Key Product Details

Species Reactivity

Validated:

Human

Cited:

Human, Mouse, Rat, Insect - Aedes albopictus (Asian tiger mosquito)

Applications

Validated:

Western Blot, Adhesion Blockade, Flow Cytometry, Immunocytochemistry, Immunoprecipitation

Cited:

Immunohistochemistry, Immunohistochemistry-Frozen, Western Blot, Neutralization, Flow Cytometry, Immunofluorescence, Immunocytochemistry, Immunoprecipitation, Bead-based Bioassay, Bioassay, Cell-based ELISA, ELISA Capture, ELISA Development, ELISA Development (Capture), Functional Assay, Inhibition

Label

Unconjugated

Antibody Source

Monoclonal Mouse IgG1 Clone # BBIG-I1 (11C81)
View all ICAM-1/CD54 Primary Antibodies »

Product Specifications

Immunogen

Activated HUVEC human umbilical vein endothelial cells

Specificity

This antibody was selected for its ability to stain human ICAM-1/CD54 transfected COS cells. It does not stain COS cells transfected with human E-Selectin, VCAM-1, or PECAM-1.

Clonality

Monoclonal

Host

Mouse

Isotype

IgG1

Endotoxin Level

<0.10 EU per 1 μg of the antibody by the LAL method.

Scientific Data Images for Human ICAM‑1/CD54 Antibody

Detection of Human ICAM‑1/CD54 by Western Blot.

Western blot shows lysates of C32. PVDF membrane was probed with 2 µg/mL of Mouse Anti-Human ICAM‑1/CD54 Monoclonal Antibody (Catalog # BBA3) followed by HRP-conjugated Anti-Mouse IgG Secondary Antibody (Catalog # HAF018). A specific band was detected for ICAM‑1/CD54 at approximately 85 kDa (as indicated). This experiment was conducted under non-reducing conditions and using Western Blot Buffer Group 1.

ICAM‑1/CD54 in C32 Human Cell Line.

ICAM‑1/CD54 was detected in live unfixed C32 human melanoma cell line using Mouse Anti-Human ICAM‑1/CD54 Monoclonal Antibody (Catalog # BBA3) at 5 µg/mL for 1 hour at 2° - 8° C. Cells were then fixed with 4% formaldehyde followed by incubation with the NorthernLights™ 557-conjugated Anti-Mouse IgG Secondary Antibody (red; NL007) and counterstained with DAPI (blue). Specific staining was localized to cell surface and cytoplasm.
Detection of ICAM-1/CD54 by Immunohistochemistry

Detection of ICAM-1/CD54 by Immunohistochemistry

SERS-BFNP molecular imaging of atherosclerotic coronary arteries. A single human coronary artery was isolated from the heart of a patient undergoing heart transplantation surgery. The lumen of the artery segment was then injected with a mixture of anti-ICAM-1, anti-VCAM-1, anti-P-selectin, and isotype control BFNP, sutured closed, and incubated at 37 °C/5% CO2 for 12 h. Sutures were then removed and the artery segment was thoroughly washed prior to SERS spectroscopy and subsequent analysis of morphology, expression of adhesion molecules and SERS mapping. (B) Immunofluorescence staining for CD31, and expression of (C) ICAM-1, VCAM-1 and P-selectin are shown in red. Nuclei were counterstained using Hoechst 33342 (blue). Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/30613292), licensed under a CC-BY license. Not internally tested by R&D Systems.

Detection of ICAM‑1/CD54 in Human PBMCs by Flow Cytometry.

Human peripheral blood mononuclear cells (PBMCs) were stained with (A) Mouse Anti-Human ICAM-1/CD54 Monoclonal Antibody (Catalog # BBA3) or (B) Mouse IgG1 Isotype Control (MAB002) followed by anti-Mouse IgG PE-conjugated Secondary Antibody (F0102B) and Mouse Anti-Human CD14 APC-conjugated Monoclonal Antibody (FAB3832A). Staining was performed using our Staining Membrane-associated Proteins protocol.
Detection of ICAM-1/CD54 by Immunohistochemistry

Detection of ICAM-1/CD54 by Immunohistochemistry

In vivo SERS-BFNP molecular imaging of adhesion molecules. Following engraftment of human adipose, HANSG mice were allowed to recover for 3 weeks. Mice were then injected intravenously with 5 μg of human recombinant TNF-alpha 4 h prior to receiving an intravenous injection of BFNP. (A) Following SERS-BFNP molecular imaging, adipose grafts were excised and immunofluorescently stained for human (red) and murine (green) CD31. (B) Isotype control, ICAM-1, VCAM-1, and P-selectin staining are also shown in white counterstained with human CD31 (red). Nuclei were counterstained using Hoechst 33342 (blue). (C) To conduct SERS-BFNP molecular imaging, HANSG mice were anaesthetized and their adipose grafts non-invasively analyzed in vivo using SERS spectroscopy. (D) SERS spectra were acquired from mice that received a mixture of isotype-PPY, -BPE and -PYOT (blue spectra), or anti-P-selectin-PPY, anti-ICAM-1-BPE, anti-VCAM-1-PYOT BFNP (red spectra). The spectra shown are from 5 isotype vs. 5 targeted mice, with each spectrum acquired from a different mouse. (E) In addition to immunofluorescence microscopy, excised adipose grafts were analyzed using SERS microscopy. Detection of BFNP from sections of adipose tissue isolated from HANSG mice that received anti-ICAM-1 (purple), anti-VCAM-1 (red), and anti-P-selectin (blue) (upper panels) or Isotype-BPE (purple), Isotype-PYOT (red), and Isotype-PPY (blue) (lower panels) are shown superimposed on darkfield tissue images alongside a magnified image of Raman maps from the scanned areas (black boxes). The colored circles in the Raman map ((E) upper panel) correlate to the acquired spectra shown in (F) above their respective reference spectra. The optical image in (E) is a darkfield image. Scale bars: (A) = 1000 μm; (B) = 100 μm; (E) = 20 μm. Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/30613292), licensed under a CC-BY license. Not internally tested by R&D Systems.
Detection of ICAM-1/CD54 by Immunocytochemistry/ Immunofluorescence

Detection of ICAM-1/CD54 by Immunocytochemistry/ Immunofluorescence

Following stimulation, coronary artery endothelial cells (CAEC) express adhesion molecules detectable via immuno-SERS imaging in single and multiplex formats. (A) Fluorescence images of immunohistochemical staining of ICAM-1, VCAM-1 and P-selectin on CAEC in unstimulated and 10 ng/mL TNF-alpha -stimulated conditions. Isotype control, ICAM-1, VCAM-1 and P-selectin staining shown in green; nuclei were counterstained using Hoechst 33342 (blue). (B) CAEC were stimulated with 10 ng/mL TNF-alpha for 24 h, fixed in acetone, and incubated with isotype control, anti-ICAM-1, anti-VCAM-1 or anti-P-selectin BFNP or (C) with all BFNP simultaneously before being subjected to SERS mapping. (D) Representative spectra from anti-ICAM-1 (purple), anti-VCAM-1 (red) and anti-P-selectin (blue) BFNP acquired from the color-matched circles in (C) are shown above their respective reference spectra. Optical images in (B-C) are darkfield images. Scale bars = 20 μm. Results are representative of 3 independent experiments. Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/30613292), licensed under a CC-BY license. Not internally tested by R&D Systems.
Detection of ICAM-1/CD54 by Western Blot

Detection of ICAM-1/CD54 by Western Blot

Methotrexate (MTX) and Ad repress TNF-alpha -induced pro-inflammatory genes without affecting primary MIR181A-1 and MIR181B-1 expression.(A) Western blot analysis of VCAM-1, ICAM-1, and E-Selectin in HUVECs treated with or without MTX (10 µM) or Ad (50 µM), after stimulation of TNF-alpha (10 ng/ml) for 8 hr. Quantification of n = 3 independent experiments. (B) Real-time qPCR analysis of VCAM-1, ICAM-1, and E-Selectin in HUVECs treated with or without MTX (10 µM) or Ad (50 µM), after treatment with TNF-alpha (10 ng/ml) for 4 hr. Real-time qPCR analysis of (C) primary transcript of MIR181A1 or (D) primary transcript of MIR181B-1 in HUVECs treated with or without MTX (10 µM) or Ad (50 µM), after treatment with TNF-alpha (10 ng/ml) for 4 hr. (A–D), n = 3–6. *p<0.05; **p<0.01; ***p<0.001; ***p<0.0001. All values represent mean ± SEM. Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/33416495), licensed under a CC-BY license. Not internally tested by R&D Systems.
Detection of ICAM-1/CD54 by Western Blot

Detection of ICAM-1/CD54 by Western Blot

Induction of MIR181B expression by methotrexate (MTX) or adenosine is adenosine receptor A3 (ADORA3) dependent.(A) Knockdown for adenosine receptors A1, A2A, A2B, and A3 in HUVECs was performed to analyze MIR181B expression. three biological replicates. One-way ANOVA. (B) MIR181B expression in HUVECs transfected with Ctl-siRNA or ADORA3 siRNA after treatment with MTX (10 µM) or Ad (50 µM) or (C) treatment with TNF-alpha (10 ng/ml) alone or in combination MTX (10 µM) or Ad (50 µM). Three biological replicates. One-way ANOVA and Unpaired two-tailed Student t test. (D) Western blot analyses of VCAM-1, ICAM-1, and E-Selectin expression in HUVECs transfected with Ctl-siRNA or ADORA3 siRNA in the presence of TNF-alpha (10 ng/ml) in combination with either MTX (10 µM) or Ad (50 µM). Three biological replicates. Unpaired two-tailed Student t test. (E) in the presence of miRNA negative control (NS-m) or MIR181B mimics (181b-m) stimulated with TNF-alpha (10 ng/ml) or in combination with MTX (10 µM) or Ad (50 µM). Please see Figure 2—source data 1. Three biological replicates. Unpaired two-tailed Student t test. *p<0.05; **p<0.01; ***p<0.001; ****p<0.0001. n.s. indicated non significance. All values represent mean ± SEM.Figure 2—source data 1.Knockdown efficiency for adenosine receptor siRNAs.mRNA, and protein expression analysis for (A, B) ADORA3 siRNA, (C, D) ADORA1 siRNA, (E, F) ADORA2A siRNA and (G, H) ADORA2B siRNA compared to control siRNA in HUVECs transfected for 36 hr. (A–H), n = 3. ***p<0.001. All values represent mean ± SEM. Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/33416495), licensed under a CC-BY license. Not internally tested by R&D Systems.

Applications for Human ICAM‑1/CD54 Antibody

Application
Recommended Usage

Adhesion Blockade

The adhesion of HSB2 human peripheral blood acute lymphoblastic leukemia cells (5 x 104 cells/well) to immobilized Recombinant Human ICAM-1/CD54 Fc Chimera (Catalog # 720-IC,12.5 µg/mL, 100 µL/well) was maximally inhibited (80-100%) by 10 µg/mL of the antibody.

Flow Cytometry

0.25 µg/106 cells
Sample: Human PBMC

Immunocytochemistry

5-25 µg/mL
Sample: Live, unfixed C32 human melanoma cell line

Immunoprecipitation

Davies, M.E., et al. (1991) Connect. Tissue Res. 26:207.

Western Blot

2 µg/mL
Sample: C32 human melanoma cell line

Reviewed Applications

Read 7 reviews rated 4.9 using BBA3 in the following applications:

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Flow Cytometry

Formulation, Preparation, and Storage

Purification

Protein A or G purified from hybridoma culture supernatant

Reconstitution

Sterile PBS to a final concentration of 0.5 mg/mL.

Formulation

Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose.

Shipping

The product is shipped at ambient temperature. Upon receipt, store it 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.

Calculators

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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Background: ICAM-1/CD54

ICAM-1 is a member of the immunoglobulin superfamily whose expression is upregulated on leukocytes, epithelial cells and resting endothelial cells in response to inflammatory signals. ICAM-1 binds the leukocyte integrins LFA-1 and Mac-1.

Long Name

Intercellular Adhesion Molecule 1

Alternate Names

CD54, ICAM1

Entrez Gene IDs

3383 (Human); 15894 (Mouse); 25464 (Rat)

Gene Symbol

ICAM1

Additional ICAM-1/CD54 Products

Product Documents for Human ICAM‑1/CD54 Antibody

Certificate of Analysis

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Product Specific Notices for Human ICAM‑1/CD54 Antibody

For research use only

Citations for Human ICAM‑1/CD54 Antibody

Customer Reviews for Human ICAM‑1/CD54 Antibody (7)

4.9 out of 5
7 Customer Ratings
5 Stars
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Customer Images

Showing  1 - 5 of 7 reviews Showing All
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  • Human ICAM-1/CD54 Antibody
    Name: Anonymous
    Application: Immunocytochemistry/Immunofluorescence
    Sample Tested: HUVEC human umbilical vein endothelial cells
    Species: Human
    Verified Customer | Posted 07/10/2025
    Human ICAM‑1/CD54 Antibody BBA3
  • Human ICAM-1/CD54 Antibody
    Name: Anonymous
    Application: Immunocytochemistry/Immunofluorescence
    Sample Tested: Endothelial cells
    Species: Human
    Verified Customer | Posted 08/12/2021
    Human ICAM‑1/CD54 Antibody BBA3
  • Human ICAM-1/CD54 Antibody
    Name: Michelle Chen
    Application: Immunocytochemistry/Immunofluorescence
    Sample Tested: HUVEC human umbilical vein endothelial cells
    Species: Human
    Verified Customer | Posted 04/26/2017
    HUVECs were stimulated with LPS (10 ng/mL)
    Human ICAM‑1/CD54 Antibody BBA3
  • Name: Anonymous
    Application: Immunofluorescence
    Sample Tested: See PMID: 21277303
    Species: Human
    Verified Customer | Posted 01/20/2015
  • Name: Anonymous
    Application: Immunocytochemistry
    Sample Tested: See PMID: 23104645
    Species: Human
    Verified Customer | Posted 01/20/2015
  • Name: Anonymous
    Application: Immunofluorescence
    Sample Tested: See PMID: 21164106
    Species: Human
    Verified Customer | Posted 01/20/2015
  • Name: Anonymous
    Application: Immunofluorescence
    Sample Tested: See PMID: 22560286
    Species: Human
    Verified Customer | Posted 01/20/2015

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Showing  1 - 5 of 7 reviews Showing All

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