Alexa Fluor® 647 anti-human CD54 Antibody

Pricing & Availability
Clone
HA58 (See other available formats)
Regulatory Status
RUO
Workshop
HCDM listed
Other Names
ICAM-1, Ly-47
Isotype
Mouse IgG1, κ
Ave. Rating
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Product Citations
publications
1_HA58_A647_CD54_Antibody_072817
Human peripheral blood lymphocytes were stained with CD54 (clone HA58) Alexa Fluor® 647 (filled histogram) or mouse IgG1, κ Alexa Fluor® 647 isotype control (open histogram).
  • 1_HA58_A647_CD54_Antibody_072817
    Human peripheral blood lymphocytes were stained with CD54 (clone HA58) Alexa Fluor® 647 (filled histogram) or mouse IgG1, κ Alexa Fluor® 647 isotype control (open histogram).
  • 2_13_Human_Spleen_CD21_CD54
    Confocal image of human spleen sample acquired using the IBEX method of highly multiplexed antibody-based imaging: CD21 (red) in Cycle 1 and CD54 (blue) in Cycle 1. Tissues were prepared using ~1% (vol/vol) formaldehyde and a detergent. Following fixation, samples are immersed in 30% (wt/vol) sucrose for cryoprotection. Images are courtesy of Drs. Andrea J. Radtke and Ronald N. Germain of the Center for Advanced Tissue Imaging (CAT-I) in the National Institute of Allergy and Infectious Diseases (NIAID, NIH).
  • 3_17_Human_Liver_CD54_CD138
    Confocal image of human liver sample acquired using the IBEX method of highly multiplexed antibody-based imaging: CD54 (cyan) in Cycle 1 and CD138 (purple) in Cycle 2. Tissues were prepared using ~1% (vol/vol) formaldehyde and a detergent. Following fixation, samples are immersed in 30% (wt/vol) sucrose for cryoprotection. Images are courtesy of Drs. Andrea J. Radtke and Ronald N. Germain of the Center for Advanced Tissue Imaging (CAT-I) in the National Institute of Allergy and Infectious Diseases (NIAID, NIH).
  • 4_27_Human_Jejunum_Hoechst_CD54_Vimentin
    Confocal image of human jejunum sample acquired using the IBEX method of highly multiplexed antibody-based imaging: Hoechst (blue) in Cycle 1, CD54 (green) in Cycle 1, and Vimentin (magenta) in Cycle 6. Tissues were prepared using ~1% (vol/vol) formaldehyde and a detergent. Following fixation, samples are immersed in 30% (wt/vol) sucrose for cryoprotection. Images are courtesy of Drs. Andrea J. Radtke and Ronald N. Germain of the Center for Advanced Tissue Imaging (CAT-I) in the National Institute of Allergy and Infectious Diseases (NIAID, NIH).
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353113 25 tests $110
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353114 100 tests $248
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Description

CD54 is a 85-110 kD type I transmembrane protein also known as ICAM-1. It is expressed on activated endothelial cells, high endothelial venules, T and B cells, monocytes/macrophages, granulocytes, and dendritic cells. The expression of ICAM-1 can be released from the cell surface. CD54 plays a role in cellular adhesion and is involved in inflammation and leukocyte extravasation. CD54 has also been shown to be the major cellular receptor for rhinovirus. ICAM-1 binds to CD11a/CD18 (LFA-1), CD11b/CD18 (Mac-1), CD11c/CD18 (p150, 95) as well as hyaluronan and fibrinogen.

Product Details
Technical data sheet

Product Details

Verified Reactivity
Human
Antibody Type
Monoclonal
Host Species
Mouse
Immunogen
Colonic cancer BM314 cells
Formulation
Phosphate-buffered solution, pH 7.2, containing 0.09% sodium azide and BSA (origin USA)
Preparation
The antibody was purified by affinity chromatography and conjugated with Alexa Fluor® 647 under optimal conditions.
Concentration
Lot-specific (to obtain lot-specific concentration, please enter the lot number in our Concentration and Expiration Lookup or Certificate of Analysis online tools.)
Storage & Handling
The antibody solution should be stored undiluted between 2°C and 8°C, and protected from prolonged exposure to light. Do not freeze.
Application

FC - Quality tested
SB - Reported in the literature, not verified in house

Recommended Usage

Each lot of this antibody is quality control tested by immunofluorescent staining with flow cytometric analysis. For flow cytometric staining, the suggested use of this reagent is 5 µl per million cells in 100 µl staining volume or 5 µl per 100 µl of whole blood.

* Alexa Fluor® 647 has a maximum emission of 668 nm when it is excited at 633 nm / 635 nm.


Alexa Fluor® and Pacific Blue™ are trademarks of Life Technologies Corporation.

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Excitation Laser
Red Laser (633 nm)
Application Notes

Clone HA58 recognizes an epitope located in the extracellular D1 domain of CD543. Additional reported applications (for the relevant formats) include: spatial biology (IBEX)4,5.

Additional Product Notes

Iterative Bleaching Extended multi-pleXity (IBEX) is a fluorescent imaging technique capable of highly-multiplexed spatial analysis. The method relies on cyclical bleaching of panels of fluorescent antibodies in order to image and analyze many markers over multiple cycles of staining, imaging, and, bleaching. It is a community-developed open-access method developed by the Center for Advanced Tissue Imaging (CAT-I) in the National Institute of Allergy and Infectious Diseases (NIAID, NIH).

Application References

(PubMed link indicates BioLegend citation)
  1. Tsujisaki M, et al. 1991. Clin. Exp. Immunol. 85:3.
  2. Kanwar JR, et al. 2003. Cancer Gene Ther. 10:468.
  3. Kohka H, et al. 1998. J. Leukoc. Biol. 64:519.
  4. Radtke AJ, et al. 2020. Proc Natl Acad Sci USA. 117:33455-33465. (SB) PubMed
  5. Radtke AJ, et al. 2022. Nat Protoc. 17:378-401. (SB) PubMed
RRID
AB_2715941 (BioLegend Cat. No. 353113)
AB_2715942 (BioLegend Cat. No. 353114)

Antigen Details

Structure
Type I membrane protein, Ig superfamily, 85-110 kD
Distribution

Endothelial cells, T cells and B cells, monocytes/macrophages, granulocytes, and dendritic cells

Cell Type
B cells, Dendritic cells, Endothelial cells, Granulocytes, Macrophages, Mesenchymal Stem Cells, Monocytes, T cells
Biology Area
Cell Adhesion, Cell Biology, Costimulatory Molecules, Immunology, Neuroscience, Neuroscience Cell Markers, Stem Cells
Molecular Family
Adhesion Molecules, CD Molecules
Antigen References

1. Voraberger G, et al. 1991. J. Immunol. 147:2777.
2. Staunton DE, et al. 1988. Cell 52:925.
3. Greve JM, et al. 1989. Cell 56:839.

Gene ID
3383 View all products for this Gene ID
UniProt
View information about CD54 on UniProt.org

Related FAQs

If an antibody clone has been previously successfully used in IBEX in one fluorescent format, will other antibody formats work as well?

It’s likely that other fluorophore conjugates to the same antibody clone will also be compatible with IBEX using the same sample fixation procedure. Ultimately a directly conjugated antibody’s utility in fluorescent imaging and IBEX may be specific to the sample and microscope being used in the experiment. Some antibody clone conjugates may perform better than others due to performance differences in non-specific binding, fluorophore brightness, and other biochemical properties unique to that conjugate.

Will antibodies my lab is already using for fluorescent or chromogenic IHC work in IBEX?

Fundamentally, IBEX as a technique that works much in the same way as single antibody panels or single marker IF/IHC. If you’re already successfully using an antibody clone on a sample of interest, it is likely that clone will have utility in IBEX. It is expected some optimization and testing of different antibody fluorophore conjugates will be required to find a suitable format; however, legacy microscopy techniques like chromogenic IHC on fixed or frozen tissue is an excellent place to start looking for useful antibodies.

Are other fluorophores compatible with IBEX?

Over 18 fluorescent formats have been screened for use in IBEX, however, it is likely that other fluorophores are able to be rapidly bleached in IBEX. If a fluorophore format is already suitable for your imaging platform it can be tested for compatibility in IBEX.

The same antibody works in one tissue type but not another. What is happening?

Differences in tissue properties may impact both the ability of an antibody to bind its target specifically and impact the ability of a specific fluorophore conjugate to overcome the background fluorescent signal in a given tissue. Secondary stains, as well as testing multiple fluorescent conjugates of the same clone, may help to troubleshoot challenging targets or tissues. Using a reference control tissue may also give confidence in the specificity of your staining.

How can I be sure the staining I’m seeing in my tissue is real?

In general, best practices for validating an antibody in traditional chromogenic or fluorescent IHC are applicable to IBEX. Please reference the Nature Methods review on antibody based multiplexed imaging for resources on validating antibodies for IBEX.

Go To Top Version: 2    Revision Date: 04/19/2022

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*These products may be covered by one or more Limited Use Label Licenses (see the BioLegend Catalog or our website, www.biolegend.com/ordering#license). BioLegend products may not be transferred to third parties, resold, modified for resale, or used to manufacture commercial products, reverse engineer functionally similar materials, or to provide a service to third parties without written approval of BioLegend. By use of these products you accept the terms and conditions of all applicable Limited Use Label Licenses. Unless otherwise indicated, these products are for research use only and are not intended for human or animal diagnostic, therapeutic or commercial use.

 

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This data display is provided for general comparisons between formats.
Your actual data may vary due to variations in samples, target cells, instruments and their settings, staining conditions, and other factors.
If you need assistance with selecting the best format contact our expert technical support team.

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