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    icam1-antibody-ep1442y-low-endotoxin-azide-free-ab221777.pdf

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Immunology Cell Type Markers CD Adhesion
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RecombinantRabMAb

Recombinant Anti-ICAM1 antibody [EP1442Y] - Low endotoxin, Azide free (ab221777)

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  • Certificate of Compliance
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Western blot - Anti-ICAM1 antibody [EP1442Y] - Low endotoxin, Azide free (ab221777)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ICAM1 antibody [EP1442Y] - Low endotoxin, Azide free (ab221777)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ICAM1 antibody [EP1442Y] - Low endotoxin, Azide free (ab221777)
  • Western blot - Anti-ICAM1 antibody [EP1442Y] - Low endotoxin, Azide free (ab221777)
  • Western blot - Anti-ICAM1 antibody [EP1442Y] - Low endotoxin, Azide free (ab221777)
  • Anti-ICAM1 antibody [EP1442Y] - Low endotoxin, Azide free (ab221777)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EP1442Y] to ICAM1 - Low endotoxin, Azide free
  • Suitable for: WB, IHC-P
  • Reacts with: Human

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Overview

  • Product name

    Anti-ICAM1 antibody [EP1442Y] - Low endotoxin, Azide free
    See all ICAM1 primary antibodies
  • Description

    Rabbit monoclonal [EP1442Y] to ICAM1 - Low endotoxin, Azide free
  • Host species

    Rabbit
  • Tested applications

    Suitable for: WB, IHC-Pmore details
  • Species reactivity

    Reacts with: Human
  • Immunogen

    Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.

  • Positive control

    • WB: Huvec, Ramos and Raji whole cell lysate; IHC: Human kidney and tonsil tissue.
  • General notes

    ab221777 is the carrier-free version of ab53013.

    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 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.

    Our Low endotoxin, azide-free formats have low endotoxin level (≤ 1 EU/ml, determined by the LAL assay) and are free from azide, to achieve consistent experimental results in functional assays.

    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: 7.20
    Constituent: PBS
  • Carrier free

    Yes
  • Concentration information loading...
  • Purity

    Protein A purified
  • Clonality

    Monoclonal
  • Clone number

    EP1442Y
  • 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 [EP1442Y] - BSA and Azide free (ab271852)
    • Anti-ICAM1 antibody [EP1442Y] (ab53013)
  • Compatible Secondaries

    • Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077)
    • Goat Anti-Rabbit IgG H&L (HRP) (ab205718)
  • Conjugation kits

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

    • Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730)
  • KO cell lines

    • Human ICAM1 knockout HeLa cell line (ab261742)
  • KO cell lysates

    • Human ICAM1 knockout HeLa cell lysate (ab256947)
  • KO cell pellets

    • Human ICAM1 knockout HeLa cell pellet (ab278920)
  • Recombinant Protein

    • Recombinant Human ICAM1 protein (ab151393)

Applications

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab221777 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. Detects a band of approximately 89 kDa (predicted molecular weight: 58 kDa).
IHC-P
Use at an assay dependent concentration. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Notes
WB
Use at an assay dependent concentration. Detects a band of approximately 89 kDa (predicted molecular weight: 58 kDa).
IHC-P
Use at an assay dependent concentration. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

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 [EP1442Y] - Low endotoxin, Azide free (ab221777)
      Western blot - Anti-ICAM1 antibody [EP1442Y] - Low endotoxin, Azide free (ab221777)
      All lanes : Anti-ICAM1 antibody [EP1442Y] (ab53013) 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 [EP1442Y] 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, ab53013 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 (not observed by this antibody) 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.

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ICAM1 antibody [EP1442Y] - Low endotoxin, Azide free (ab221777)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ICAM1 antibody [EP1442Y] - Low endotoxin, Azide free (ab221777)
      This data was developed using ab53013, 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 ab53013 at 1/500 dilution (0.22 µg/mL). Heat mediated antigen retrieval using Bond™ Epitope Retrieval Solution 2 (pH 9.0). The immunostaining was performed on a Leica Biosystems BOND® RX instrument. 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.
    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ICAM1 antibody [EP1442Y] - Low endotoxin, Azide free (ab221777)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ICAM1 antibody [EP1442Y] - Low endotoxin, Azide free (ab221777)
      This data was developed using ab53013, the same antibody clone in a different buffer formulation.
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of Human kidney tissue sections labeling ICAM1 with purified ab53013 at 1/500 dilution (0.22 µg/mL). Heat mediated antigen retrieval using Bond™ Epitope Retrieval Solution 2 (pH 9.0). The immunostaining was performed on a Leica Biosystems BOND® RX instrument. 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.
    • Western blot - Anti-ICAM1 antibody [EP1442Y] - Low endotoxin, Azide free (ab221777)
      Western blot - Anti-ICAM1 antibody [EP1442Y] - Low endotoxin, Azide free (ab221777)
      All lanes : Anti-ICAM1 antibody [EP1442Y] - BSA and Azide free (ab271852) at 1/1000 dilution

      Lane 1 : Untreated HUVEC (Human umbilical vein endothelial cell) whole cell lysate
      Lane 2 : HUVEC (Human umbilical vein endothelial cell) treated with 50ng/ml TNF-a for 24 hours whole cell lysate
      Lane 3 : Raji (Human Burkitt's lymphoma B lymphocyte) whole cell lysate

      Lysates/proteins at 20 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



      The molecular mass observed is consistent with what has been described in the literatures (PMID: 29777158, 30082828, 31244919).

    • Western blot - Anti-ICAM1 antibody [EP1442Y] - Low endotoxin, Azide free (ab221777)
      Western blot - Anti-ICAM1 antibody [EP1442Y] - Low endotoxin, Azide free (ab221777)
      All lanes : Anti-ICAM1 antibody [EP1442Y] (ab53013) at 1/1000 dilution

      Lane 1 : Wild-type HeLa cell lysate
      Lane 2 : ICAM1 knockout HeLa cell lysate
      Lane 3 : Raji cell lysate
      Lane 4 : Ramos cell lysate

      Lysates/proteins at 20 µg per lane.

      Secondary
      All lanes : Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) at 1/10000 dilution

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



      This data was developed using ab53013, the same antibody clone in a different buffer formulation.

      Lanes 1-4: Merged signal (red and green). Green - ab53013 observed at 90 kDa. Red - loading control ab8245 observed at 36 kDa.

       ab53013 Anti-ICAM1 antibody [EP1442Y]  was shown to specifically react with ICAM1 in wild-type HeLa cells. Loss of signal was observed when knockout cell line ab261742 (knockout cell lysate ab256947) was used. Wild-type and ICAM1 knockout samples were subjected to SDS-PAGE. ab53013 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated overnight at 4°C at 1 in 1000 dilution and 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.

    • Anti-ICAM1 antibody [EP1442Y] - Low endotoxin, Azide free (ab221777)
      Anti-ICAM1 antibody [EP1442Y] - Low endotoxin, Azide free (ab221777)

    Protocols

    • Flow cytometry protocols
    • Immunohistochemistry protocols
    • Immunocytochemistry & immunofluorescence protocols
    • Western blot protocols

    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 (14)

    Publishing research using ab221777? Please let us know so that we can cite the reference in this datasheet.

    ab221777 has been referenced in 14 publications.

    • Shao X  et al. Astragaloside IV alleviates atherosclerosis through targeting circ_0000231/miR-135a-5p/CLIC4 axis in AS cell model in vitro. Mol Cell Biochem 476:1783-1795 (2021). PubMed: 33439448
    • Wei Y  et al. MicroRNA-19b-3p suppresses gastric cancer development by negatively regulating neuropilin-1. Cancer Cell Int 20:193 (2020). PubMed: 32508529
    • Shi H  et al. Long non-coding RNA expression profile in minor salivary gland of primary Sjögren's syndrome. Arthritis Res Ther 18:109 (2016). IHC ; Human . PubMed: 27188286
    • Broniarek I  et al. The effect of chronic exposure to high palmitic acid concentrations on the aerobic metabolism of human endothelial EA.hy926 cells. Pflugers Arch 468:1541-54 (2016). WB ; Human . PubMed: 27417103
    • Pannier D  et al. High expression levels of egfl7 correlate with low endothelial cell activation in peritumoral vessels of human breast cancer. Oncol Lett 12:1422-1428 (2016). IHC-P ; Human . PubMed: 27446447
    View all Publications for this product

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