描述
    小鼠单克隆抗体[DC1 4C7] to beta Galactosidase
  • 宿主
    Mouse
  • 经测试应用
    适用于: WB, ELISAmore details
  • 免疫原

    beta Galactosidase EC.3.2.1.23 from E.coli.

  • 常规说明

    This monoclonal is also known as clone HD4C7 or HD 447C.

    This antibody clone is manufactured by Abcam. If you require a particular buffer formulation or a particular conjugate for your experiments, please contact orders@abcam.com or you can find further information here.

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: 0.02% Sodium azide
    Constituent: PBS
  • Concentration information loading...
  • 纯度
    Protein G purified
  • 克隆
    单克隆
  • 克隆编号
    DC1 4C7
  • 骨髓瘤
    P3-NS1/1-Ag4-1
  • 同种型
    IgG1
  • 轻链类型
    kappa
  • 研究领域

应用

Our Abpromise guarantee covers the use of ab116 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

应用 Ab评论 说明
WB Use at an assay dependent concentration.
ELISA Use at an assay dependent concentration.

靶标

  • 相关性
    Beta galactosidase is coded by a gene (lac z) in the lac operon of Escherichia coli. It is a metalloenzyme that splits lactose into glucose and galactose. It hydrolyzes terminal, non-reducing beta-D-galactose residues in beta-D-galactosides. Activation by cations seems to be substrate dependent. K+, Na+, NH4+, Rb+, Cs+ and Mn++ all activate enzyme activity based upon the substrate used.
  • 细胞定位
    Cytoplasmic
  • 别名
    • Beta D galactosidase antibody
    • Beta gal antibody
    • Beta galactosidase antibody
    • ECK0341 antibody
    • JW0335 antibody
    • Lactase antibody
    • lacZ antibody
    see all

图片

  • Western blot - Anti-beta Galactosidase antibody [DC1 4C7] (ab116)
    Western blot - Anti-beta Galactosidase antibody [DC1 4C7] (ab116)
    All lanes : Anti-beta Galactosidase antibody [DC1 4C7] (ab116) at 1 µg

    Lane 1 : Recombinant Human GLB1 protein (ab151890)
    Lane 2 : beta Galactosidase protein (ab79449)

    Lysates/proteins at 0.1 µg per lane.

    Secondary
    All lanes : Goat polyclonal to Mouse IgG - H&L - Pre-Adsorbed (HRP) (ab65485) at 1/5000 dilution

    Performed under reducing conditions.

    Exposure time: 4 minutes

实验方案

文献

This product has been referenced in:
  • Smith SJ  et al. Recapitulation of tumor heterogeneity and molecular signatures in a 3D brain cancer model with decreased sensitivity to histone deacetylase inhibition. PLoS One 7:e52335 (2012). IHC-P . Read more (PubMed: 23272238) »
  • Nassiri M  et al. Repressors NFI and NFY participate in organ-specific regulation of von Willebrand factor promoter activity in transgenic mice. Arterioscler Thromb Vasc Biol 30:1423-9 (2010). IHC-Fr . Read more (PubMed: 20431063) »
See all 5 Publications for this product

发表研究结果有使用 ab116？请让我们知道，以便我们可以引用本数据表中的参考文章。

客户评价及客户问答

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1-10 of 13 Abreviews or Q&A

Question

We have bought mouse monoclonal [DC1 4c7] to beta Galactosidase (ab116) from your company. I trasnfected mammalian cells with Positive ctrl plasmid from Invitrogen(pcDNA4.0/Hismax/LacZ) which will generate a LacZ and His-tag fusion protein. Subsequently, I used the above antibody with 4 different dilution (500x, 2000x, 5000x, 10,000x) to perform a western blot but did not see the desire band except some unspecific bands. However, Western blot with the same sample lysate using Anti-His Antibody generated a strong and specific band. Since the antibody was shipped in ambient temperature, I worry the activity of it might have decreased. If not, can i know the dilution or other specifications of this antibody when performing western blot so that i'll have better gauge in troubleshooting.

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Answer

Thank you for your enquiry. I am sorry to hear that you have been having difficulties with this antibody. We do not have a recommended dilution of this antibody. The dilution employed will depend on the level of expression of your LacZ construct. However, I would certainly expect detection of LacZ in view of your detection of His and detection of your construct using an anti-His antibody. Can you tell me whether you are detecting whole LacZ tagged with His as I would like to confirm that the epitope is present on the construct.

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Question

BATCH NUMBER 286 ORDER NUMBER -- NOT SPECIFIED -- DESCRIPTION OF THE PROBLEM No signal or weak signal in all samples SAMPLE 1. ?-gal fusion protein from engineered ES cell 2. Over-expressed ?-gal 3. Purified ?-gal protein PRIMARY ANTIBODY Abcam / Mouse / 3% BSA / 1:2000, 1000, 500 or 100 / 1 hour at RT or overnight at 4?, 4 x 5 min wash with PBS-T DETECTION METHOD ECL POSITIVE AND NEGATIVE CONTROLS USED Positive controls : Purified ?-gal protein, ?-gal transfected cell lysate Negative control : not transfected cell lysate ANTIBODY STORAGE CONDITIONS -70? / No reconstitution SAMPLE PREPARATION 1X SDS gel loading buffer AMOUNT OF PROTEIN LOADED 10 ng for purified protein, 60 ?g for lysates ELECTROPHORESIS/GEL CONDITIONS Reducing, 6% or 7% gel TRANSFER AND BLOCKING CONDITIONS 25 mM Tris base, 192 mM glycine, 20% methanol / 2 hours, 5% BSA or 10% skim milk in PBS-T (0.1% Tween-20) SECONDARY ANTIBODY Jackson InmunoResearch / Donkey / 3% BSA / 1:10000, 5000 / 1 hour at RT, 4 x 5 min wash with PBS-T HOW MANY TIMES HAVE YOU TRIED THE APPLICATION? 10 HAVE YOU RUN A "NO PRIMARY" CONTROL? No DO YOU OBTAIN THE SAME RESULTS EVERY TIME? Yes WHAT STEPS HAVE YOU ALTERED? Antibody dilution factor, incubating condition ADDITIONAL NOTES ? We want to replace not-working monoclonal ab(ab116) with polyclonal anti-?-gal antibody(Abcam, Cat#. ab616) We guarantee our samples and immunoblotting conditions because we successfully detected endogenous Bmal1 protein(our major target) from the same blots.

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Answer

Thank you for your enquiry and for submitting your technical questionnaire. I am sorry to hear that your customer has been having difficulties with this antibody. I have read their technical enquiry and they have tried some of the recommendations that I would make such as reducing the concentration of the antibody or blocking in BSA. However, I would be interested to see the blot image and information regarding whether your customer can be certain that beta-gal is being expressed to a significant level by these cells. An enzymatic detection of beta-gal by its substrate will be significantly more sensitive than detection of the protein itself by western blotting. I understand that your customer has been using a purified protein preparation of beta-gal. However, it may be that the antibody's sensitivity cannot detect levels as low as 10ng. I appreciate the patience of your customer in this matter and look forward to hearing from you.

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Question

BATCH NUMBER 286 ORDER NUMBER ? DESCRIPTION OF THE PROBLEM No staining SAMPLE Mouse embryonic brain PRIMARY ANTIBODY Ab116 1:50, 1:100 and 1:400 SECONDARY ANTIBODY Dako goat anti-mouse 1:200 (works very well for other mouse monoclonals) DETECTION METHOD ABC, DAB POSITIVE AND NEGATIVE CONTROLS USED Tissue without Beta-gal as negative, Known beta-gal positive ANTIBODY STORAGE CONDITIONS 4 degrees celsius FIXATION OF SAMPLE Paraformaldehyde, 3hr ANTIGEN RETRIEVAL Microwaving 4x5minutes BLOCKING CONDITIONS PBS 0.1% Triton X, 10% goat serum HOW MANY TIMES HAVE YOU TRIED THE APPLICATION? 2 HAVE YOU RUN A "NO PRIMARY" CONTROL? Yes DO YOU OBTAIN THE SAME RESULTS EVERY TIME? Yes WHAT STEPS HAVE YOU ALTERED? Concentration, antigen retrieval ADDITIONAL NOTES My protocol works well for most other monoclonals, I ran samples with your monoclonal to TTF1 at the same time as positive controls and they worked beautifully. I need a beta-gal ab to work with this protocol so I can do double labelling. If possible I would like a refund or to swap the antibody for one of you other monoclonals to Beta-gal.

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Answer

Thank you for your enquiry and for taking your time to fill in the on-line Questionnaire. We are very sorry to hear that you are having problem with this antibody. We would like to confirm that this antibody has not been tested for immunohistochemistry, although it is suitable for IF and ICC. So you may need to fix the sample in a more gentle way, we would advise to fix the samples for shorter time (15 to 30 min) than you would normally do with formalin-fixed tissue sections. We have another alternative antibody ab616 which has been tested for IHC both on frozen and formalin-fixed sections and it is one of our best sellers. We have also received very good feedback information from our customers and published 15 reviews on-line. We hope this information will be useful for you.

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Question

Thank you very much for your reply. I just aware that your anti-b-gal is from rabbit. However, I am searching an anti-b-Gal with non-rabbit source, because I need it for double labeling with neural progenitor markers such as Sox-2, Sox-10 and Mash-1, which I have are rabbit source. I am just wondering whether I can get a small amount of your antibodies ab116 and ab1047 to try. We will let you know results. We can buy them and more after if it works. I think some laboratories also need a non-rabbit source of the anti-b-Gal.

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Answer

Thank you for your email. We do not routinely offer free or trial sized samples for testing purposes. Our policy at Abcam is that if an antibody does not work as specified on the datasheet, we will offer a replacement or reimbursement. Should you decide to test an antibody in an application for which we do not have any information, please let us know how you get on and in return we will award you 50 points with the Abcam Loyalty Scheme which can be redeemed on a number of rewards. If you have any additional questions, please do let us know.

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Question

Thanks for your reply. To my understanding, the ab616 will be good for detecting b-GAL by immunohistochemistry at mouse brain. I shall appreciate it very much if you can provide further information. Best regards,

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Answer

Yes, ab616 has successfully been tested for application in IHC-formalin fixed paraffin embedded sections. We have had some feedback from customers who have used ab616 in IHC-P to detect beta gal in transgenic mice - you may read their comments by clicking on the reviews tab located on the online datasheet. We also have a specific IHC protocol for this antibody - the protocol can be found in the protocols tab on the online datasheet. If you have any additional questions, please do contact us again.

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Question

I am a customer of your company. Could I ask you some questions concerning the antibodies ab116 and ab1047? Do you have any data to show which can be used for immunohistochemistry in tissue. I want to buy one of them for parafomaldehyde perfused mouse brain sections, I wonder which one you would suggest is better for that purpose? By the way, is it refundable if it doesn't work? Thanks very much in advance for your help.

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Answer

Thank you for your email. Neither ab116 or ab1047 have been tested yet for application in IHC. You may want to look at ab616 which has been tested in IHC. If you decide to try ab116 or ab1047 in IHC, please let us know how you get on and in return we will award you 50 points with the Abcam Loyalty Scheme which can be redeemed on a number of rewards. Our policy at Abcam is that if an antibody does not work as specified on the datasheet, we will offer a replacement or reimbursement. However, because ab116 and ab1047 have not yet been tested in IHC we are unable to provide a guarantee. If you have any additional questions, please contact us again.

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Question

I would like to use an anti-beta-galactosidase antibody to detect a truncated beta-gal protein containing only the N terminus (the first 300 AA). Do you know if the epitope of your ab116 antibody is within this region. Thanks in advance for your time,

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Answer

The epitope of this antibody has not been mapped.

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Question

Information suitability for immunocytochemistry.

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Answer

This anitbody has not yet been tested in immunocytochemistry

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Question

Does the antibody detect ß Gal expression by eucaryotic cells in WB? Thank you.

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Answer

Yes this antibody does work in this application (although there maybe some post-translational modification of the b-gal fusion protein which may decrease the binding of the antibody)

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Question

I would like to use an anti-beta-galactosidase antibody to detect E.coli beta-gal in an ELISA. Previously I have used Gibco clone 5B88 to coat the ELISA plates at a conc. of 2.5ug/ml. Can ab116 be used in such an application?

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Answer

Yes, this antibody works in ELISA.

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