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    icam1-antibody-epr4776-ab109361.pdf

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RecombinantRabMAb

Recombinant Anti-ICAM1 antibody [EPR4776] (ab109361)

  • Datasheet
  • SDS
Reviews (1)Q&A (3)References (14)

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Western blot - Anti-ICAM1 antibody [EPR4776] (ab109361)
  • Western blot - Anti-ICAM1 antibody [EPR4776] (ab109361)
  • Western blot - Anti-ICAM1 antibody [EPR4776] (ab109361)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ICAM1 antibody [EPR4776] (ab109361)
  • Flow Cytometry (Intracellular) - Anti-ICAM1 antibody [EPR4776] (ab109361)
  • Immunoprecipitation - Anti-ICAM1 antibody [EPR4776] (ab109361)
  • Immunocytochemistry/ Immunofluorescence - Anti-ICAM1 antibody [EPR4776] (ab109361)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ICAM1 antibody [EPR4776] (ab109361)
  • Anti-ICAM1 antibody [EPR4776] (ab109361)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPR4776] to ICAM1
  • Suitable for: Flow Cyt (Intra), ICC/IF, WB, IP, IHC-P
  • Reacts with: Human

Conjugates logo Related conjugates and formulations

Alexa Fluor® 488 Alexa Fluor® 647 Biotin Carrier Free

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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: Flow Cyt (Intra), ICC/IF, WB, IP, IHC-Pmore details
  • Species reactivity

    Reacts with: Human
  • Immunogen

    Recombinant fragment within Human ICAM1 aa 300-500 (C terminal). The exact sequence is proprietary.
    Database link: P05362

  • Positive control

    • IHC-P: Human lung carcinoma and tonsil tissue sections; Flow Cyt (intra): Ramos cells; ICC/IF: Raji cells; IP: Raji whole cell lysate; WB: HUVEC treated with 10ng/ml for 18 hours and Ramos whole cell lysate.
  • General notes

    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.

    We are constantly working hard to ensure we provide our customers with best in class antibodies. As a result of this work we are pleased to now offer this antibody in purified format. We are in the process of updating our datasheets. The purified format is designated 'PUR' on our product labels. If you have any questions regarding this update, please contact our Scientific Support team.

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

Properties

  • Form

    Liquid
  • Storage instructions

    Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C long term. Avoid freeze / thaw cycle.
  • Storage buffer

    pH: 7.20
    Preservative: 0.01% Sodium azide
    Constituents: 0.05% BSA, 40% Glycerol (glycerin, glycerine), 59% PBS
  • Concentration information loading...
  • Purity

    Protein A purified
  • Clonality

    Monoclonal
  • Clone number

    EPR4776
  • Isotype

    IgG
  • Research areas

    • Immunology
    • Cell Type Markers
    • CD
    • Adhesion
    • Microbiology
    • Organism
    • Virus
    • RNA Virus
    • ssRNA positive strand virus
    • SARS Coronavirus
    • Cardiovascular
    • Atherosclerosis
    • Vascular Inflammation
    • Leukocyte recruitment
    • Cell adhesion molecules
    • Stem Cells
    • Hematopoietic Progenitors
    • Lymphoid
    • B Lymphocytic Lineage
    • Stem Cells
    • Hematopoietic Progenitors
    • Lymphoid
    • T Lymphocytic Lineage
    • Immunology
    • Adaptive Immunity
    • Regulatory T Cells
    • Cardiovascular
    • Angiogenesis
    • Endothelial Cell Markers
    • Neuroscience
    • Development

Associated products

  • Alternative Versions

    • Alexa Fluor® 488 Anti-ICAM1 antibody [EPR4776] (ab214860)
    • Alexa Fluor® 647 Anti-ICAM1 antibody [EPR4776] (ab214944)
    • Biotin Anti-ICAM1 antibody [EPR4776] (ab217075)
    • Anti-ICAM1 antibody [EPR4776] - BSA and Azide free (ab226059)
  • Isotype control

    • Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730)
  • Positive Controls

    • Raji membrane extract lysate (ab30126)

Applications

The Abpromise guarantee

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
Flow Cyt (Intra)
1/70.

For unpurified use at 1/100 - 1/500.  ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.

ICC/IF
1/100.

For unpurified use at 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)
1/100 - 1/250. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

See IHC antigen retrieval protocols.

Notes
Flow Cyt (Intra)
1/70.

For unpurified use at 1/100 - 1/500.  ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.

ICC/IF
1/100.

For unpurified use at 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.

See IHC antigen retrieval protocols.

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.
  • Target information above from: UniProt accession P05362 The UniProt Consortium
    The Universal Protein Resource (UniProt) in 2010
    Nucleic Acids Res. 38:D142-D148 (2010) .

    Information by UniProt
  • Database links

    • Entrez Gene: 3383 Human
    • Omim: 147840 Human
    • SwissProt: P05362 Human
    • Unigene: 643447 Human
    • 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

    • 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 : Wild-type HeLa cell lysate
      Lane 2 : ICAM1 CRISPR-Cas9 edited HeLa cell lysate
      Lane 3 : A549 cell lysate
      Lane 4 : Ramos cell lysate

      Lysates/proteins at 20 µg per lane.

      Performed under reducing conditions.

      Predicted band size: 58 kDa
      Observed band size: 90 kDa why is the actual band size different from the predicted?



      False colour image of Western blot: Anti-ICAM1 antibody [EPR4776] staining at 1/1000 dilution, shown in green; Mouse anti-Alpha Tubulin [DM1A] (ab7291) loading control staining at 1/20000 dilution, shown in red. In Western blot, ab109361 was shown to bind specifically to ICAM1. A band was observed at 90 kDa in wild-type HeLa cell lysates with no signal observed at this size in Icam1 CRISPR-Cas9 edited cell line ab261742 (CRISPR-Cas9 edited cell lysate ab256947). The band observed in the CRISPR-Cas9 edited lysate lane below 90 kDa is likely to represent a truncated form of ICAM1. This has not been investigated further and the functional properties of the gene product have not been determined. To generate this image, wild-type and Icam1 CRISPR-Cas9 edited HeLa cell lysates were analysed. First, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 3 % milk in TBS-0.1 % Tween® 20 (TBS-T) before incubation with primary antibodies overnight at 4 °C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged. Secondary antibodies used were Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed (ab216776) at 1/20000 dilution.

    • Western blot - Anti-ICAM1 antibody [EPR4776] (ab109361)
      Western blot - Anti-ICAM1 antibody [EPR4776] (ab109361)
      Lane 1 : Anti-ICAM1 antibody [EPR4776] (ab109361) at 1/1000 dilution (Purified)
      Lane 2 : Anti-ICAM1 antibody [EPR4776] (ab109361) at 1/1000 dilution ((Purified))

      Lane 1 : HUVEC (Human umbilical vein endothelial cell) whole cell lysate
      Lane 2 : HUVEC (Human umbilical vein endothelial cell) treated with 10ng/ml for 18 hours whole cell lysate

      Lysates/proteins at 15 µg per lane.

      Secondary
      All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution (Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated)

      Predicted band size: 58 kDa

    • Western blot - Anti-ICAM1 antibody [EPR4776] (ab109361)
      Western blot - Anti-ICAM1 antibody [EPR4776] (ab109361)
      Anti-ICAM1 antibody [EPR4776] (ab109361) at 1/1000 dilution (Purified) + Ramos (Human Burkitt's lymphoma B lymphocyte) whole cell lysate at 15 µg

      Secondary
      Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution

      Predicted band size: 58 kDa

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ICAM1 antibody [EPR4776] (ab109361)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ICAM1 antibody [EPR4776] (ab109361)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human lung carcinoma tissue sections labeling ICAM1 with Purified ab109361 at 1/100 dilution (6.79 µg/mL). Heat mediated antigen retrieval using Bond™ Epitope Retrieval Solution 2 (pH 9.0). Rabbit specific IHC polymer detection kit HRP/DAB (ab209101) was used as the secondary antibody. Negative control: PBS instead of the primary antibody. Hematoxylin was used as a counterstain.
    • Flow Cytometry (Intracellular) - Anti-ICAM1 antibody [EPR4776] (ab109361)
      Flow Cytometry (Intracellular) - Anti-ICAM1 antibody [EPR4776] (ab109361)

      Intracellular Flow Cytometry analysis of Ramos (Human Burkitt's lymphoma B lymphocyte) cells labeling ICAM1 with Purified ab109361 at 1/70 dilution (10 µg/mL) (Red). Cells were fixed with 4% Paraformaldehyde and permeabilised with 90% Methanol. A Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) secondary antibody was used at 1/2000. Isotype control - Rabbit monoclonal IgG (Black). Unlabeled control - Cell without incubation with primary antibody and secondary antibody (Blue).

       

    • Immunoprecipitation - Anti-ICAM1 antibody [EPR4776] (ab109361)
      Immunoprecipitation - Anti-ICAM1 antibody [EPR4776] (ab109361)
      Purified ab109361 at 1/30 dilution (2µg) immunoprecipitating ICAM1 in Raji whole cell lysate.
      Lane 1 (input): Raji (Human Burkitt's lymphoma B lymphocyte) whole cell lysate 10µg
      Lane 2 (+): ab109361 + Raji whole cell lysate.
      Lane 3 (-): Rabbit monoclonal IgG (ab172730) instead of ab109361 in Raji whole cell lysate.
      VeriBlot for IP Detection Reagent (HRP) (ab131366) (1/1000 dilution) was used for Western blotting.
      Blocking Buffer and concentration: 5% NFDM/TBST.
      Diluting buffer and concentration: 5% NFDM/TBST.
      Observed band size: 89 kDa
    • Immunocytochemistry/ Immunofluorescence - Anti-ICAM1 antibody [EPR4776] (ab109361)
      Immunocytochemistry/ Immunofluorescence - Anti-ICAM1 antibody [EPR4776] (ab109361)
      Immunocytochemistry analysis of Raji (Human Burkitt's lymphoma B lymphocyte) cells labeling ICAM1 with Purified ab109361 at 1/100 dilution (6.79 µg/mL). Cells were fixed in 4% Paraformaldehyde and permeabilized with 0.1% tritonX-100. Cells were counterstained with Ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) 1/200 dilution (2.5 µg/mL). Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) was used as the secondary antibody at 1/1000 dilution (2 µg/mL). DAPI (blue) was used as nuclear counterstain. PBS instead of the primary antibody was used as the secondary antibody only control.
    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ICAM1 antibody [EPR4776] (ab109361)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ICAM1 antibody [EPR4776] (ab109361)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human tonsil tissue sections labeling ICAM1 with Purified ab109361 at 1/100 dilution (6.79 µg/mL). Heat mediated antigen retrieval using Bond™ Epitope Retrieval Solution 2 (pH 9.0). Rabbit specific IHC polymer detection kit HRP/DAB (ab209101) was used as the secondary antibody. Negative control: PBS instead of the primary antibody. Hematoxylin was used as a counterstain.
    • Anti-ICAM1 antibody [EPR4776] (ab109361)
      Anti-ICAM1 antibody [EPR4776] (ab109361)

    Protocols

    • Flow cytometry protocols
    • Immunoprecipitation protocols
    • Immunohistochemistry protocols
    • Western blot protocols

    Click here to view the general protocols

    Datasheets and documents

    • SDS download

    • Datasheet download

      Download

    References (14)

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

    ab109361 has been referenced in 14 publications.

    • Li Q  et al. Single-cell transcriptome profiling reveals vascular endothelial cell heterogeneity in human skin. Theranostics 11:6461-6476 (2021). PubMed: 33995668
    • Yao MY  et al. Long non-coding RNA MALAT1 exacerbates acute respiratory distress syndrome by upregulating ICAM-1 expression via microRNA-150-5p downregulation. Aging (Albany NY) 12:6570-6585 (2020). PubMed: 32315984
    • Yang W  et al. Potential role of lncRNA HULC/miR-128-3p/RAC1 axis in the inflammatory response during LPS-induced sepsis in HMEC-1 cells. Mol Med Rep 22:5095-5104 (2020). PubMed: 33174038
    • Qiu D  et al. Hyperglycemia Decreases Epithelial Cell Proliferation and Attenuates Neutrophil Activity by Reducing ICAM-1 and LFA-1 Expression Levels. Front Genet 11:616988 (2020). PubMed: 33414814
    • Shishkova D  et al. Calciprotein Particles Cause Endothelial Dysfunction under Flow. Int J Mol Sci 21:N/A (2020). PubMed: 33233811
    View all Publications for this product

    Customer reviews and Q&As

    Show All Reviews Q&A
    Submit a review Submit a question

    1-4 of 4 Abreviews or Q&A

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

    Excellent
    Abreviews
    Abreviews
    abreview image
    Application
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
    Sample
    Human Tissue sections (Frontal Cortex (Brain))
    Antigen retrieval step
    Heat mediated - Buffer/Enzyme Used: Tris-EDTA/AR9
    Permeabilization
    Yes - 0.1% TritonX-100
    Specification
    Frontal Cortex (Brain)
    Blocking step
    Donkey Serum as blocking agent for 30 minute(s) · Concentration: 3% · Temperature: 25°C
    Fixative
    Paraformaldehyde
    Read More
    The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

    Abcam user community

    Verified customer

    Submitted Mar 18 2020

    Question

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

    Read More

    Abcam community

    Verified customer

    Asked on Jul 30 2012

    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.

    Read More

    Abcam Scientific Support

    Answered on Jul 30 2012

    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

    Abcam community

    Verified customer

    Asked on Jun 05 2012

    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

    Abcam Scientific Support

    Answered on Jun 05 2012

    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

    Abcam community

    Verified customer

    Asked on Jun 05 2012

    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

    Abcam Scientific Support

    Answered on Jun 05 2012

    Please note: All products are "FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC PROCEDURES"
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