HRP Anti-GFP抗体(ab6663)

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Western blot - HRP Anti-GFP antibody (ab6663)

    Key features and details

    • HRP Goat polyclonal to GFP
    • Suitable for: WB
    • Reacts with: Species independent
    • Conjugation: HRP
    • Isotype: IgG

    概述

    • 产品名称

      HRP Anti-GFP抗体
      参阅全部 GFP 一抗

    • 描述

      HRP山羊多克隆抗体to GFP

    • 宿主

      Goat

    • 偶联物

      HRP

    • 经测试应用

      适用于: WBmore details

    • 种属反应性

      与反应: Species independent

    • 免疫原

      Recombinant full length protein corresponding to GFP aa 1-246.
      Sequence:

      MSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLKFICTT GKLPVPWPTL VTTFSYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDGNYKTRA EVKFEGDTLV NRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKVNFKIRHN IEDGSVQLAD HYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLEFVTAAGIT HGMDELYK


      Database link: P42212
      Run BLAST with BLAST the sequence with ExPASy Run BLAST with BLAST the sequence with NCBI

    • 阳性对照

      • GFP

    • 常规说明

      Designed to detect GFP and its variants in ELISA (sandwich or capture), immunoblotting and immunoprecipitation Peroxidase conjugated anti-GFP assayed by immunoblot shows a 42 kDa band when reacted with GFP on a western blot.

      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.

    • 存储溶液

      Preservative: 0.01% Gentamicin sulphate
      Constituents: 0.42% Potassium phosphate, 0.87% Sodium chloride, 1% BSA

      Do NOT add Sodium Azide!
    • Concentration information loading...

    • 纯度

      Affinity purified

    • 纯化说明

      This product was prepared from monospecific antiserum by immunoaffinity chromatography using Green Fluorescent Protein (Aequorea victoria) coupled to agarose beads followed by solid phase adsorption(s) to remove any unwanted reactivities.

    • Primary antibody说明

      Designed to detect GFP and its variants in ELISA (sandwich or capture), immunoblotting and immunoprecipitation Peroxidase conjugated anti-GFP assayed by immunoblot shows a 42 kDa band when reacted with GFP on a western blot.

    • 克隆

      多克隆

    • 同种型

      IgG

    • 研究领域

    应用

    The Abpromise guarantee

    Abpromise™承诺保证使用ab6663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
    1/2000 - 1/5000.
    说明
    WB
    1/2000 - 1/5000.

    靶标

    • 相关性

      Function: Energy-transfer acceptor. Its role is to transduce the blue chemiluminescence of the protein aequorin into green fluorescent light by energy transfer. Fluoresces in vivo upon receiving energy from the Ca2+ -activated photoprotein aequorin.

      Subunit structure: Monomer.

      Tissue specificity: Photocytes.

      Post-translational modification: Contains a chromophore consisting of modified amino acid residues. The chromophore is formed by autocatalytic backbone condensation between Ser-65 and Gly-67, and oxidation of Tyr-66 to didehydrotyrosine. Maturation of the chromophore requires nothing other than molecular oxygen.

      Biotechnological use: Green fluorescent protein has been engineered to produce a vast number of variously colored mutants, fusion proteins, and biosensors. Fluorescent proteins and its mutated allelic forms, blue, cyan and yellow have become a useful and ubiquitous tool for making chimeric proteins, where they function as a fluorescent protein tag. Typically they tolerate N- and C-terminal fusion to a broad variety of proteins. They have been expressed in most known cell types and are used as a noninvasive fluorescent marker in living cells and organisms. They enable a wide range of applications where they have functioned as a cell lineage tracer, reporter of gene expression, or as a measure of protein-protein interactions. Can also be used as a molecular thermometer, allowing accurate temperature measurements in fluids. The measurement process relies on the detection of the blinking of GFP using fluorescence correlation spectroscopy.

      Sequence similarities: Belongs to the GFP family.

      Biophysicochemical properties: Absorption: Abs(max)=395 nm
      Exhibits a smaller absorbance peak at 470 nm. The fluorescence emission spectrum peaks at 509 nm with a shoulder at 540 nm.

    • 别名

      • GFP antibody
      • Green fluorescent protein antibody

    图片

    • Western blot - HRP Anti-GFP antibody (ab6663)
      Western blot - HRP Anti-GFP antibody (ab6663)
      HRP Anti-GFP antibody (ab6663) + GFP

    文献 (34)

    发表研究结果有使用 ab6663？请让我们知道，以便我们可以引用本数据表中的参考文章。

    ab6663 被引用在 34 文献中.

    • Fashe M  et al. Ser100-Phosphorylated RORa Orchestrates CAR and HNF4a to Form Active Chromatin Complex in Response to Phenobarbital to Regulate Induction of CYP2B6. Mol Pharmacol 97:191-201 (2020). PubMed: 31924695
    • Yi M  et al. Nuclear receptor CAR-ERa signaling regulates the estrogen sulfotransferase gene in the liver. Sci Rep 10:5001 (2020). PubMed: 32193417
    • Couturier L  et al. Regulation of Notch output dynamics via specific E(spl)-HLH factors during bristle patterning in Drosophila. Nat Commun 10:3486 (2019). PubMed: 31375669
    • Bouressam ML  et al. S-nitrosoglutathione inhibits cerebrovascular angiotensin II-dependent and -independent AT1 receptor responses: A possible role of S-nitrosation. Br J Pharmacol 176:2049-2062 (2019). PubMed: 30822355
    • Rogers C  et al. Gasdermin pores permeabilize mitochondria to augment caspase-3 activation during apoptosis and inflammasome activation. Nat Commun 10:1689 (2019). PubMed: 30976076
    View all Publications for this product

    客户评价及客户问答

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    1-8 of 8 Abreviews or Q&A

    Question

    Please let me know the exact molecular weight of the antibody. Is the antibody only the Fab fragment? What is the expected affinity between the antibody and GFP?

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    Answer

    Thank you for your enquiry. To answer your questions: Purified GFP is a 27 kDa monomer consisting of 238 amino acids. Ab6663 GFP antibody is whole IgG molecule. Unfortunately we do not have details of the affinities between ab6663 and its antigen.

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    Question

    About how many WB can I do with 1 vial? (= what is the initial volume and which dilution should I use?) Same question for ab 6673

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    Answer

    The number of Western Blots will depend on the volume of diluent and dilution used. I would suggest trying a preliminary experiment doing a dilution curve with a range of concentrations (1:500-1:5000 for example) and optimising from there. If you use 10 ml of buffer and 1:500 dilution you will get 5 blots from the vial, at 1:5000 you will get 50.

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    Question

    My GFP blot comes out w/a ton of background. I am using anti-GFP-ab6663 at 1:2000 in PBS/0.1%tween-20. Any suggestions?

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    Answer

    At this point I would suggest the following to help decrease the amount of background you are seeing. Try blocking with 3% BSA. Incubate with ab6663 for 1 hour at RT rather than overnight at 4C. Also, increase the number of your washes and you may want to decrease the concentration of the primary even more (try 1:2500).

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    Question

    What was the source of GFP plates used for the ELISA?

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    Answer

    Our Elisa plates are usually from Thermo Electron. We have our in-house recipe of diluent.

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    Question

    Customer is interested in using this antibody in a sandwich ELISA and would like to know what concentration to use.

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    Answer

    Although this antibody is not tested in ELISA, I would recommend using a concentration of 1-10 ug/mL.

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    Question

    I was planning on using ab6663 to detect GFP+ cells in a paraffin-embedded organ (5um cut) from a transplanted mouse. i'll using DAB as a chromogen. i've had success using other biotinylated primary antibodies followed by HRP-Avidin then DAB. I was wondering if anyone else has used this product for this application and what concentration of ab6663 could be used. Thank you.

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    Answer

    To our knowledge, this antibody has yet to be tested in this application. All tested applications are specified on Abcam product datasheets. If you decide to go ahead and purchase this product, please let us know how you get on and in return we will forward a reward of your choice, typically an Amazon gift voucher.

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    Question

    I was wondering if AB6663 is conjugated like a secondary antibody. If so, how can I use it in a sandwich ELISA? With what other antibody?

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    Answer

    ab6663 is not conjugated to permit direct detection in a sandwich ELISA. You must use an anti goat conjugated secondary to detect this polyclonal goat primary antibody.

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    Question

    Which of the three options for Abcam anti-GFP antibodies, rabbit, goat, mouse, which has the highest affinity? Thanks, Ann Vernallis

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    Answer

    We do not have specific affinity constant values for these antibodies but the one that is used most routinely and is most recommended is ab290.

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