Recombinant
RabMAb

Recombinant Anti-ICAM1 antibody [EPR4776] (ab109361)

Submit a review Q&A (3)References (8)

Overview

  • Product name

    Anti-ICAM1 antibody [EPR4776]
    See all ICAM1 primary antibodies

  • Description

    Rabbit monoclonal [EPR4776] to ICAM1

  • Host species

    Rabbit

  • Tested applications

    Suitable for: ICC/IF, WB, IP, IHC-P, Flow Cytmore details

  • Species reactivity

    Reacts with: Human

  • Immunogen

    Recombinant fragment, corresponding to a region near the C-terminus of Human ICAM1.

  • Positive control

    • Lysates of the following cells: Raji, Ramos, A549 treated with TNF-alpha, A549, and HUVEC treated with TNF-alpha; Human kidney tissue and Human tonsil tissue.

  • General notes

    Mouse, Rat: We have preliminary internal testing data to indicate this antibody may not react with these species. Please contact us for more information.

    This product is a recombinant monoclonal antibody, which offers several advantages including:

    • - High batch-to-batch consistency and reproducibility
    • - Improved sensitivity and specificity
    • - Long-term security of supply
    • - Animal-free production
    For more information see here.

    Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.

Properties

Applications

Our Abpromise guarantee covers the use of ab109361 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
ICC/IF 1/500.
WB 1/1000 - 1/10000. Detects a band of approximately 89 kDa (predicted molecular weight: 58 kDa).
IP 1/10 - 1/100.
IHC-P 1/100 - 1/250. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
Flow Cyt 1/100 - 1/500.

ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.

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) 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 antibody
      • Surface antigen of activated B cells, BB2 antibody
      see all

    Images

    • Immunocytochemistry/ Immunofluorescence - Anti-ICAM1 antibody [EPR4776] (ab109361)
      Immunocytochemistry/ Immunofluorescence - Anti-ICAM1 antibody [EPR4776] (ab109361)

      Immunocytochemistry/ Immunofluorescence analysis of Ramos (human Burkitt's lymphoma B lymphocyte) labeling ICAM1 with ab109361 at 1/500. ab150077, AlexaFluor®488 Goat anti-Rabbit at 1/1000 was used as the secondary antibody. Cells were fixed with 4% Paraformaldehyde and permeabilised with 0.1% tritonX-100. DAPI (blue) was used as nuclear counter stain.

      Confocal image showing membranous staining on Ramos cell line

    • Western blot - Anti-ICAM1 antibody [EPR4776] (ab109361)
      Western blot - Anti-ICAM1 antibody [EPR4776] (ab109361)
      All lanes : Anti-ICAM1 antibody [EPR4776] (ab109361) at 1/1000 dilution

      Lane 1 : Raji cell lysate
      Lane 2 : Ramos cell lysate
      Lane 3 : A549 cells treated with TNF-alpha, lysate
      Lane 4 : A549 cell lysate
      Lane 5 : HUVEC cells treated with TNF-alpha, lysate

      Lysates/proteins at 10 µg per lane.

      Predicted band size: 58 kDa
      Observed band size: 89 kDa
      why is the actual band size different from the predicted?

    • Flow Cytometry - Anti-ICAM1 antibody [EPR4776] (ab109361)
      Flow Cytometry - Anti-ICAM1 antibody [EPR4776] (ab109361)
      Overlay histogram showing Ramos cells stained with ab109361 (red line). The cells were fixed with 4% paraformaldehyde (10 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab109361, 1/1000 dilution) for 30 min at 22°C. The secondary antibody used was Alexa Fluor® 488 goat anti-rabbit IgG (H+L) (ab150077) at 1/2000 dilution for 30 min at 22°C. Isotype control antibody (black line) was rabbit IgG (monoclonal) (0.1μg/1x106 cells) used under the same conditions. Unlabelled sample (blue line) was also used as a control. Acquisition of >5,000 events were collected using a 20mW Argon ion laser (488nm) and 525/30 bandpass filter. This antibody gave a positive signal in Ramos cells fixed with 80% methanol (5 min)/permeabilized with 0.1% PBS-Tween for 20 min used under the same conditions.
    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ICAM1 antibody [EPR4776] (ab109361)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ICAM1 antibody [EPR4776] (ab109361)

      ab109361, at 1/100, staining ICAM1 in Human kidney tissue by immunohistochemistry.

      Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ICAM1 antibody [EPR4776] (ab109361)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ICAM1 antibody [EPR4776] (ab109361)

      ab1091361, at 1/100, staining ICAM1 in Human tonsil tissue by immunohistochemistry.

      Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

    References

    This product has been referenced in:

    • Tengroth L  et al. Activation of Activin receptor-like kinases curbs mucosal inflammation and proliferation in chronic rhinosinusitis with nasal polyps. Sci Rep 8:1561 (2018). Read more (PubMed: 29367682) »
    • Altmann T  et al. Endothelial cell damage in idiopathic pneumonia syndrome. Bone Marrow Transplant 53:515-518 (2018). Read more (PubMed: 29335628) »
    See all 8 Publications for this product

    Publishing research using ab109361? Please let us know so that we can cite the reference in this datasheet.

    Customer reviews and Q&As

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    1-3 of 3 Abreviews or Q&A

    Question

    Customer kindly contact us as this product was not performing in IHC.

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    Answer

    Thank you for contacting us. Your credit note ID is I am sorry that this antibody did not perform as stated on the datasheet. I have asked our accounting department to issue a credit note for you, which can be redeemed against the invoice of a future order by passing it on to your purchasing department. To avoid confusion, please ensure your accounts department is aware of how the credit note is being used. If you have questions on how to use the credit note, please contact our accounting department. If payment has already been made on the original order and you wish to receive a refund, please ask your purchasing department to contact our accounting department so that we may gather the correct information needed for the refund. To avoid confusion, please ensure your accounts department is aware of how the credit note is being used. Our accounting department can be contacted by email at us.credits@abcam.com or by telephone using the information at the Contact Us link in the top right corner of our website. Please refer to the credit note ID in any correspondence with our accounting department. The credit note ID is for your reference only and does not automatically guarantee the credit. I hope this experience will not prevent you from purchasing other products from us in the future. Our Scientific Support team is always at your service, should you require further expert advice.

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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

    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.

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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.

    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.

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