Anti-CBR1抗体(ab4148)

Reviews (1)Q&A (4)References (17)
Western blot - Anti-CBR1 antibody (ab4148)
  • Immunocytochemistry/ Immunofluorescence - Anti-CBR1 antibody (ab4148)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CBR1 antibody (ab4148)
  • Western blot - Anti-CBR1 antibody (ab4148)

Key features and details

  • Goat polyclonal to CBR1
  • Suitable for: ICC/IF, WB, IHC-P
  • Reacts with: Human
  • Isotype: IgG

重组抗体的批间可重复性更强

Product image
Anti-CBR1 antibody [EPR9660] (ab156590)
  • 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-CBR1抗体
    参阅全部 CBR1 一抗

  • 描述

    山羊多克隆抗体to CBR1

  • 宿主

    Goat

  • 经测试应用

    适用于: ICC/IF, WB, IHC-Pmore details

  • 种属反应性

    与反应: Human

  • 免疫原

    Synthetic peptide corresponding to Human CBR1 aa 265-277 (C terminal).
    Sequence:

    C-HGQFVSEKRVEQW


    (Peptide available as ab22991)
    Run BLAST with BLAST the sequence with ExPASy Run BLAST with BLAST the sequence with NCBI

  • 阳性对照

    • WB: U251 cell lysate; IHC-P: Human kidney tissue; ICC/IF: U251 cells.

  • 常规说明

    Principal Names - CBR1; CBR; carbonyl reductase 1; carbonyl reductase (NADPH); carbonyl reductase (NADPH) 1 Official Gene Symbol - CBR1 GenBank Accession Number – NP_001748 LocusLink ID - 873 (human) Gene Ontology terms - carbonyl reductase (NADPH) activity; metabolism; cytosol; oxidoreductase activity.

    Reproducibility is key to advancing scientific discovery and accelerating scientists’ next breakthrough.

    Abcam is leading the way with our range of recombinant antibodies, knockout-validated antibodies and knockout cell lines, all of which support improved reproducibility.

    We are also planning to innovate the way in which we present recommended applications and species on our product datasheets, so that only applications & species that have been tested in our own labs, our suppliers or by selected trusted collaborators are covered by our Abpromise™ guarantee.

    In preparation for this, we have started to update the applications & species that this product is Abpromise guaranteed for.

    We are also updating the applications & species that this product has been “predicted to work with,” however this information is not covered by our Abpromise guarantee.

    Applications & species from publications and Abreviews that have not been tested in our own labs or in those of our suppliers are not covered by the Abpromise guarantee.

    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, as well as customer reviews and Q&As.

性能

应用

Our Abpromise guarantee covers the use of ab4148 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
ICC/IF Use a concentration of 10 µg/ml.
WB Use a concentration of 0.03 - 0.1 µg/ml. Detects a band of approximately 30-35 kDa (predicted molecular weight: 30 kDa).

Primary incubation for 1 hour at room temperature.
IHC-P Use a concentration of 3 - 5 µg/ml. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

靶标

  • 功能

    NADPH-dependent reductase with broad substrate specificity. Catalyzes the reduction of a wide variety of carbonyl compounds including quinones, prostaglandins, menadione, plus various xenobiotics. Catalyzes the reduction of the antitumor anthracyclines doxorubicin and daunorubicin to the cardiotoxic compounds doxorubicinol and daunorubicinol. Can convert prostaglandin E2 to prostaglandin F2-alpha. Can bind glutathione, which explains its higher affinity for glutathione-conjugated substrates. Catalyzes the reduction of S-nitrosoglutathione.

  • 序列相似性

    Belongs to the short-chain dehydrogenases/reductases (SDR) family.

  • 细胞定位

    Cytoplasm.
  • Information by UniProt

  • 数据库链接

    • 别名

      • 15 hydroxyprostaglandin dehydrogenase [NADP+] antibody
      • 15-hydroxyprostaglandin dehydrogenase [NADP+] antibody
      • Carbonyl reductase [NADPH] 1 antibody
      • Carbonyl Reductase 1 antibody
      • CBR 1 antibody
      • CBR1 antibody
      • CBR1_HUMAN antibody
      • CRN antibody
      • NADPH dependent carbonyl reductase 1 antibody
      • NADPH-dependent carbonyl reductase 1 antibody
      • Prostaglandin 9 ketoreductase antibody
      • Prostaglandin 9-ketoreductase antibody
      • Prostaglandin E(2) 9 reductase antibody
      • Prostaglandin-E(2) 9-reductase antibody
      • SDR21C1 antibody
      see all

    图片

    • Western blot - Anti-CBR1 antibody (ab4148)
      Western blot - Anti-CBR1 antibody (ab4148)
      Anti-CBR1 antibody (ab4148) at 0.03 µg/ml + Human liver lysate at 35 µg

      Predicted band size: 30 kDa



      Primary incubation 1 hour at room temperature. Detected by chemiluminescence.

    • Immunocytochemistry/ Immunofluorescence - Anti-CBR1 antibody (ab4148)
      Immunocytochemistry/ Immunofluorescence - Anti-CBR1 antibody (ab4148)

      Immunocytochemistry/Immunofluorescence analysis of U251 cells labelling CBR1 with ab4148 at 10 µg/mL. Cells were paraformaldehyde fixed and permeabilized with 0.15% triton. Primary antibody incubation was for 1 hour followed by incubation with an Alexa Fluor® 488-conjugated secondary antibody at 2 µg/mL. DAPI nuclear counterstain was used.

      Negative control: Unimmunized goat IgG (10 µg/mL) followed by an Alexa Fluor® 488-conjugated secondary antibody (2 µg/mL).

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CBR1 antibody (ab4148)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CBR1 antibody (ab4148)

      ab4148 (3ug/ml) staining human CBR1 in human kidney by immunohistochemistry using paraffin embedded tissue.

      Microwaved antigen retrieval with citrate buffer pH 6, HRP-staining.

    • Western blot - Anti-CBR1 antibody (ab4148)
      Western blot - Anti-CBR1 antibody (ab4148)This image is courtesy of an anonymous Abreview
      Lanes 1 & 10 : Protein Marker
      Lanes 2-9 : Anti-CBR1 antibody (ab4148) at 1/1000 dilution (in PBS + (0.05%) Tween at 25°C )

      Lanes 1 & 10 : Protein Marker
      Lanes 2-5 : Whole cell lysate of human lung cancer cell line A549 at 25 µg
      Lane 6 : 1X SDS Loading Buffer
      Lane 7 : recombinant CBR1 at 0.03 µg
      Lane 8 : recombinant CBR1 at 0.07 µg
      Lane 9 : recombinant CBR1 at 0.1 µg

      Secondary
      Lanes 2-9 : An HRP-conjugated anti-goat IgG at 1/10000 dilution

      Developed using the ECL technique.

      Predicted band size: 30 kDa
      Observed band size: 30 kDa


      Exposure time: 1 minute


      Blocking Step: 5% Milk for 1 hour at 25°C

    文献 (17)

    发表研究结果有使用 ab4148？请让我们知道，以便我们可以引用本数据表中的参考文章。

    ab4148 被引用在 17 文献中.

    • Kubícek V  et al. Carbonyl Reduction of Flubendazole in the Human Liver: Strict Stereospecificity, Sex Difference, Low Risk of Drug Interactions. Front Pharmacol 10:600 (2019). PubMed: 31191322
    • Šadibolová M  et al. Sesquiterpenes Are Agonists of the Pregnane X Receptor but Do Not Induce the Expression of Phase I Drug-Metabolizing Enzymes in the Human Liver. Int J Mol Sci 20:N/A (2019). PubMed: 31540101
    • Kajimura T  et al. Overexpression of carbonyl reductase 1 inhibits malignant behaviors and epithelial mesenchymal transition by suppressing TGF-ß signaling in uterine leiomyosarcoma cells. Oncol Lett 18:1503-1512 (2019). PubMed: 31423217
    • Zhou S  et al. RACK1 promotes hepatocellular carcinoma cell survival via CBR1 by suppressing TNF-a-induced ROS generation. Oncol Lett 12:5303-5308 (2016). PubMed: 28105239
    • Phillips RJ  et al. Prostaglandin pathway gene expression in human placenta, amnion and choriodecidua is differentially affected by preterm and term labour and by uterine inflammation. BMC Pregnancy Childbirth 14:241 (2014). IHC . PubMed: 25048443
    View all Publications for this product

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    Western blot abreview for Anti-CBR1 antibody

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    Abreviews
    abreview image
    Application
    Western blot
    Sample
    Human Cell lysate - whole cell (Lung cancer cell line (A549))
    Loading amount
    25 µg
    Specification
    Lung cancer cell line (A549)
    Gel Running Conditions
    Reduced Denaturing (12% Gel)
    Blocking step
    Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
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    Abcam user community

    Verified customer

    提交于 Oct 10 2011

    Question

    A customer has submitted a technical enquiry regarding products related to AB4148 (CBR1 antibody). Question: The customer enquired about the recommended positive controls for this antibody - human liver lysates. A refined search of human liver lysates returned the following link listing 72 lysates: https://www.abcam.com/index.html?t=63167&pt=12&c=3998 [https://www.abcam.com/index.html?t=63167&pt=12&c=3998] Which of these 72 lysates might be most suitable for use as a positive control?

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    Answer

    Thank you for your enquiry. We do not seem to have an exact match where the lysate has been extracted in RIPA buffer. My understanding is that the protein is cytoplasmic, therefore a lysate extracted in a gentler buffer should be ok. I would recommend looking at the following product: ab29889 https://www.abcam.com/index.html?datasheet=29889 I hope this information helps. Please do not hesitate to contact us if you need any more advice or information.

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    Question

    Our customer has the problem with the antibody ab4148 in WB experiment, please check the protocol and figures.THANK YOU! 1. Order details: a.. Batch number: 321914 b.. Abcam order or Purchase order number: ab4148 c.. Antibody storage conditions (temperature/reconstitution etc) 2. Please describe the problem (high background, wrong band size, more bands, no band etc). High background and wrong band size 3. On what material are you testing the antibody in WB? · Species: human · Cell extract or Nuclear extract: all · Purified protein or Recombinant protein: purified protein 3. The lysate a.. How much protein was loaded:40ug a.. What lysis buffer was used: RIPA buffer b.. What protease inhibitors were used: leupeptin aprotinin PMSF c.. What loading buffer was used: 5xsample buffer d.. Did you heat the samples: temperature and time:95℃ 10min 4. Electrophoresis/Gel conditions/ Transfer conditions a.. Reducing or non reducing gel: b.. Gel percentage : 10% c.. Transfer conditions: 90v 60min 5. Blocking conditions a.. Buffer: TTBS or PBS-T (T: 10% Tween 20) b.. Blocking agent: milk, BSA, serum, what percentage:5%BSA or 5%milk c.. Incubation time:overnight 4℃ d.. Incubation temperature: 4℃ 6. Primary Antibody a.. Specification (in which species was it raised against):human · At what dilution(s) have you tested this antibody: 1:1000 1:500(as show) 1:200 1:100 · What dilution buffer was used:5%bsa or 5% milk · Incubation time:4℃ overnight · Incubation temperature: 4℃ · What washing steps were done:1Xttbp or 1xpbs (5min x 3 times) 7. Secondary Antibody a.. Specification (in which species was it raised against)?Rabbit anti-Goat HRP b.. At what dilution(s) have you tested this antibody: 1:3000~1:5000 。 This second antibody has been tried for many times a.. Incubation time 1-2hr in room temp b.. Wash steps: 15min x 3 times c.. Do you know whether the problems you are experiencing come from the secondary? impossible 8. Detection method ECl, ECl+, other detection method: ECL 9. Background bands · Have you eliminated the possibility that any background bands could be due to the secondary antibody? (Run a “No primary” control): NO · Is the blocking step sufficient? YES · Are your washing steps sufficiently stringent? (Multiple short washes are more effective than fewer longer wash steps) YES · At what size are the bands migrating? Could they be degradation products of your target? 30kb · Please provide an image of your blot (as an e-mail attachment, a faxed image is not sufficient) 33kd 40kd 11. Did you apply positive and negative controls along with the samples? Please specify. 10. Optimization attempts · How many times have you tried the Western?3 times or more · Do you obtain the same results every time e.g. are background bands always in the same place? almost · What steps have you altered? As above

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    Answer

    Thank you for your enquiry. I am sorry to hear that you are experiencing difficulties with this product ab4148 in western blot. Often it is possible to make suggestions that help resolve problems. We will happily offer technical support and in the event that a product is not functioning in the applications cited on the product data sheet (and the problem has been reported within 120 days of purchase), and if it appears that the antibody is at fault, a replacement/credit note/refund will be offered. I have looked through the protocols you used and have a few questions and suggestions that might help you resolve the problem. If the suggestions do not prove to be helpful, would you please be so kind to confirm the following items in order to help me better understand the cause of the problem. In most cases, the cause of high background is because the primary and/or secondary antibody concentration is too high and causes non-specific binding. The dilution that we have on the datasheet is a recommended starting dilution and customers are encouraged to determine the optimal concentration/dilution. You can try decreasing the primary (1:2000 ~ 1:5000) and secondary antibody concentration. Another possible cause is that the amount of protein is too much. We normally recommend using 20-40 micrograms per well but please try 20ug if you have not already done so. Increasing the washing frequency and duration would also help eliminate high background. Can you confirm that you have used Tween in your washing solution and have washed the primary antibody at least 15min x 3 times? Please try this if you have not already done so. Can you confirm you have reduced and denatured the sample in buffer containing SDS and mercaptoethanol? This will ensure the protein is in the correct conformation to run at the correct molecular weight and be detected by the antibody. I am sorry for the string of questions but I hope the above recommendations may already help you. If you have already tried the above suggestions and still experience problems, please do not hesitate to contact me with the answers to the above. Also, please advice me on how you would like to proceed with your enquiry, so that I can immediately arrange for a replacement or refund to you.

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    Question

    Our customer has the problem with the antibody ab4148 in WB experiment,　please check the protocol and figures.THANK YOU! 1. Order details: a.. Batch number: 321914 b.. Abcam order or Purchase order number: ab4148 c.. Antibody storage conditions (temperature/reconstitution etc) 2. Please describe the problem (high background, wrong band size, more bands, no band etc). High background and wrong band size 3. On what material are you testing the antibody in WB? · Species: human · Cell extract or Nuclear extract:all · Purified protein or Recombinant protein:purified protein 3. The lysate a.. How much protein was loaded:80ug a.. What lysis buffer was used: RIPA buffer b.. What protease inhibitors were used: luepeptein aprotenin c.. What loading buffer was used: 5xsample buffer d.. Did you heat the samples: temperature and time:95℃ 5min 4. Electrophoresis/Gel conditions/ Transfer conditions a.. Reducing or non reducing gel: b.. Gel percentage : 10% c.. Transfer conditions: 90v 60min 5. Blocking conditions a.. Buffer: TTBS or PBS b.. Blocking agent: milk, BSA, serum, what percentage:5%BSA or 5%milk c.. Incubation time:overnight 4℃ d.. Incubation temperature: 4℃ 6. Primary Antibody a.. Specification (in which species was it raised against):human · At what dilution(s) have you tested this antibody: 1:1000 1:500(as show) 1:200 1:100 · What dilution buffer was used:5%bsa or 5% milk · Incubation time:4℃ overnight · Incubation temperature: 4℃ · What washing steps were done:1Xttbp or 1xpbs 3 times 5min 7. Secondary Antibody a.. Specification (in which species was it raised against)?Rabbit anti-Goat HRP b.. At what dilution(s) have you tested this antibody: many times c.. Incubation time 1-2hr in roomtemp d.. Wash steps: 3 times 5min e.. Do you know whether the problems you are experiencing come from the secondary? impossible 8. Detection method ECl, ECl+, other detection method: ECL 9. Background bands · Have you eliminated the possibility that any background bands could be due to the secondary antibody? (Run a “No primary” control): NO · Is the blocking step sufficient? YES · Are your washing steps sufficiently stringent? (Multiple short washes are more effective than fewer longer wash steps) YES · At what size are the bands migrating? Could they be degradation products of your target? 30kb · Please provide an image of your blot (as an e-mail attachment, a faxed image is not sufficient) 31kd 24kd 40kd 55kd 11. Did you apply positive and negative controls along with the samples? Please specify. 10. Optimization attempts · How many times have you tried the Western?5 times or more · Do you obtain the same results every time e.g. are background bands always in the same place? almost · What steps have you altered?

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    Answer

    Thank you for your enquiry. I am sorry to hear that you are experiencing difficulties with this product ab4148 in western blot. Often it is possible to make suggestions that help resolve problems. We will happily offer technical support and in the event that a product is not functioning in the applications cited on the product data sheet (and the problem has been reported within 120 days of purchase), and if it appears that the antibody is at fault, a replacement/credit note/refund will be offered. I have looked through the protocols you used and have a few questions and suggestions that might help you resolve the problem. In most cases, the cause of multiple bands is because the amount of protein is too much. We normally recommend using 20-40 micrograms per well. Loading more than this may overload the gel, which will result in many non-specific bands. Please try this amount if you have not already done so. Another possible cause is that the primary and/or secondary antibody concentration is too high and causes non-specific binding. The dilution that we have on the datasheet is a recommended starting dilution and customers are encouraged to determine the optimal concentration/dilution. You can try decreasing the primary (1:2000 ~ 1:5000) and secondary antibody concentration or run a no-primary control (without the primary antibody). Running a no-primary control will be able to eliminate the possibility that your secondary antibody is binding non-specifically. Regarding the bands that you obtained, the bands located above the expected molecular weight band could be multimers. Please try boiling your protein in SDS-PAGE for 10 minutes rather than 5 minutes to ensure proper disruption of multimers. This will ensure the protein is in the correct conformation to run at the correct molecular weight and be detected by the antibody. Meanwhile, bands located below the expected molecular weight may indicate that your target protein has been digested. Please make sure that you incorporated sufficient protease inhibitors and have kept your samples on ice the whole time. Increasing the washing frequency and duration would also help eliminate high background. Can you confirm that you have used Tween in your washing solution and have washed at least 15min x 3 times? I hope the above recommendations may already help you. If you still experience problems please do not hesitate to contact me with the answers to the above questions. Also, please advice on how you would to proceed with this enquiry so that I can immediately arrange for a replacement or refund to you.

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    Question

    I would like to use the antibodies ab4148 in IHC, however I am not sure which second antibodies I should buy from Abcam, especially that ab4148 were tested only for Western blot. Do you think that ab 6740 are going to work with mentioned product? If you can help me in selection of second antibodies for IHC (with biotin) of either paraffin or frozen sections, I would be grateful.

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    Answer

    Yes I would recommend your using ab6740 (Rabbit polyclonal to Goat IgG H&L (biotin)) to detect ab4148. This secondary antibody works very well in IHC - please see the two customer reviews accessible via the online datasheet. As you have seen, ab4148 has only been tested in WB and we do not yet know if it will work in IHC. Should you go ahead and test this antibody, please let us know how you get on.

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