PE anti-human CD66b Antibody

Product Details

Verified Reactivity

Human

Reported Reactivity

Chimpanzee

Antibody Type

Monoclonal

Host Species

Mouse

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 PE under optimal conditions.

Concentration

Lot-specific (to obtain lot-specific concentration and expiration, please enter the lot number in our Certificate of Analysis online tool.)

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 - Community Verified

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.

Excitation Laser

Blue Laser (488 nm)
Green Laser (532 nm)/Yellow-Green Laser (561 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

For the use of this antibody in spatial biology (SB), we have partnered with Bruker Spatial Biology Biosciences for demonstration of this antibody on their next-generation ChipCytometry instrument called the CellScape™. The CellScape platform is an end-to-end solution for highly multiplexed spatial omics. Combining an advanced, purpose-built imaging system with easy-to-use fluidics for walk-away automation, the CellScape system will accelerate your exploration into the rapidly evolving field of spatial biology. More information on the the Bruker Spatial Biology CellScape and a complete list of our antibodies that have been demonstrated on the instrument can be found here.

Application References

(PubMed link indicates BioLegend citation)

1. Schlossman S, et al. Eds. 1995. Leucocyte Typing V. Oxford University Press. New York.
2. Kishimoto T, et al. Eds. 1997. Leucocyte Typing VI. Garland Publishing Inc. London.
3. Norling LV, et al. 2012. Arterioscler Thromb Vasc Biol. 32:1970. PubMed
4. Meinke P, et al. 2015. Neuroimmunol Discord. 25:127. PubMed
5. Radtke AJ, et al. 2020. Proc Natl Acad Sci USA. 117:33455-33465. (SB) PubMed
6. Radtke AJ, et al. 2022. Nat Protoc. 17:378-401. (SB) PubMed

Product Citations

1. Mirchandani AS, et al. 2022. Nat Immunol. 23:927. PubMed
2. Sehgal R, et al. 2022. Cells. 11: . PubMed
3. García-García A, et al. 2023. Bioact Mater. 24:174. PubMed
4. Zheng Y, et al. 2022. Proc Natl Acad Sci U S A. 119:e2121077119. PubMed
5. Lareau CA, et al. 2023. Nat Protoc. 18:1416. PubMed
6. Freund EC, et al. 2023. Methods Mol Biol. 2618:201. PubMed
7. Doloff JC, et al. 2023. Sci Adv. 9:eade9488. PubMed
8. O’Dea K, et al. 2016. PLoS One. 11:e0167801. PubMed
9. Klemm F, et al. 2020. Cell. 181(7):1643-1660.e17. PubMed
10. Barbu EA, et al. 2021. Bio Protoc. 11:e3927. PubMed
11. Butler AL, et al. 2019. Front Immunol. 10:1851. PubMed
12. Seman BG, et al. 2021. J Cell Sci. 134:. PubMed
13. Khan E, et al. 2016. Sci Rep. 6:38104. PubMed
14. Dong Y, et al. 2020. J Leukoc Biol. 108:1711. PubMed
15. Wu L, et al. 2018. Oncol Lett. 15:9507. PubMed
16. Mol S, et al. 2021. Int J Mol Sci. 22:. PubMed
17. Li X, et al. 2017. Stem Cell Res. 10.1016/j.scr.2017.03.014. PubMed
18. Crespo-Lessmann A, et al. 2017. J Asthma Allergy. 10:269. PubMed
19. Wang L, et al. 2022. Gut. Online ahead of print. PubMed
20. Canals Hernaez D, et al. 2022. Front Oncol. 12:856424. PubMed
21. Zhu YP et al. 2018. Cell reports. 24(9):2329-2341 . PubMed
22. Tähtinen S, et al. 2022. Nat Immunol. 23:532. PubMed
23. Schmidt T, et al. 2015. PLoS One. 10: 0132703. PubMed
24. Inglis H, et al. 2015. J Vis Exp. 97: 52484. PubMed
25. Dinh HQ, et al. 2020. Immunity. 53(2):319-334.e6. PubMed
26. Barbu EA, et al. 2020. Front Immunol. 1.385416667. PubMed
27. Miramón P, et al. 2012. PLoS One. 7:e52850. PubMed

RRID

AB_10550093 (BioLegend Cat. No. 305105)
AB_10550093 (BioLegend Cat. No. 305106)

Antigen Details

Structure

Ig superfamily, CEA antigen group, GPI-linked glycoprotein, 95-100 kD

Distribution

Granulocytes

Function

Cell adhesion, neutrophil activation

Ligand/Receptor

CD66c

Cell Type

Granulocytes, Neutrophils

Biology Area

Immunology

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

UniProt

View information about CD66b on UniProt.org

Documentation

• Technical Data Sheet (pdf)
• Certificate of Analysis
• Safety Data Sheet

Reviews

Related Protocols

• Cell Surface Flow Cytometry Staining Protocol

Related Pages & Pathways

Related FAQs

What type of PE do you use in your conjugates?

We use R-PE in our conjugates.

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.

Other Formats

Certificate of Analysis