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Human ICAM-1/CD54 Antibody

Catalog #: AF720
12 Citations 1 Review Datasheet / COA / SDS
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AF720
AF720-SP
Detection of Human ICAM‑1/CD54 by Western Blot.
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Product Details
Citations (12)
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Reviews (1)
Summary Data Examples Preparation and Storage Reconstitution Calculator Background Product Datasheets Related Research Areas

Human ICAM-1/CD54 Antibody Summary

Species Reactivity
Human
Specificity
Selected for its ability to block the adhesion of HSB‑2 cells to immobilized recombinant human (rh) ICAM‑1. In direct ELISAs, approximately 5% cross-reactivity with rhICAM-3 is observed and less than 2% cross-reactivity with rhICAM-2, rhICAM-4, rhICAM-5, recombinant rat ICAM‑1, recombinant mouse ICAM‑1, and rhVCAM-1 is observed.
Source
Polyclonal Sheep IgG
Purification
Antigen Affinity-purified
Immunogen
Chinese hamster ovary cell line CHO-derived recombinant human ICAM-1
Extracellular domain
Formulation
Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. *Small pack size (SP) is supplied either lyophilized or as a 0.2 µm filtered solution in PBS.
Endotoxin Level
<0.20 EU per 1 μg of the antibody by the LAL method.
Label
Unconjugated

Applications

Recommended Concentration
Sample
Western Blot
0.2 µg/mL
See below
Adhesion Blockade
The adhesion of HSB2 human peripheral blood acute lymphoblastic leukemia cells (5 x 104 cells/well) to immobilized recombinant human ICAM-1/Fc Chimera (12.5 μg/mL, 100 µL/well) was maximally inhibited (80‑100%) by 25 μg/mL of the antibody.
 
Immunocytochemistry
1-15 µg/mL
See below
Knockout Validated
ICAM‑1/CD54 is specifically detected in the parental Ramos human Burkitt's lymphoma cell line but is not detectable in knockout Ramos human Burkitt's lymphoma cell line.
 

Please Note: Optimal dilutions should be determined by each laboratory for each application. General Protocols are available in the Technical Information section on our website.

Scientific Data

Western Blot Detection of Human ICAM-1/CD54 antibody by Western Blot. View Larger

Detection of Human ICAM‑1/CD54 by Western Blot. Western blot shows lysates of HUVEC human umbilical vein endothelial cells and Ramos human Burkitt's lymphoma cell line untreated (-) or treated (+) with 10 ng/mL Recombinant Human TNF-a (Catalog # 210-TA) for 24 hours. PVDF membrane was probed with 0.2 µg/mL of Sheep Anti-Human ICAM-1/CD54 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF720) followed by HRP-conjugated Anti-Sheep IgG Secondary Antibody (Catalog # HAF016). A specific band was detected for ICAM-1/CD54 at approximately 90 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.

Immunocytochemistry ICAM-1/CD54 antibody in A431 Human Cell Line by Immunocytochemistry (ICC). View Larger

ICAM‑1/CD54 in A431 Human Cell Line. ICAM-1/CD54 was detected in immersion fixed A431 human epithelial carcinoma cell line (left panel; positive stain) and A549 human lung carcinoma cell line (right panel; negative stain) using Sheep Anti-Human ICAM-1/CD54 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF720) at 1.7 µg/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Anti-Sheep IgG Secondary Antibody (red; Catalog # NL010) and counterstained with DAPI (blue). Specific staining was localized to plasma membrane. View our protocol for Fluorescent ICC Staining of Cells on Coverslips.

Knockout Validated View Larger

Western Blot Shows Human ICAM‑1/CD54 Specificity by Using Knockout Cell Line. Western blot shows lysates of Ramos human Burkitt's lymphoma cell line and human ICAM-1/CD54 Ramos human Burkitt's lymphoma cell line (KO). PVDF membrane was probed with 0.2 µg/mL of Sheep Anti-Human ICAM‑1/CD54 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF720) followed by HRP-conjugated Anti-Sheep IgG Secondary Antibody (HAF016). A specific band was detected for ICAM‑1/CD54 at approximately 90 kDa (as indicated) in the parental Ramos human Burkitt's lymphoma cell line, but is not detectable in knockout Ramos human Burkitt's lymphoma cell line. GAPDH (Catalog # AF5718) is shown as a loading control. This experiment was conducted under reducing conditions and using Western Blot Buffer Group 1.

Reconstitution Calculator

Reconstitution Calculator

The reconstitution calculator allows you to quickly calculate the volume of a reagent to reconstitute your vial. Simply enter the mass of reagent and the target concentration and the calculator will determine the rest.

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Preparation and Storage

Reconstitution
Reconstitute at 0.2 mg/mL in sterile PBS.
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Shipping
Lyophilized product is shipped at ambient temperature. Liquid small pack size (-SP) is shipped with polar packs. Upon receipt, store immediately at the temperature recommended below.
Stability & Storage
Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
  • 12 months from date of receipt, -20 to -70 °C as supplied.
  • 1 month, 2 to 8 °C under sterile conditions after reconstitution.
  • 6 months, -20 to -70 °C under sterile conditions after reconstitution.

Background: ICAM-1/CD54

Intercellular Adhesion Molecule-1 (ICAM-1) binds the leukocyte integrins LFA-1 and Mac-1. ICAM-1 expression is weak on leukocytes, epithelial and resting endothelial cells, as well as some other cell types, but expression can be stimulated by IFN-gamma, TNF-alpha, IL-1 beta and LPS. Soluble ICAM-1 is found in a biologically active form in serum, probably as a result of proteolytic cleavage from the cell surface, and is elevated in patients with various inflammatory syndromes such as septic shock, LAD, cancer and transplantation.

Long Name
Intercellular Adhesion Molecule 1
Entrez Gene IDs
3383 (Human); 15894 (Mouse); 25464 (Rat)
Alternate Names
BB2; CD54 antigen; CD54; cell surface glycoprotein P3.58; human rhinovirus receptor; ICAM1; ICAM-1; intercellular adhesion molecule 1 (CD54), human rhinovirus receptor; intercellular adhesion molecule 1; Major group rhinovirus receptor; P3.58

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Citations for Human ICAM-1/CD54 Antibody

R&D Systems personnel manually curate a database that contains references using R&D Systems products. The data collected includes not only links to publications in PubMed, but also provides information about sample types, species, and experimental conditions.

12 Citations: Showing 1 - 10
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  1. Ras-mediated modulation of pyruvate dehydrogenase activity regulates mitochondrial reserve capacity and contributes to glioblastoma tumorigenesis
    Authors: Antony Prabhu, Bhaswati Sarcar, C. Ryan Miller, Sung-Hak Kim, Ichiro Nakano, Peter Forsyth et al.
    Neuro-Oncology
  2. Vascular and extravascular immunoreactivity for intercellular adhesion molecule 1 in the orbitofrontal cortex of subjects with major depression: Age-dependent changes
    Authors: Jose Javier Miguel-Hidalgo, James C. Overholser, George J. Jurjus, Herbert Y. Meltzer, Lesa Dieter, Lisa Konick et al.
    Journal of Affective Disorders
  3. T cell‐dependent bispecific antibodies alter organ‐specific endothelial cell–T cell interaction
    Authors: Patricia Himmels, Thi Thu Thao Nguyen, Maresa Caunt Mitzner, Alfonso Arrazate, Stacey Yeung, Jeremy Burton et al.
    EMBO reports
  4. ICAM1 initiates CTC cluster formation and trans-endothelial migration in lung metastasis of breast cancer
    Authors: R Taftaf, X Liu, S Singh, Y Jia, NK Dashzeveg, AD Hoffmann, L El-Shennaw, EK Ramos, V Adorno-Cru, EJ Schuster, D Scholten, D Patel, Y Zhang, AA Davis, C Reduzzi, Y Cao, P D'Amico, Y Shen, M Cristofani, WA Muller, V Varadan, H Liu
    Nature Communications, 2021-08-11;12(1):4867.
    Species: Human
    Sample Types: Whole Cells
    Applications: Neutralization
  5. Multiplexed, targeted profiling of single-cell proteomes and transcriptomes in a single reaction
    Genome Biol, 2016-09-19;17(1):188.
    Species: Human
    Sample Types: Whole Cells
    Applications: ICC
  6. High molecular weight hyaluronic acid inhibits IL-6-induced MMP production from human chondrocytes by up-regulating the ERK inhibitor, MKP-1.
    Authors: Hashizume M, Mihara M
    Biochem. Biophys. Res. Commun., 2010-11-06;403(2):184-9.
    Species: Human
    Sample Types: Whole Cells
    Applications: Neutralization
  7. C-reactive protein (CRP) induces chemokine secretion via CD11b/ICAM-1 interaction in human adherent monocytes.
    Authors: Montecucco F, Steffens S, Burger F, Pelli G, Monaco C, Mach F
    J. Leukoc. Biol., 2008-06-30;84(4):1109-19.
    Species: Human
    Sample Types: Whole Cells
    Applications: Neutralization
  8. Endothelial cells of human colorectal cancer and healthy colon reveal phenotypic differences in culture.
    Authors: Schellerer VS, Croner RS, Weinlander K, Hohenberger W, Sturzl M, Naschberger E
    Lab. Invest., 2007-09-17;87(11):1159-70.
    Species: Human
    Sample Types: Whole Cells
    Applications: ICC
  9. The role of vascular cell adhesion molecule 1/ very late activation antigen 4 in endothelial progenitor cell recruitment to rheumatoid arthritis synovium.
    Authors: Silverman MD, Haas CS, Rad AM, Arbab AS, Koch AE
    Arthritis Rheum., 2007-06-01;56(6):1817-26.
    Species: Human
    Sample Types: Cell Lysates, Whole Cells
    Applications: Neutralization, Western Blot
  10. Shear flow-dependent integration of apical and subendothelial chemokines in T-cell transmigration: implications for locomotion and the multistep paradigm.
    Authors: Schreiber TH, Shinder V, Cain DW, Alon R, Sackstein R
    Blood, 2006-10-12;109(4):1381-6.
    Species: Human
    Sample Types: Whole Cells
    Applications: Neutralization
  11. Cellular entry and uncoating of naked and quasi-enveloped human hepatoviruses
    Authors: Rivera-Serrano EE, Gonzalez-Lopez O, Das A, Lemon SM.
    Elife
  12. Endothelial P2Y2 receptor regulates LPS-induced neutrophil transendothelial migration in vitro
    Authors: Filip Kukulski, Fethia Ben Ben Yebdri, Fariborz Bahrami, Michel Fausther, Alain Tremblay, Jean Sévigny
    Molecular Immunology

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Human ICAM‑1/CD54 Antibody
By Anonymous on 03/23/2018
Application: WB Sample Tested: Purified protein Species: Mouse