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    icam1-antibody-epr4776-bsa-and-azide-free-ab226059.pdf

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RecombinantRabMAb

Recombinant Anti-ICAM1 antibody [EPR4776] - BSA and Azide free (ab226059)

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Western blot - Anti-ICAM1 antibody [EPR4776] - BSA and Azide free (ab226059)
  • Western blot - Anti-ICAM1 antibody [EPR4776] - BSA and Azide free (ab226059)
  • Western blot - Anti-ICAM1 antibody [EPR4776] - BSA and Azide free (ab226059)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ICAM1 antibody [EPR4776] - BSA and Azide free (ab226059)
  • Flow Cytometry (Intracellular) - Anti-ICAM1 antibody [EPR4776] - BSA and Azide free (ab226059)
  • Immunoprecipitation - Anti-ICAM1 antibody [EPR4776] - BSA and Azide free (ab226059)
  • Immunocytochemistry/ Immunofluorescence - Anti-ICAM1 antibody [EPR4776] - BSA and Azide free (ab226059)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ICAM1 antibody [EPR4776] - BSA and Azide free (ab226059)
  • Anti-ICAM1 antibody [EPR4776] - BSA and Azide free (ab226059)

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 - BSA and Azide free
  • Suitable for: Flow Cyt (Intra), WB, IP, IHC-P, ICC/IF
  • Reacts with: Human

Conjugates logo Related conjugates and formulations

Alexa Fluor® 488 Alexa Fluor® 647 Biotin Unconjugated

You may also be interested in

Primary
Product image
Anti-ICAM1 antibody [EPR4776] (ab109361)
Knockout
Product image
Human ICAM1 knockout HeLa cell line (ab261742)

View more associated products

Overview

  • Product name

    Anti-ICAM1 antibody [EPR4776] - BSA and Azide free
    See all ICAM1 primary antibodies
  • Description

    Rabbit monoclonal [EPR4776] to ICAM1 - BSA and Azide free
  • Host species

    Rabbit
  • Tested applications

    Suitable for: Flow Cyt (Intra), WB, IP, IHC-P, ICC/IFmore 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

    ab226059 is the carrier-free version of ab109361.

    Our carrier-free antibodies are typically supplied in a PBS-only formulation, purified and free of BSA, sodium azide and glycerol. The carrier-free buffer and high concentration allow for increased conjugation efficiency.

    This conjugation-ready format is designed for use with fluorochromes, metal isotopes, oligonucleotides, and enzymes, which makes them ideal for antibody labelling, functional and cell-based assays, flow-based assays (e.g. mass cytometry) and Multiplex Imaging applications.

    Use our conjugation kits for antibody conjugates that are ready-to-use in as little as 20 minutes with <1 minute hands-on-time and 100% antibody recovery: available for fluorescent dyes, HRP, biotin and gold.

    This product is compatible with the Maxpar® Antibody Labeling Kit from Fluidigm, without the need for antibody preparation. Maxpar® is a trademark of Fluidigm Canada Inc.

    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. Do Not Freeze.
  • Storage buffer

    pH: 7.2
    Constituent: PBS
  • Carrier free

    Yes
  • 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

    • Anti-ICAM1 antibody [EPR4776] (ab109361)
    • Alexa Fluor® 488 Anti-ICAM1 antibody [EPR4776] (ab214860)
    • Alexa Fluor® 647 Anti-ICAM1 antibody [EPR4776] (ab214944)
    • Biotin Anti-ICAM1 antibody [EPR4776] (ab217075)
  • Compatible Secondaries

    • Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077)
  • Conjugation kits

    • PE / R-Phycoerythrin Conjugation Kit - Lightning-Link® (ab102918)
    • APC Conjugation Kit - Lightning-Link® (ab201807)
  • Isotype control

    • Rabbit IgG, monoclonal [EPR25A] - Isotype Control - BSA and Azide Free (ab210849)
  • Positive Controls

    • Raji membrane extract lysate (ab30126)

Applications

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab226059 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)
Use at an assay dependent concentration.
WB
Use at an assay dependent concentration. Detects a band of approximately 89 kDa (predicted molecular weight: 58 kDa).
IP
Use at an assay dependent concentration.
IHC-P
Use at an assay dependent concentration. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

See IHC antigen retrieval protocols.

ICC/IF
Use at an assay dependent concentration.
Notes
Flow Cyt (Intra)
Use at an assay dependent concentration.
WB
Use at an assay dependent concentration. Detects a band of approximately 89 kDa (predicted molecular weight: 58 kDa).
IP
Use at an assay dependent concentration.
IHC-P
Use at an assay dependent concentration. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

See IHC antigen retrieval protocols.

ICC/IF
Use at an assay dependent concentration.

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] - BSA and Azide free (ab226059)
      Western blot - Anti-ICAM1 antibody [EPR4776] - BSA and Azide free (ab226059)
      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 knockout cell line ab261742 (knockout 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] - BSA and Azide free (ab226059)
      Western blot - Anti-ICAM1 antibody [EPR4776] - BSA and Azide free (ab226059)

      This data was developed using ab109361, the same antibody clone in a different buffer formulation.

    • Western blot - Anti-ICAM1 antibody [EPR4776] - BSA and Azide free (ab226059)
      Western blot - Anti-ICAM1 antibody [EPR4776] - BSA and Azide free (ab226059)
      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



      This data was developed using ab109361, the same antibody clone in a different buffer formulation.

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ICAM1 antibody [EPR4776] - BSA and Azide free (ab226059)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ICAM1 antibody [EPR4776] - BSA and Azide free (ab226059)
      This data was developed using ab109361, the same antibody clone in a different buffer formulation.
      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] - BSA and Azide free (ab226059)
      Flow Cytometry (Intracellular) - Anti-ICAM1 antibody [EPR4776] - BSA and Azide free (ab226059)

      This data was developed using ab109361, the same antibody clone in a different buffer formulation. 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] - BSA and Azide free (ab226059)
      Immunoprecipitation - Anti-ICAM1 antibody [EPR4776] - BSA and Azide free (ab226059)
      This data was developed using ab109361, the same antibody clone in a different buffer formulation.
      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] - BSA and Azide free (ab226059)
      Immunocytochemistry/ Immunofluorescence - Anti-ICAM1 antibody [EPR4776] - BSA and Azide free (ab226059)
      This data was developed using ab109361, the same antibody clone in a different buffer formulation.
      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] - BSA and Azide free (ab226059)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ICAM1 antibody [EPR4776] - BSA and Azide free (ab226059)
      This data was developed using ab109361, the same antibody clone in a different buffer formulation.
      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] - BSA and Azide free (ab226059)
      Anti-ICAM1 antibody [EPR4776] - BSA and Azide free (ab226059)

    Protocols

    To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.

    Click here to view the general protocols

    Datasheets and documents

    • Datasheet download

      Download

    Certificate of Compliance

    To download a Certificate of Compliance, please enter your Lot number below:

    References (0)

    Publishing research using ab226059? Please let us know so that we can cite the reference in this datasheet.

    ab226059 has not yet been referenced specifically in any publications.

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