Anti-Chloramphenicol Acetyltransferase抗体[CAT-1] (ab5410)

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Key features and details

  • Mouse monoclonal [CAT-1] to Chloramphenicol Acetyltransferase
  • Suitable for: WB, ICC
  • Reacts with: Species independent
  • Isotype: IgG1

概述

  • 产品名称

    Anti-Chloramphenicol Acetyltransferase抗体[CAT-1]

  • 描述

    小鼠单克隆抗体[CAT-1] to Chloramphenicol Acetyltransferase

  • 宿主

    Mouse

  • 经测试应用

    适用于: WB, ICCmore details

  • 种属反应性

    与反应: Species independent

  • 免疫原

    Recombinant full length protein.

性能

  • 形式

    Liquid

  • 存放说明

    Shipped at 4°C. Upon delivery aliquot and store at -20°C or -80°C. Avoid repeated freeze / thaw cycles.

  • 存储溶液

    Constituent: PBS
  • Concentration information loading...

  • 纯度

    Protein A purified

  • 克隆

    单克隆

  • 克隆编号

    CAT-1

  • 骨髓瘤

    Sp2/0-Ag14

  • 同种型

    IgG1

  • 研究领域

应用

Our Abpromise guarantee covers the use of ab5410 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 a concentration of 2 µg/ml.
ICC Use at an assay dependent dilution.

靶标

  • 相关性

    CAT is used as a tag in many systems and is used in many transcription assays. The enzyme is an effector of chloramphenicol resistance in bacteria.

  • 别名

    • Cat antibody

文献 (1)

发表研究结果有使用 ab5410？请让我们知道，以便我们可以引用本数据表中的参考文章。

ab5410 被引用在 1 文献中.

  • Kuang PP  et al. Activation of elastin transcription by transforming growth factor-beta in human lung fibroblasts. Am J Physiol Lung Cell Mol Physiol 292:L944-52 (2007). WB ; Human . PubMed: 17209135

客户评价及客户问答

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1-3 of 3 Abreviews or Q&A

Question

Which isoform of CAT was this antibody raised against? There are various forms of CAT. Customer is using Type II

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Answer

Thank you for your enquiry. I am sorry for the delay to your enquiry. I have been in correspondence with the source of this antibody. I can tell you that both ab5410 and ab27220 were raised against full length proteins. Unfortunately this antibody was in turn sourced from originators external to the source of this antibody. The information as to which isoforms the antibodies were raised against is not available. I am sorry that I cannot be more assistance.

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Question

A customer is interested in ab to bacterial CAT for WB and IHC. (1) ab5410 is signed as applicable to WB and ICC. Is it predicted to work for IHC (frozen sections, formalin fixed and maybe also paraffin embedded sections) as well? (2) She wants to see exogenous (injected) recombinant CAT in mouse brain tissue. Do you have any indication that it will not cross react with other tissues? (she had another ab that recognized other tissues and tissues originated from non-injected mice negative controls and thus was useless). Any additional data will be very helpfull.

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Answer

Thank you for your enquiry. With regards to the first section of the question we are unable to state whether the product will function in IHC-Fr, however since the product functions in ICC and will recognise the native epitope I would expect the product to function in IHC-Fr. With regards to the second point. We do not have any information relating to the specificity of this antibody. If the customer wishes to use this AB in IHC on mouse tissue unless the customer is using null mice or directly biotinylate the primary Ab I would expect there to be high levels of background as the secondary Ab employed would need to be an anti mouse Ab and so would cross react to some extent with the tissue itself. If you have any additional questions then please do not hesitate to get back in touch with me.

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Question

BATCH NUMBER 31857 ORDER NUMBER 744498 DESCRIPTION OF THE PROBLEM Multiple bands in the high range, above 30kd on CAT transfected 293/FLPIN cells(We transfected ourselves. One strong band at ~30kd and strongest bands at ~50 and 55kd. Lots of fainter bands in between above and below. Antibody used at three concentrations in blotto/ PBS buffer, 1/200, 1/500 and 1/1000. 1/200 gave the most intense signal at the low end, but all showed bands at the 50-60kd range. SAMPLE The cells , 293FLPIN, were transfected with a CAT gene via a lipofectamine transfection from Invitrogen. PRIMARY ANTIBODY antibody ab5410 at 1/200, 1/500, 1/1000 in 1% blotto. Wash 3 times between steps in PBS 0.1% Tweens 20 SECONDARY ANTIBODY Secondary anti mouse HRP, Amersham, at 1/10000 in 1% blotto. Wash 3 times between steps in PBS 0.1% Tweens 20 DETECTION METHOD Develop in Supersignal West Pico Chemiluminescence from Pierce for 10 mins. Film down for 25 mins before signal visible. (Another person running their western alongside ours developed theirs for 5 secs and saw a signal. POSITIVE AND NEGATIVE CONTROLS USED Did not have a positive control and did not make a negative control. If you have a positive control lysate for this antibody, I would like to get some from you. I am having a hard time finding one. This was the 3rd Western performed using this antibody. The first two Westerns we used 1/200 concentration of the antibody, and saw few bands on 15min or greater exposures at ~50kd. We no longer have anymore of this lot of antibody. ANTIBODY STORAGE CONDITIONS Reconstitute upon arrival,aliquot. Aliquots stored at -20?C. No freeze thawing of antibody. SAMPLE PREPARATION The samples were made in a RIPA lysate buffer(2.2X10^6 cells in 100ul lysis buffer) containing Hepes, NaCl, MgCl2, EGTA, Na3VO4, Sodium Pyrophosphate, NaF, 1% Triton X 100, COMPLETE Protease inhibitors from Roche, PMSF and pNPP. The lysates are needle sheared but not boiled, and heated to 85?C in load buffer for 10 mins before loading on gel. There is definite protein transfer to our PVDF membrane. AMOUNT OF PROTEIN LOADED Loaded 25 and 50ug per lane per antibody titer ELECTROPHORESIS/GEL CONDITIONS Reducing gels Nupage 4-12%. TRANSFER AND BLOCKING CONDITIONS Transfer overnight at 15V, block in 5%blotto, 0.2%Tween-20 PBS1 hr. HOW MANY TIMES HAVE YOU TRIED THE APPLICATION? 3 HAVE YOU RUN A "NO PRIMARY" CONTROL? No DO YOU OBTAIN THE SAME RESULTS EVERY TIME? Yes WHAT STEPS HAVE YOU ALTERED? Just titrating the primary. ADDITIONAL NOTES This lot of antibody is now exhausted we are trying to decide whether to attempt with this antibody or try another companies, since we now have to purchase again.

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Answer

Thank you for your enquiry and for taking your time to fill in the on-line Questionnaire. I have been trying to find some good positive control for you but am very sorry it is very difficult to get one. As you probably know this enzyme is an effector of chloramphenicol resistance in bacteria and Abcam do not have this product at all. Reading through your protocol, we would make some suggestions/comments: 1. Try to load less material 20-30 ug/per lane, 50 ug may be too much. 2. Do you know how many copies of CAT are present in your system. 3. According to Swiss-Prot data, the expected band size of CAT is around 23,6 kDa. Is there any possibility that this tag may be post-translationally modified? Certainly, we an offer you a new vial in case the antibody has gone off during shipping and/or storage. Please do let us know if you are interested in, and confirm your Abcam Order Number and Purchase Order Number. We look forward to hearing from you soon.

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