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CD54 antibody | 15.2

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Anti Human CD54 Antibody, clone 15.2 thumbnail image 1 Anti Human CD54 Antibody, clone 15.2 thumbnail image 2 Anti Human CD54 Antibody, clone 15.2 thumbnail image 3 Anti Human CD54 Antibody, clone 15.2 thumbnail image 4 Anti Human CD54 Antibody, clone 15.2 thumbnail image 5 Anti Human CD54 Antibody, clone 15.2 thumbnail image 6 Anti Human CD54 Antibody, clone 15.2 thumbnail image 7 Anti Human CD54 Antibody, clone 15.2 thumbnail image 8 Anti Human CD54 Antibody, clone 15.2 thumbnail image 9 Anti Human CD54 Antibody, clone 15.2 thumbnail image 10
Anti Human CD54 Antibody, clone 15.2 gallery image 1
Figure A. Unstimulated cells stained with RPE conjugated Mouse anti Human CD31 (MCA1738PE) and Alexa Fluor A700 conjugated Mouse anti Human CD54 (MCA1615A700). Figure B. Cells stimulated with TNFα for 19 h then stained with RPE conjugated Mouse anti Human CD31 (MCA1738PE) and Alexa Fluor A700 conjugated Mouse anti Human CD54 (MCA1615A700). All experiments performed on HUVEC cells gated on live single cells in the presence of 10% human serum. Data acquired on the ZE5 Cell Analyzer
Anti Human CD54 Antibody, clone 15.2 gallery image 2
Figure A. FITC conjugated Mouse anti Human CD19 (MCA2495F) and Alexa Fluor 647 conjugated Mouse IgG1 isotype control (MCA928A647). Figure B. FITC conjugated Mouse anti Human CD19 (MCA2495F) and Alexa Fluor 647 conjugated Mouse anti Human CD54 (MCA1615A647). All experiments performed on red cell lysed human peripheral blood gated on lymphocytes in the presence of Human SeroBlock (BUF070A).
Anti Human CD54 Antibody, clone 15.2 gallery image 3
Figure A. FITC conjugated Mouse anti Human CD40 (MCA1590PE) and Alexa Fluor 647 conjugated Mouse IgG1 isotype control (MCA928A647). Figure B. FITC conjugated Mouse anti Human CD40 (MCA1590PE) and Alexa Fluor 647 conjugated Mouse anti Human CD54 (MCA1615A647). All experiments performed on red cell lysed human peripheral blood gated on lymphocytes in the presence of Human SeroBlock (BUF070A).
Anti Human CD54 Antibody, clone 15.2 gallery image 4
Published customer image:
Schematic representation of ligand binding sites on CD54 and epitope representation for Mouse anti Human CD54 antibody, clone 15.2 (MCA1615). Effect of blocking of ICAM-1: ligand interaction by Mouse anti Human CD54 antibody, clone 15.2 (MCA1615GA) on Ifn-γ production by NK cells in vitro.
Image caption:
Engagement of ICAM-1 with its cellular ligand but not with PfEMP1 is required for NK cell IFN-γ production. (A) Diagram of an ICAM-1 molecule showing schematic binding sites for LFA-1, Mac-1, CD11c/CD18 and PfEMP1. The epitope map of the anti-ICAM-1 mAb 15.2, My13 and RR1/1 is indicated. (B) Human PBMC were cultured with uninfected RBC (RBC, black bars) or with RBC infected with the 3D7 Pf strain (3D7, grey bars) in presence or absence of antibodies directed against NKG2D (isotype control), ICAM-1 or CD18. Three different clones of anti-ICAM-1 were used: 15.2 blocks the interaction of ICAM-1 with LFA-1 and with PfEMP1, RR1/1 blocks only the interaction with LFA-1 and My13 blocks only the interaction with PfEMP1. After 24 h of co-culture, NK cell activation was analyzed by flow cytometry by gating on CD3−CD56+ NK cells. The CD69 MFI staining on NK cells (left panel), the percentage of CD25+ NK cells (middle panel) and the percentage of IFN-γ+ NK cells (right panel) were determined for 24 donors (None, NKG2D and 15.2), 13 donors (CD18), 9 donors (My13) or 5 donors (RR1/1). Means ± SEM are represented. Statistical analyses were performed using the Mann Whitney test.

From: Baratin M, Roetynck S, Pouvelle B, Lemmers C, Viebig NK, Johansson S, et al. (2007)
Dissection of the Role of PfEMP1 and ICAM-1 in the Sensing of Plasmodium falciparum-Infected Erythrocytes by Natural Killer Cells.
PLoS ONE 2(2): e228.
This is from an open access article distributed under the terms of the Creative Commons Attribution License.
Anti Human CD54 Antibody, clone 15.2 gallery image 5
Published customer image:
Mouse anti Human CD54 antibody, clone 15.2 (MCA1615GA) used for the evaluation of ICAM-1 (CD54) expression on NK cells of differing origins by flow cytometry.
Image caption:
Primary resting human NK cells (CD56+CD3−) in total PBMC as well as human NK cell lines, NK92 and NKL were analyzed by flow cytometry to determine the surface expression of three host ligands for PfEMP1: CSA (left panels, dark line), CD36 (middle panels, dark line) and ICAM-1 (right panels, dark line). Stainings with isotype control for each antibody are represented by filled grey histograms. Stainings of primary resting human NK cells are representative of at least 3 donors.

From: Baratin M, Roetynck S, Pouvelle B, Lemmers C, Viebig NK, Johansson S, et al. (2007)
Dissection of the Role of PfEMP1 and ICAM-1 in the Sensing of Plasmodium falciparum-Infected Erythrocytes by Natural Killer Cells.
PLoS ONE 2(2): e228.
This is from an open access article distributed under the terms of the Creative Commons Attribution License.
Anti Human CD54 Antibody, clone 15.2 gallery image 6
Published customer image:
Mouse anti Human CD54 antibody, clone 15.2 (MCA1615GA) used to demonstrate co-localization of endogenous ICAM-1 with ICAM-1 GFP in human endothelial cells by immunofluorescence.
Image caption:
ICAM-1-GFP co-localizes with endogenous ICAM-1 and associates to filamin and actin upon clustering. (A) TNF-α-stimulated endothelium was transiently transfected with ICAM-1-GFP, fixed and stained for ICAM-1-GFP in green, endogenous ICAM-1 in red. Merge shows co-localization in yellow and F-actin is in grayscale. Images were taken from the apical surface (upper panel) and at the baso-lateral plane (lower panel). Bars, 50 μm. (B) Beads, coated with ICAM-1 Ab, were allowed to adhere for 30 minutes to TNF-α-stimulated HUVECs. ICAM-1 was stained using an ICAM-1 Ab that is directly labelled with ALEXA 647. The results show that ICAM-1-GFP (green) and endogenous ICAM-1 (red) co-localize (Arrowheads; Merge in yellow) at sites of bead adhesion. DIC image shows localization of the beads. Images were taken from the apical surface (upper panel) and at the baso-lateral plane (lower panel). Bars, 10 μm. (C) ICAM-1-GFP was transfected into HeLa cells and anti-ICAM-1-coated magnetic beads were allowed to adhere for 30 minutes, resulting in clustering of ICAM-1, after which the cells were lysed ands treated as described in the legend of figure 2C. The images on the left show that the magnetic beads efficiently precipitated ICAM-1-GFP and that ICAM-1 clustering resulted in the recruitment of actin and filamin A. Images on the right show the expression of indicated proteins in the cell lysates. (D) ICAM-1-GFP was transfected into HeLa cells and anti-ICAM-1-coated magnetic beads were allowed to adhere for 5 or 20 minutes, as indicated. ICAM-1 was precipitated at both time points, whereas actin and filamin A and B were associated to ICAM-1 only after 20 minutes of clustering. Images on the right show the expression of indicated proteins in the cell lysates.

From: van Buul JD, van Rijssel J, van Alphen FPJ, Hoogenboezem M, Tol S, Hoeben KA, et al. (2010)
Inside-Out Regulation of ICAM-1 Dynamics in TNF-α-Activated Endothelium.
PLoS ONE 5(6): e11336.
This is from an open access article distributed under the terms of the Creative Commons Attribution License.
Anti Human CD54 Antibody, clone 15.2 gallery image 7
FFPE human spleen stained with Mouse Anti-Human CD54 (MCA1615GA), biotinylated Anti-Mouse IgG and Streptavidin Alkaline Phosphatase (red). Nuclei were counterstained with hematoxylin.
Anti Human CD54 Antibody, clone 15.2 gallery image 8
Figure A. FITC conjugated Mouse anti Human CD19 (MCA1940F) and Alexa Fluor 647 conjugated Mouse IgG1 isotype control (MCA928A647). Figure B. FITC conjugated Mouse anti Human CD19 (MCA1940F) and Alexa Fluor 647 conjugated Mouse anti Human CD54 (MCA1615A647). All experiments performed on human blood gated on live single lymphocytes, in the presence of 10% human serum. Data acquired on the ZE5 Cell Analyzer.
Anti Human CD54 Antibody, clone 15.2 gallery image 9
Mouse anti Human CD54 (MCA1615GA) used to stain formalin fixed, paraffin embedded human spleen sections by immunohistochemistry, followed by biotinylated anti- mouse IgG and streptavidin alkaline phosphatase (red). Nuclei were counterstained with hematoxylin (blue).
Anti Human CD54 Antibody, clone 15.2 gallery image 10
Published customer image:
Mouse anti Human CD54 antibody, clone 15.2 (MCA1615) used to label cultivated HUVECs by immunofluorescence.
Image caption:
Endothelial CX3CL1 induces firm adhesion of CTLs to enhance transcellular diapedesis
(A–C) HUVECs were grown in μ-channels and activated with 10ng mL−1 for 20 h using TNFα. CTLs were flown over the activated HUVECs at physiological flow. HUVECs were incubated with 10μg mL−1 of blocking antibody for 60 min before the flow experiments. Blocking CX3CL1 using antibodies did not decrease transmigration efficiency (A) but resulted in a shift to more paracellular diapedesis events (B) and more distantly transmigrating CTL (C) (data are of CTL from 6 donors). Data are mean ±SD; ∗∗∗∗ p <0.0001. (D) HUVECs were cultured as described before and stimulated for 20 h with TNFα. Isolated CTLs (3 donors were analyzed) were added to the HUVECs, allowed to settle for 1–2 min, and immediately fixed with PFA. This was followed by immunolabeling of ICAM-1 and CX3CL1. Nuclei were labeled with Hoechst 33342 to identify CTLs.
(E) The synapses shown in (D) were analyzed using Fiji/ImageJ by determining intensity profiles along lines crossing the synapse from side to side. Yellow arrows highlight CTL nuclei. White circles highlight adherent CTLs. White rectangles indicate zoom-in area shown underneath. Dotted white line indicates an example line of measurement for plot profiles. Scale bar, 50μm.

From: Schoppmeyer R, van Steen ACI, Kempers L, Timmerman AL, Nolte MA, Hombrink P, van Buul JD.
The endothelial diapedesis synapse regulates transcellular migration of human T lymphocytes in a CX3CL1- and SNAP23-dependent manner.
Cell Rep. 2022 Jan 18;38(3):110243.
doi: 10.1016/j.celrep.2021.110243.
This image is from an open access article distributed under terms of a Creative Commons Attribution License.

Mouse anti Human CD54:Pacific Blue®

Product Type
Monoclonal Antibody
Clone
15.2
Isotype
IgG1
Specificity
CD54

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Product Code Applications Pack Size List Price Your Price Qty
MCA1615PB
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Mouse anti Human CD54 antibody, clone 15.2 recognizes the human CD54 cell surface antigen also known as intracellular Adhesion Molecule -1 (ICAM-1) or Major group rhinovirus receptor.

CD54 is expressed by many cells following activation by inflammatory mediators. It is a 505 amino acid with an additional 27 amino acid signal peptide ~90 kDa single pass type I transmembrane glycoprotein bearing 5 Ig-like C2-type domains.

Mouse anti Human CD54 antibody, clone 15.2 blocks CD54 function (Berendt et al. 1992). Mouse anti Human CD54 antibody, clone 15.2 binds to an epitope on the N-terminal Ig-like domain within a region designated the L43 loop (Chakravorty and Craig 2005).

Target Species
Human
Species Cross-Reactivity
Target SpeciesCross Reactivity
Pig
N.B. Antibody reactivity and working conditions may vary between species.
Product Form
Purified IgG conjugated to Pacific Blue® - liquid
Preparation
Purified IgG prepared by affinity chromatography on Protein A from tissue culture supernatant
Buffer Solution
Phosphate buffered saline
Preservative Stabilisers
0.09% sodium azide (NaN3)
1% bovine serum albumin
Immunogen
Human monocytes
Approx. Protein Concentrations
IgG concentration 0.05 mg/ml
Fusion Partners
Spleen cells from immunised BALB/c mice were fused with cells of the mouse Sp2/0-Ag14 myeloma cell line
Max Ex/Em
Fluorophore Excitation Max (nm) Emission Max (nm)
Pacific Blue® 410 455
Regulatory
For research purposes only
Guarantee
12 months from date of despatch
Acknowledgements
This product is provided under an intellectual property licence from Life Technologies Corporation. The transfer of this product is contingent on the buyer using the purchase product solely in research, excluding contract research or any fee for service research, and the buyer must not sell or otherwise transfer this product or its components for (a) diagnostic, therapeutic or prophylactic purposes; (b) testing, analysis or screening services, or information in return for compensation on a per-test basis; (c) manufacturing or quality assurance or quality control, or (d) resale, whether or not resold for use in research. For information on purchasing a license to this product for purposes other than as described above, contact Life Technologies Corporation, 5791 Van Allen Way, Carlsbad CA 92008 USA or outlicensing@thermofisher.com

This product is shipped at ambient temperature. It is recommended to aliquot and store at -20°C on receipt. When thawed, aliquot the sample as needed. Keep aliquots at 2-8°C for short term use (up to 4 weeks) and store the remaining aliquots at -20°C.

Avoid repeated freezing and thawing as this may denature the antibody. Storage in frost-free freezers is not recommended. This product is photosensitive and should be protected from light.

This product has been reported to work in the following applications. This information is derived from testing within our laboratories, peer-reviewed publications or personal communications from the originators. Please refer to references indicated for further information. For general protocol recommendations, please visit the antibody protocols page.
Application Name Verified Min Dilution Max Dilution
Flow Cytometry Neat
Where this product has not been tested for use in a particular technique this does not necessarily exclude its use in such procedures. Suggested working dilutions are given as a guide only. It is recommended that the user titrates the product for use in their own system using appropriate negative/positive controls.
Flow Cytometry
Use 10μl of the suggested working dilution to label 106 cells in 100μl

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Description Mouse IgG1 Negative Control:Pacific Blue®

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Description Human Seroblock

References for CD54 antibody

  1. Dransfield, I. et al. (1992) Interaction of leukocyte integrins with ligand is necessary but not sufficient for function.
    J Cell Biol. 116:1527-35.
  2. Berendt, A. et al. (1992) The binding site on ICAM-1 for plasmodium falciparum-infected erythrocytes overlaps, but is distinct from, the LFA-1- binding site.
    Cell. 68: 71-81.
  3. Urquhart, P. et al. (2007) Carbon monoxide-releasing molecules modulate leukocyte-endothelial interactions under flow.
    J Pharmacol Exp Ther. 321 (2): 656-62.
  4. Baratin, M. et al. (2007) Dissection of the role of PfEMP1 and ICAM-1 in the sensing of Plasmodium-falciparum-infected erythrocytes by natural killer cells.
    PLos One 2: e228.
  5. van Buul, J.D. et al. (2010) Inside-out regulation of ICAM-1 dynamics in TNF-alpha-activated endothelium.
    PLoS One 5: e11336.
  6. Diaz-Romero, J. et al. (2008) Immunophenotypic changes of human articular chondrocytes during monolayer culture reflect bona fide dedifferentiation rather than amplification of progenitor cells.
    J Cell Physiol. 214: 75-83.
  7. Di Lorenzo, A. et al. (2011) Endothelial reticulon-4B (Nogo-B) regulates ICAM-1-mediated leukocyte transmigration and acute inflammation.
    Blood. 117: 2284-95.
  8. Porter, J.C. and Hall, A. (2009) Epithelial ICAM-1 and ICAM-2 regulate the egression of human T cells across the bronchial epithelium.
    FASEB J. 23: 492-502.
    9. View The Latest Product References
  9. Corvaisier, M. et al. (2005) V gamma 9V delta 2 T cell response to colon carcinoma cells.
    J Immunol. 175: 5481-8.
  10. Horrocks, P. et al. (2005) PfEMP1 expression is reduced on the surface of knobless Plasmodium falciparum infected erythrocytes.
    J Cell Sci. 118: 2507-18.
  11. Lozanoska-Ochser, B. et al. (2008) Expression of CD86 on human islet endothelial cells facilitates T cell adhesion and migration.
    J Immunol. 181: 6109-16.
  12. Norling, L.V. et al. (2008) Inhibitory control of endothelial galectin-1 on in vitro and in vivo lymphocyte trafficking.
    FASEB J. 22 (3): 682-90.
  13. Baumer, Y. et al. (2011) Telomerase-based immortalization modifies the angiogenic/inflammatory responses of human coronary artery endothelial cells.
    Exp Biol Med (Maywood). 236: 692-700.
  14. Lask, A. et al. (2011) TCR-independent killing of B cell malignancies by anti-third-party CTLs: the critical role of MHC-CD8 engagement.
    J Immunol. 187 (4): 2006-14.
  15. Sommaggio, R. et al. (2012) Multiple Receptors Trigger Human NK Cell-Mediated Cytotoxicity against Porcine Chondrocytes.
    J Immunol. 188: 2075-83.
  16. Murphy, A.J. et al. (2013) Anti-inflammatory functions of apolipoprotein a-I and high-density lipoprotein are preserved in trimeric apolipoprotein a-I.
    J Pharmacol Exp Ther. 344: 41-9.
  17. Sumagin R et al. (2014) Transmigrated neutrophils in the intestinal lumen engage ICAM-1 to regulate the epithelial barrier and neutrophil recruitment.
    Mucosal Immunol. 7 (4): 905-15.
  18. Sugden SM et al. (2017) HIV-1 Vpu Downmodulates ICAM-1 Expression, Resulting in Decreased Killing of Infected CD4+ T Cells by NK Cells.
    J Virol. 91 (8): pii: e02442-16.
  19. Lennartz, F. et al. (2015) Mapping the Binding Site of a Cross-Reactive Plasmodium falciparum PfEMP1 Monoclonal Antibody Inhibitory of ICAM-1 Binding.
    J Immunol. 195 (7): 3273-83.
  20. Schoppmeyer, R. et al. (2022) The endothelial diapedesis synapse regulates transcellular migration of human T lymphocytes in a CX3CL1- and SNAP23-dependent manner.
    Cell Rep. 38 (3): 110243.
  21. Grönloh, M.L.B. et al. (2023) Endothelial transmigration hotspots limit vascular leakage through heterogeneous expression of ICAM-1.
    EMBO Rep. 24 (1): e55483.
  22. Grönloh, M.L.B. et al. (2023) Intercellular adhesion molecule 2 regulates diapedesis hotspots by allowing neutrophil crawling against the direction of flow.
    Vasc Biol. 5 (1): e230005.

Flow Cytometry

Immunofluorescence

Synonyms
ICAM-1
UniProt
P05362
Entrez Gene
ICAM1
GO Terms
GO:0007157 heterophilic cell-cell adhesion
GO:0001910 regulation of leukocyte mediated cytotoxicity
GO:0002291 T cell activation via T cell receptor contact with antigen bound to MHC molecule on antigen presenting cell
GO:0002693 positive regulation of cellular extravasation
GO:0004888 transmembrane receptor activity
GO:0005887 integral to plasma membrane
GO:0005178 integrin binding
GO:0005615 extracellular space
GO:0007159 leukocyte cell-cell adhesion
GO:0022614 membrane to membrane docking
GO:0046813 virion attachment, binding of host cell surface receptor
GO:0050776 regulation of immune response
GO:0050900 leukocyte migration
GO:0051856 adhesion to symbiont
GO:0060333 interferon-gamma-mediated signaling pathway

MCA1615PB

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