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Spark Blue™ 574 anti-human CD16 (Flexi-Fluor™) Antibody

Pricing & Availability
Clone
3G8 (See other available formats)
Regulatory Status
RUO
Workshop
V NK80
Other Names
FcγRIII, Fc gamma receptor, Fc gamma receptor 3
Isotype
Mouse IgG1, κ
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Product Citations
publications
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Cat # Size Price Quantity Save
285021 50 µg £270
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Description

CD16 is known as low affinity IgG receptor III (FcγRIII). It is expressed as two distinct forms (CD16a and CD16b). CD16a (FcγRIIIA) is a 50-65 kD polypeptide-anchored transmembrane protein. It is expressed on the surface of NK cells, activated monocytes, macrophages, and placental trophoblasts in humans. CD16b (FcγRIIIB) is a 48 kD glycosylphosphatidylinositol (GPI)-anchored protein. Its extracellular domain is over 95% homologous to that of CD16a, and it is expressed specifically on neutrophils. CD16 binds aggregated IgG or IgG-antigen complex which functions in NK cell activation, phagocytosis, and antibody-dependent cell-mediated cytotoxicity (ADCC).

Product Details
Technical Data Sheet (pdf)

Product Details

Verified Reactivity
Human, Cynomolgus, Rhesus
Reported Reactivity
African Green, Baboon, Capuchin Monkey, Chimpanzee, Common Marmoset, Pigtailed Macaque, Sooty Mangabey, Squirrel Monkey
Antibody Type
Monoclonal
Host Species
Mouse
Immunogen
Human PMN cells
Formulation
Phosphate-buffered solution, pH 7.2, containing 0.09% sodium azide
Preparation
The antibody was purified by affinity chromatography and conjugated with Spark Blue™ 574 under optimal conditions.
Concentration
0.2 mg/mL
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
Recommended Usage
Flexi-Fluors™ are provided at a standard 0.2 mg/mL concentration. We recommend titrating this reagent to determine the optimal concentration for each application. For many flow cytometry applications, conjugated antibodies perform well at concentrations ranging from 0.03 to 1.0 µg per million cells in 100 µL. We recommend testing a range of concentrations starting from 10 µg/mL.

For example, make five 1:1 serial dilutions of the 0.2 mg/mL antibody. Add 5 µL of each dilution (including the undiluted antibody) to 100 µL of cells (at 107 cells/mL) to test six concentrations -- 1.0, 0.5, 0.25, 0.125, 0.06, and 0.03 µg per million cells in 100 µL volume. Compare staining patterns or create a titration curve using the MFI or staining index to determine the optimal concentration.
Excitation Laser
Blue Laser (488 nm)
Application Notes

The 3G8 antibody clone blocks neutrophil phagocytosis and stimulates NK cell proliferation. It has been reported that this clone interacts with the FcγRIIa and FcγRIIIb receptors causing neutrophil activation and aggregation18. Due to this phenomenon staining in whole blood may cause a reduction in the number of granulocytes or alter their scatter profile.

Additional reported applications (for the relevant formats) include: immunohistochemical staining of acetone-fixed frozen tissue sections6, immunoprecipitation3, stimulation of NK cell proliferation4, blocking of phagocytosis5, and blocking of immunoglobulin binding to FcγRIII7,8. The Ultra-LEAF™ purified antibody (Endotoxin < 0.01 EU/µg, Azide-Free, 0.2 µm filtered) is recommended for functional assays (Cat. No. 302049, 302050, 302057, 302058).

Additional Product Notes

For more information about Flexi-Fluors™, visit our Flexi-Fluor™ page and review FAQs associated with this product line.

Application References

(PubMed link indicates BioLegend citation)
  1. Knapp W, et al. Eds. 1989. Leucocyte Typing IV. Oxford University Press. New York.
  2. Schlossman S, et al. Eds. 1995. Leucocyte Typing V. Oxford University Press. New York.
  3. Edberg J, et al. 1997. J. Immunol. 159:3849. (IP)
  4. Hoshino S, et al. 1991. Blood 78:3232. (Stim)
  5. Tamm A, et al. 1996. Immunol. 157:1576. (Block)
  6. Da Silva DM, et al. 2001. Int. Immunol. 13:633. (IHC)
  7. Holl V, et al. 2004. J. Immunol. 173:6274. (Block)
  8. Hober D, et al. 2002. J. Gen. Virol. 83:2169. (Block)
  9. Brainard DM, et al. 2009. J. Virol. 83:7305. PubMed
  10. Smed-Sörensen A, et al. 2008. Blood 111:5037. (Block) PubMed
  11. Timmerman KL, et al. 2008. J. Leukoc. Biol. 84:1271. (FC) PubMed
  12. Yoshino N, et al. 2000. Exp. Anim. (Tokyo) 49:97. (FC)
  13. Rout N, et al. 2010. PLoS One 5:e9787. (FC)
  14. Kim WK, et al. 2006. Am. J. Pathol. 168:822. (FC)
  15. Boltz A, et al. 2011. J. Biol Chem. 286:21896. PubMed
  16. Wu Z, et al. 2013. J. Virol. 87:7717. PubMed
  17. Peterson VM, et al. 2017. Nat. Biotechnol. 35:936. (PG)
  18. Vossebeld PJ, et al. 1997. Biochem J. 323:87-94 (Stim)
RRID
AB_3106286 (BioLegend Cat. No. 285021)

Antigen Details

Structure
Ig superfamily, transmembrane form (50-65 kD) or GPI-linked form (48 kD)
Distribution

NK cells, activated monocytes, macrophages, neutrophils

Function
Low affinity IgG Fc receptor, phagocytosis, ADCC
Ligand/Receptor
Aggregated IgG, IgG-antigen complex
Cell Type
Dendritic cells, Macrophages, Monocytes, Neutrophils, NK cells
Biology Area
Immunology, Innate Immunity
Molecular Family
CD Molecules, Fc Receptors
Antigen References

1. Fleit H, et al. 1982. P. Natl. Acad. Sci. USA 79:3275.
2. Stroncek D, et al. 1991. Blood 77:1572.
3. Wirthmueller U, et al. 1992. J. Exp. Med. 175:1381.

Gene ID
2214 View all products for this Gene ID
UniProt
View information about CD16 on UniProt.org

Related FAQs

Is our human Trustain FcX™ (cat# 422302) compatible with anti human CD16, CD32 and CD64 clones 3G8, FUN-2 and 10.1 respectively?

Yes

What are Flexi-Fluors?

Flexi-Fluors are rapidly made-to-order conjugated antibodies. The technology, manufacturing processes, and specifications used to create Flexi-Fluors are the same as our regular catalog products. However, the optimal concentration and performance of each Flexi-Fluor must be determined by the customer. 

How quickly will I receive my order?

We aim to ship Flexi-Fluors within 2-3 weeks of receipt of your order. However, depending on your location, shipping times may vary. 

How are Flexi-Fluors different from regular catalog products?

Flexi-Fluors are made on demand, specifically for you. Flexi-Fluors are manufactured using the same high-quality standards, and specifications as other catalog products. For faster delivery, Flexi-Fluors are not tested by flow cytometry to determine optimal concentrations or evaluate performance. This testing needs to be performed by the customer.

How do I determine the optimal concentration for using my Flexi-Fluor? How should I titrate my antibody?

Flexi-Fluors are provided at a standard 0.2 mg/mL concentration. We recommend that you titrate your antibody to determine the optimal concentration to use for your application. For many flow cytometry applications, conjugated antibodies perform well at concentrations ranging from 0.03 to 1.0 µg per million cells in 100 µL volume. We recommend that you test a range of concentrations starting from 10 µg/mL. 

 

For example, make five 1:1 serial dilutions of your 0.2 mg/mL antibody.  Add 5 µL of each dilution (including the undiluted antibody) to 100 µL of cells (at 107 cells/ml) to test six concentrations - 1.0, 0.5, 0.25, 0.125, 0.06, and 0.03 µg per million cells in 100 µL volume. Compare staining patterns or create a titration curve using the MFI or staining index to determine the optimal concentration. 

I can’t find the antibody-dye combination that I need. When will it be available?

We continuously update our catalog, introducing scores of new products every month. Please get in touch with our Technical Service team for an update on new products or recommendations for suitable alternatives to complete your panel. Or contact Custom Solutions to inquire about our affordable custom conjugation services. 

I need help to validate the performance of my Flexi-Fluor. Who should I contact?

Please get in touch with Technical Service for assistance. 

Can I order more than 50 μg of a Flexi-Fluor?

Yes, you can order multiple vials of the same Flexi-Fluor products.  We cannot guarantee, however, that these vials will be bottled from the same lot. For bulk single-lot orders, contact our Custom Solutions team.

What is the expiration date of my Flexi-Fluor?

Expiration dates can be found on the vial label or by using our CoA lookup tool. 

Other Formats

View All CD16 Reagents Request Custom Conjugation
Description Clone Applications
APC anti-human CD16 3G8 FC
Biotin anti-human CD16 3G8 FC
FITC anti-human CD16 3G8 FC
Brilliant Violet 711™ anti-human CD16 3G8 FC
PE anti-human CD16 3G8 FC
PE/Cyanine5 anti-human CD16 3G8 FC
Purified anti-human CD16 3G8 FC,CyTOF®,Block,IHC-F,IP,Stim
APC/Cyanine7 anti-human CD16 3G8 FC
PE/Cyanine7 anti-human CD16 3G8 FC
Alexa Fluor® 488 anti-human CD16 3G8 FC
Alexa Fluor® 647 anti-human CD16 3G8 FC
Pacific Blue™ anti-human CD16 3G8 FC
Alexa Fluor® 700 anti-human CD16 3G8 FC
PerCP/Cyanine5.5 anti-human CD16 3G8 FC
PerCP anti-human CD16 3G8 FC
Brilliant Violet 421™ anti-human CD16 3G8 FC
Brilliant Violet 570™ anti-human CD16 3G8 FC
Brilliant Violet 605™ anti-human CD16 3G8 FC
Brilliant Violet 650™ anti-human CD16 3G8 FC
Brilliant Violet 785™ anti-human CD16 3G8 FC
Brilliant Violet 510™ anti-human CD16 3G8 FC
Ultra-LEAF™ Purified anti-human CD16 3G8 FC,CyTOF®,Block,IHC-F,IP,Stim
Purified anti-human CD16 (Maxpar® Ready) 3G8 FC,CyTOF®
PE/Dazzle™ 594 anti-human CD16 3G8 FC
APC/Fire™ 750 anti-human CD16 3G8 FC
PE anti-human CD16 3G8 FC
APC anti-human CD16 3G8 FC
Pacific Blue™ anti-human CD16 3G8 FC
PE/Dazzle™ 594 anti-human CD16 3G8 FC
TotalSeq™-A0083 anti-human CD16 3G8 PG
TotalSeq™-B0083 anti-human CD16 3G8 PG
TotalSeq™-C0083 anti-human CD16 3G8 PG
PE/Cyanine7 anti-human CD16 3G8 FC
PE/Fire™ 640 anti-human CD16 3G8 FC
FITC anti-human CD16 3G8 FC
Spark YG™ 581 anti-human CD16 3G8 FC
APC/Fire™ 750 anti-human CD16 3G8 FC
TotalSeq™-D0083 anti-human CD16 3G8 PG
APC/Fire™ 810 anti-human CD16 3G8 FC
GMP APC anti-human CD16 3G8 FC
GMP PE/Dazzle™ 594 anti-human CD16 3G8 FC
GMP PE anti-human CD16 3G8 FC
Spark Red™ 718 anti-human CD16 3G8 FC
GMP Pacific Blue™ anti-human CD16 3G8 FC
GMP FITC anti-human CD16 3G8 FC
Spark Blue™ 515 anti-human CD16 3G8 FC
Spark UV™ 387 anti-human CD16 3G8 FC
PerCP/Cyanine5.5 anti-human CD16 3G8 FC
GMP PE/Cyanine7 anti-human CD16 3G8 FC
GMP APC/Fire™ 750 anti-human CD16 3G8 FC
Brilliant Violet 750™ anti-human CD16 3G8 FC
Spark Blue™ 550 anti-human CD16 3G8 FC
GMP PerCP/Cyanine5.5 anti-human CD16 3G8 FC
Spark YG™ 593 anti-human CD16 3G8 FC
Spark NIR™ 685 anti-human CD16 3G8 FC
Spark Violet™ 500 anti-human CD16 3G8 FC
Spark Blue™ 574 anti-human CD16 (Flexi-Fluor™) 3G8 FC
Spark PLUS UV395™ anti-human CD16 3G8 FC
Go To Top Version: 1    Revision Date: 02/06/2024

For Research Use Only. Not for diagnostic or therapeutic use.

 

This product is supplied subject to the terms and conditions, including the limited license, located at www.biolegend.com/terms) ("Terms") and may be used only as provided in the Terms. Without limiting the foregoing, BioLegend products may not be used for any Commercial Purpose as defined in the Terms, resold in any form, used in manufacturing, or reverse engineered, sequenced, or otherwise studied or used to learn its design or composition without express written approval of BioLegend. Regardless of the information given in this document, user is solely responsible for determining any license requirements necessary for user’s intended use and assumes all risk and liability arising from use of the product. BioLegend is not responsible for patent infringement or any other risks or liabilities whatsoever resulting from the use of its products.

 

BioLegend, the BioLegend logo, and all other trademarks are property of BioLegend, Inc. or their respective owners, and all rights are reserved.

 

8999 BioLegend Way, San Diego, CA 92121 www.biolegend.com
Toll-Free Phone: 1-877-Bio-Legend (246-5343) Phone: (858) 768-5800 Fax: (877) 455-9587

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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