Anti-ICAM1 抗体 [15.2] (ab20)

Reviews (1)Q&A (6)References (17)

製品の概要

  • 製品名
    Anti-ICAM1 antibody [15.2]
    ICAM1 一次抗体 製品一覧
  • 製品の詳細
    Mouse monoclonal [15.2] to ICAM1
  • 由来種
    Mouse
  • アプリケーション
    適用あり: WB, Inhibition Assay, Flow Cyt, IHC-Fr, IPmore details
    適用なし: IHC-P
  • 種交差性
    交差種: Human
  • 免疫原

    Human monocytes.

  • ポジティブ･コントロール
    • WB: SK N BE, Saos-2 and HT1080 whole cell lysate. Flow Cytometry: HK-2 cells stimulated with TGF-beta1. IHC: Human amniotic membrane tissues.

製品の特性

アプリケーション

Our Abpromise guarantee covers the use of ab20 in the following tested applications.

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

アプリケーション Abreviews 特記事項
WB Use at an assay dependent concentration. Predicted molecular weight: 58 kDa.
Inhibition Assay Use at an assay dependent concentration. PubMed: 23825944
Flow Cyt Use at an assay dependent concentration.

ab170190 - Mouse monoclonal IgG1, is suitable for use as an isotype control with this antibody.
IHC-Fr Use at an assay dependent concentration.
IP Use at an assay dependent concentration.
  • 追加情報
    Is unsuitable for IHC-P.

    • ターゲット情報

    • 機能
      ICAM proteins are ligands for the leukocyte adhesion protein LFA-1 (integrin alpha-L/beta-2). During leukocyte trans-endothelial migration, ICAM1 engagement promotes the assembly of endothelial apical cups through ARHGEF26/SGEF and RHOG activation. In case of rhinovirus infection acts as a cellular receptor for the virus.
    • 配列類似性
      Belongs to the immunoglobulin superfamily. ICAM family.
      Contains 5 Ig-like C2-type (immunoglobulin-like) domains.
    • 翻訳後修飾
      Monoubiquitinated, which is promoted by MARCH9 and leads to endocytosis.
    • 細胞内局在
      Membrane.
    • Information by UniProt
    • 参照データベース
      • 別名
        • Antigen identified by monoclonal antibody BB2 antibody
        • BB 2 antibody
        • BB2 antibody
        • CD 54 antibody
        • CD_antigen=CD54 antibody
        • CD54 antibody
        • Cell surface glycoprotein P3.58 antibody
        • Human rhinovirus receptor antibody
        • ICAM 1 antibody
        • ICAM-1 antibody
        • ICAM1 antibody
        • ICAM1_HUMAN antibody
        • intercellular adhesion molecule 1 (CD54), human rhinovirus receptor antibody
        • Intercellular adhesion molecule 1 antibody
        • Major group rhinovirus receptor antibody
        • MALA 2 antibody
        • MALA2 antibody
        • MyD 10 antibody
        • MyD10 antibody
        • P3.58 antibody
        • Surface antigen of activated B cells, BB2 antibody
        see all

      画像

      • Western blot - Anti-ICAM1 antibody [15.2] (ab20)
        Western blot - Anti-ICAM1 antibody [15.2] (ab20)
        All lanes : Anti-ICAM1 antibody [15.2] (ab20) at 5 µg/ml

        Lane 1 : SK N BE (Human neuroblastoma) whole cell lysate
        Lane 2 : Saos-2 (Human epithelial-like osteosarcoma cell line) whole cell lysate
        Lane 3 : HT1080 (Human fibrosarcoma) whole cell lysate

        Lysates/proteins at 10 µg per lane.

        Secondary
        All lanes : Goat polyclonal to Mouse IgG - H&L - Pre-Adsorbed (HRP) at 1/3000 dilution

        Predicted band size: 58 kDa
        Observed band size: 58 kDa

      • Immunohistochemistry (Frozen sections) - Anti-ICAM1 antibody [15.2] (ab20)
        Immunohistochemistry (Frozen sections) - Anti-ICAM1 antibody [15.2] (ab20)Allen et al. PLoS One. 2013 Oct 30;8(10):e78441. doi: 10.1371/journal.pone.0078441. eCollection 2013. Fig 2.

        Expression of proteins with functions involved in ocular disease and wound healing in AM substrates.

        Proteins detected include growth factors and biomarkers (A) cell adhesion, cytokine and angiogenesis markers (B) metalloproteases and (C) neurotrophic factors (D). The collective pattern in staining demonstrates comparable levels of expression in trehalose and raffinose treated AM compared to fresh and increased expression compared to cryopreserved AM. Positive staining is represented as green or yellow and AEC nuclei were counterstained with DAPI (blue). Images shown are representative of triplicate experiments carried out on three donor membranes. Scale bar, 100 µm.

      • Flow Cytometry - Anti-ICAM1 antibody [15.2] (ab20)
        Flow Cytometry - Anti-ICAM1 antibody [15.2] (ab20)Morishita et al. PLoS One. 2011;6(8):e23267. doi: 10.1371/journal.pone.0023267. Epub 2011 Aug 5. Fig 1.

        ICAM-1 expression on HK-2 cells.

        HK-2 cells (Mock) and HK-2 cells stimulated with TGF-β1 (10.0, 1.0 and 0.1 ng/ml) were stained with ab20 and isotype control mAb (A). HK-2 cells (Mock) and HK-2 cells stimulated with TGF-β1 (10.0 ng/ml) were stained with ab20 and isotype control mAb at 24 hrs, 48 hrs and 72 hrs after TGF-β1 stimulation (B). Cells were analyzed by flow cytometry. Closed histogram: ICAM-1-stained cells. Dotted line: isotype control stained cells. MFI: mean fluorescence intensity.

      • Inhibition Assay - Anti-ICAM1 antibody [15.2] (ab20)
        Inhibition Assay - Anti-ICAM1 antibody [15.2] (ab20)Avril et al. PLoS Pathog. 2013;9(6):e1003430. doi: 10.1371/journal.ppat.1003430. Epub 2013 Jun 27. Fig S2.

        Binding levels of P. falciparum-infected erythrocytes increase after endothelial cell selection.

      参考文献

      This product has been referenced in:
      • van Balen P  et al. Tissue Damage Caused by Myeloablative, but Not Non-Myeloablative, Conditioning before Allogeneic Stem Cell Transplantation Results in Dermal Macrophage Recruitment without Active T-Cell Interaction. Front Immunol 9:331 (2018). Read more (PubMed: 29535719) »
      • Wang M  et al. Intercellular adhesion molecule 1 antibody-mediated mesoporous drug delivery system for targeted treatment of triple-negative breast cancer. J Colloid Interface Sci 538:630-637 (2018). Read more (PubMed: 30554096) »
      See all 17 Publications for this product

      ab20 を使用した論文を発表された方は、こちらまでお知らせください。データシートに掲載させていただきます。

      レビューと Q&A

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      1-7 of 7 Abreviews or Q&A

      Question

      I am getting no signal on my Western blots with your antibody used at a final concentration of 5 ug / mL in 5% milk - 1x TBST buffer. I do not even get background signal. I have tried lower and higher concentration of protein as well as primary antibodies. I am using an ectocervical cell line (ect-1) that expresses the mRNA for ICAM-1. I have never used this product before. My samples were prepared in 2% SDS-50mM Tris-HCl pH 7.5 and separately have tried lysis with RIPA buffer. I include 2x HALT protease inhibitor cocktail in the lysis. I also boil my samples in loading buffer that does not have DTT or B-ME. Any help is appreciated.

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      Answer

      Thank you for contacting us. I am sorry to hear that this antibody is not providing satisfactory results.

      The details provided will enable us to investigate this case and will provide us with vital information for monitoring product quality. Having reviewed this case, I would like to offer some suggestions to help optimize the results from ab20.

      You should use a reducing agent like DTT of b-ME. The antibody has been used to detect protein treated in reduced and denatured conditions.

      How much protein are you loading? We recommend at least 30 ug.

      Also you should try incubating the primary antibody incubation overnight at 4C if you aren't already.

      What dilution have you tried the anti-mouse secondary antibody? Has the secondary worked well with other mouse primary antibodies?

      Have you tried a loading control on the same membranes to verify transfer?

      Should the suggestions not improve the results, please do let me know.

      In the event that a product is not functioning in the species and applications cited on the product datasheet (and the problem has been reported within 6 months of purchase), we would be pleased to provide a free of charge replacement, credit note, or refund.

      I hope this information is helpful, and I thank you for your cooperation.

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      Question

      We have informed that the product ab27536, which was discontinued. We have been received the information given by you, but I contacted with the customer and she told me she need the antibody is able to recognize the ICAM1 protein intra- and extracellular in samples extracted from human. I would like you may confirm these antibodies recommended by you could work in this condition. Additionally, in the Abcam webpage, I found three antibodies, which are shown below: Anti-ICAM1 antibody [MEM-111] ab2213 Anti-ICAM1 antibody [YN1/1.7.4] ab25375 Anti-ICAM1 antibody (Biotin) ab7815 Which could these ones the differences with the others? Thanks for your support

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      Answer

      Thank you for the response.

      Any monoclonal antibody is produced by a single immune cell, which has been cloned. These will only recognize a single epitope of the target protein. This epitope typically is 5-10 amino acids in length. Therefore any single monoclonal antibody will only recognize a region within a single domain and not stain both the intracellular and extracellular region of the target protein.

      Polyclonal antibodies are created by many different immune cells and will contain multiple epitope recognition sites. Although the amount of antibody specific to a particular epitope will vary from lot-to-lot, it is possible for a polyclonal to therefore contain antibodies which will recognize different domains.

      I have reviewed the antibodies which you have listed as well as other polyclonals in our catalog to try and find a product which might be able to recognize the intracellular and extracellular regions.

      ab2213Anti-ICAM1 antibody [MEM-111]: This monoclonal antibody was created with an immunogen from a whole tissue / cell preparation (Human). We have not mapped the exact location of the epitope. Therefore the epitope recognized might be intracellular, transmembrane or extracellular.

      ab25375Anti-ICAM1 antibody [YN1/1.7.4]: This product is also monoclonal.The details of the immunogen for this antibody are not available. However as a monoclonal, this product will not be able to recognize sites in multiple domains.

      ab7815Anti-ICAM1 antibody (Biotin): This product is a rabbit polyclonal and has been raised against the whole protein. As such it is possible that the product will contain antibodies against both extracellular and intracellular regions. We do not test for this type of response and cannot guarantee that the productwill recognizeboth domains. Please be aware that this product is also biotin labeled.

      Our other rabbit polyclonal antibodies were each created from recombinant fragments and would also not be able to recognize both extracellular and intracellular domains.

      I hope this information is helpful. Please do not hesitate to contact me with further questions.

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      Question

      A customer in Chile was using ab27536, which we discontinued. The customer was very happy with this product and would like to know if we have a substitute to offer. He was using it in human tissue, western blot. The immunogen was a Recombinant fragment, corresponding to amino acids 253-367 of human ICAM1.

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      Answer

      While many of our ICAM1 antibodies have been tested in Western blot on human and are guaranteed to work under the terms of our Abpromise, in this instance I would recommend one of the following products. ab96485:https://www.abcam.com/ICAM1-antibody-6G12-ab96485.html Like the ab27536, this product is has been created from a recombinant protein corresponding to the extracellular portion of ICAM1. It is a mouse monoclonal ascites product which is tested in and guaranteed to work in Western blot and ELISA on human. ab20:https://www.abcam.com/ICAM1-antibody-15-2-ab20.html This product is a mouse monoclonal,Protein A purified, which has beentested inWB,Flow Cyt,IHC-Fr,IP. This product has been also been referencedin three publications. ab109361:https://www.abcam.com/ICAM1-antibody-EPR4776-ab109361.html Like ab27536 this product has been produced in rabbit. Rabbit monoclonals exhibit very high specificity and affinity. Besides being guaranteed for use in WB, this product will also work inIP,IHC-P andFlow Cyt. Each product offers the same guarantee and each would be a recommended replacement for ab27536. However each antibody does have its differences ( immunogen region, publications or rabbit affinity) which may appeal to the customer. I hope this is helpful. Please let me know if you have any questions.

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      Question

      Hello, That data sheets says "raised in Balb/c" - sorry for my ignorance. Is this rat or mouse? Thank you in advance.

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      Answer

      Balb/c is a mouse strain

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      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) abreview for Anti-ICAM1 antibody [15.2]

       Poor
      Abreviews
      Abcam has not validated the combination of species/application used in this Abreview.
      Application
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
      Sample
      Human Tissue sections (Urothelium, Bladder Cancer)
      Specification
      Urothelium, Bladder Cancer
      Fixative
      Paraformaldehyde
      Antigen retrieval step
      Heat mediated
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      Mr. Rudolf Jung

      Verified customer

      投稿 Aug 22 2006

      Question

      We're considering the antibody ab20 for use in functional studies, but we wanted to make sure that it was not in a buffer containing azide. Would you please let me know whether it does contain azide?

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      Answer

      This antibody does not contain azide

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      Question

      Are all antibodies 2952 - 2955 of the IgG1 subclass? This is important for us to chose the right secondary antibody - we are considering IgG1 PerCP from Becton Dickinson. Are you aware of a different red-shifted secondary antibody one could use with 488 laser excitation? Thanks a lot, kind regards, Ulf Steller Janssen Research Foundation Belgium

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      Answer

      Yes, all abs 2952-2955 are IgG1. Abcam has a FITC conjugated sheep anti-mouse IgG secondary, product code ab11-500, which may be suitable for your needs.

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      Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"

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