- G10F5 (See other available formats)
- Regulatory Status
- VI MA81
- Other Names
- CD67, CGM6, NCA-95, CEACAM8
- Mouse IgM, κ
- Ave. Rating
- Submit a Review
- Product Citations
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CD66b is a 95-100 kD glycosylphosphatidylinositol (GPI)-linked protein also known as CD67, CGM6, and NCA-95. CD66b is a member of the immunoglobulin superfamily, carcinoembryonic antigen (CEA)-like subfamily. CD66b, expressed on granulocytes, has been reported to induce activation in neutrophils and to be involved in heterophilic adhesion with CD66c.Product Details
- Verified Reactivity
- Reported Reactivity
- Antibody Type
- Host Species
- Phosphate-buffered solution, pH 7.2, containing 0.09% sodium azide and BSA (origin USA)
- The antibody was purified by affinity chromatography and conjugated with Alexa Fluor® 647 under optimal conditions.
- 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.
FC - Quality tested
IHC-P - Verified
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. For immunohistochemical staining on formalin-fixed paraffin-embedded tissue sections, the suggested use of this reagent is 5.0 - 10 µg per ml. It is recommended that the reagent be titrated for optimal performance for each application.
* 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
Additional reported applications (for the relevant formats) include: immunohistochemical staining of acetone-fixed frozen, formalin-fixed paraffin-embedded tissue sections, and spatial biology (IBEX)5,6.
- 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).
(PubMed link indicates BioLegend citation)
- Schlossman S, et al. Eds. 1995. Leucocyte Typing V. Oxford University Press. New York.
- Kishimoto T, et al. Eds. 1997. Leucocyte Typing VI. Garland Publishing Inc. London.
- Norling LV, et al. 2012. Arterioscler Thromb Vasc Biol. 32:1970. PubMed
- Meinke P, et al. 2015. Neuroimmunol Discord. 25:127. PubMed
- Radtke AJ, et al. 2020. Proc Natl Acad Sci USA. 117:33455-33465. (SB) PubMed
- Radtke AJ, et al. 2022. Nat Protoc. 17:378-401. (SB) PubMed
AB_2563170 (BioLegend Cat. No. 305109)
AB_2563171 (BioLegend Cat. No. 305110)
- Ig superfamily, CEA antigen group, GPI-linked glycoprotein, 95-100 kD
- Cell adhesion, neutrophil activation
- Cell Type
- Granulocytes, Neutrophils
- Biology Area
- Molecular Family
- Adhesion Molecules, CD Molecules
- Antigen References
1. Kuijpers T, et al. 1993. J. Immunol. 151:4934.
2. Kuroki M, et al. 1992. J. Leuk. Biol. 52:551.
- Gene ID
- 1088 View all products for this Gene ID
- View information about CD66b on UniProt.org
- 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.
|FITC anti-human CD66b||G10F5||FC|
|Purified anti-human CD66b||G10F5||FC, IHC-P|
|Pacific Blue™ anti-human CD66b||G10F5||FC|
|PE anti-human CD66b||G10F5||FC|
|PerCP/Cyanine5.5 anti-human CD66b||G10F5||FC|
|Alexa Fluor® 647 anti-human CD66b||G10F5||FC, IHC-P, SB|
|Alexa Fluor® 700 anti-human CD66b||G10F5||FC|
|PE/Cyanine7 anti-human CD66b||G10F5||FC|
|APC anti-human CD66b||G10F5||FC|
|Biotin anti-human CD66b||G10F5||FC|
|PE/Dazzle™ 594 anti-human CD66b||G10F5||FC|
|Alexa Fluor® 594 anti-human CD66b||G10F5||IHC-P|
|APC/Cyanine7 anti-human CD66b||G10F5||FC|
|FITC anti-human CD66b||G10F5||FC|
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FITC anti-human CD66b
Purified anti-human CD66b
Pacific Blue™ anti-human CD66b
PE anti-human CD66b
PerCP/Cyanine5.5 anti-human CD66b
Alexa Fluor® 647 anti-human CD66b
Alexa Fluor® 700 anti-human CD66b
PE/Cyanine7 anti-human CD66b
APC anti-human CD66b
Biotin anti-human CD66b
PE/Dazzle™ 594 anti-human CD66b
Alexa Fluor® 594 anti-human CD66b
APC/Cyanine7 anti-human CD66b
FITC anti-human CD66b