Overview

  • Product name
    Anti-ICAM1 antibody [15.2]
    See all ICAM1 primary antibodies
  • Description
    Mouse monoclonal [15.2] to ICAM1
  • Host species
    Mouse
  • Tested applications
    Suitable for: WB, Inhibition Assay, Flow Cyt, IHC-Fr, IPmore details
    Unsuitable for: IHC-P
  • Species reactivity
    Reacts with: Human
  • Immunogen

    Human monocytes.

  • Positive control
    • 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.

Properties

Applications

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.

Application Abreviews Notes
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.
  • Application notes
    Is unsuitable for IHC-P.
  • Target

    • Function
      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.
    • Sequence similarities
      Belongs to the immunoglobulin superfamily. ICAM family.
      Contains 5 Ig-like C2-type (immunoglobulin-like) domains.
    • Post-translational
      modifications
      Monoubiquitinated, which is promoted by MARCH9 and leads to endocytosis.
    • Cellular localization
      Membrane.
    • Information by UniProt
    • Database links
    • Alternative names
      • 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

    Images

    • 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.

    • 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.

    • 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

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

    References

    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) »
    • Yasen A  et al. Release of HIV-1 sequestered in the vesicles of oral and genital mucosal epithelial cells by epithelial-lymphocyte interaction. PLoS Pathog 13:e1006247 (2017). Read more (PubMed: 28241053) »
    See all 13 Publications for this product

    Customer reviews and Q&As

    1-7 of 7 Abreviews or Q&A

    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.

    Read More

    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.

    Read More

    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.

    Read More

    Answer

    Balb/c is a mouse strain

    Read More
    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

    Mr. Rudolf Jung

    Verified customer

    Submitted Aug 22 2006

    Answer

    This antibody does not contain azide

    Read More

    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.

    Read More

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

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