Ultra-LEAF™ Purified anti-mouse CD16/32 Antibody

Product Details

Verified Reactivity

Mouse

Antibody Type

Monoclonal

Host Species

Rat

Immunogen

Sorted pre-B cells

Formulation

0.2 µm filtered in phosphate-buffered solution, pH 7.2, containing no preservative.

Endotoxin Level

Less than 0.01 EU/µg of the protein (< 0.001 ng/µg of the protein) as determined by the LAL test.

Preparation

The Ultra-LEAF™ (Low Endotoxin, Azide-Free) antibody was purified by affinity chromatography.

Concentration

The antibody is bottled at the concentration indicated on the vial, typically between 2 mg/mL and 3 mg/mL. Older lots may have also been bottled at 1 mg/mL. 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. This Ultra-LEAF™ solution contains no preservative; handle under aseptic conditions.

Application

FC - Quality tested
IP, Block - 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 ≤ 1.0 µg per million cells in 100 µl volume or 100 µl of whole blood.  It is recommended that the reagent be titrated for optimal performance for each application.

Application Notes

Clone 93 can be used for blocking of CD16/CD32 interactions with the Fc domain of immunoglobulins, but is not the same clone as 2.4G2.

The 93 mAb is specific to the common epitope of CD16/CD32. Additional reported applications (for the relevant formats) include: immunoprecipitation¹ and blocking of Fc-mediated reactions in functional studies^2,4,23. It is useful for blocking non-specific binding of immunoglobulin to Fc receptors. For blocking of Fc receptors in flow cytometric analysis, pre-incubate the cells with purified anti-CD16/CD32 antibody (=1.0 µg per 10⁶ cells in 100 µL volume) for 5-10 minutes on ice prior to immunostaining. For highly sensitive assays, we recommend Ultra-LEAF™ purified antibody (Cat. No. 101330) (Endotoxin <0.01 EU/µg, Azide-Free, 0.2 µm filtered).

Application References

(PubMed link indicates BioLegend citation)

1. Personal communication (IP)
2. Oliver AM, et al. 1999. Hybridoma 18:113. (Block)
3. Brummel R and Lenert P. 2005. J. Immunol. 174:2429.
4. Terrazas LI, et al. 2005. Int. J. Parasitol. 35:1349. (Block)
5. Clements JL, et al. 2006. J. Immunol. 177:905.
6. Mohamed W, et al. 2010. Infect Immun. 78:3306. PubMed
7. Ouchi T, et al. 2011. J. Exp Med. 208:2607. PubMed
8. Kmieciak M, et al. 2011. J. Vis. Exp. 47:2381. PubMed
9. Yamazaki S, et al. 2012. PLoS One. 7:e51665. PubMed
10. Li J, et al. 2012. Arthritis Rheum. 64:1098. PubMed
11. Azuma M, et al. 2012. Oncoimmunology. 1:581. PubMed
12. Koon HW, et al. 2013. J. Vis. Exp. 68:4208. PubMed
13. Hegde VL, et al. 2013. J Biol Chem. 288:36810. PubMed
14. Huang J, et al. 2013. J. Immunol Methods. 387:254. PubMed
15. Dutow P, et al. 2014. J Infect Dis. PubMed
16. Fan Y, et al. 2014. J Exp Med. 211:313. PubMed
17. Huang HN, et al. 2014. Antimicrob Agents Chemother. 58:1538. PubMed
18. Takei S, et al. 2014. Vaccine. 32:3066. PubMed
19. Richardson ML, et al. 2014. PLoS Negl Trop Dis. 8:2825. PubMed
20. Cekanaviciute E, et al. 2014. J Immunol. 193:139. PubMed
21. Kimura T, et al. 2014. Int Immunol. 26:697. PubMed
22. Everad A, et al. 2014. Nat Commun. 5:5648. PubMed
23. Cenci E, et al. 2006. J. Leuko. Biol. 79(1):40-5. (Block)

Product Citations

1. Dong MB, et al. 2020. Cell. 178(5):1189-1204.e23.. PubMed
2. Liang Y, et al. 2021. Cell Death Differ. 28:2728. PubMed
3. Miranda K, et al. 2022. iScience. 25:104994. PubMed
4. Kraemer AN, et al. 2022. Front Immunol. 13:933191. PubMed
5. Yang L, et al. 2023. Sci Rep. 13:1827. PubMed
6. van Os BW, et al. 2023. Eur Heart J Open. 3:oead013. PubMed
7. van Os BW, et al. 2023. Front Cardiovasc Med. 10:1171764. PubMed
8. Matsudaira T, et al. 2023. Commun Biol. 6:665. PubMed
9. Nair N, et al. 2020. Curr Protoc Microbiol. 59:e127. PubMed
10. Schmidleithner L et al. 2019. Immunity. 50(5):1232-1248 . PubMed
11. Petursdottir D, et al. 2017. Front Immunol. . 10.3389/fimmu.2017.01699. PubMed
12. Xie Y, et al. 2020. Journal of Neurochemistry. 152(3):368-380.. PubMed
13. Bartleson JM, et al. 2020. Nat Immunol. 1384:21. PubMed
14. Lercher A, et al. 2019. Immunity. 51:1074. PubMed
15. Pfenninger P, et al. 2022. Front Immunol. 13:777113. PubMed
16. Guérin MV, et al. 2019. Nat Commun. 10:4131. PubMed
17. Guo P, et al. 2021. J Immunol. 207:408. PubMed
18. Li D, et al. 2022. Emerg Microbes Infect. 11:2248. PubMed
19. Arensman MD, et al. 2020. Oncoimmunology. 9:1800162. PubMed
20. Le TM et al. 2018. Journal of neurochemistry. 145(2):139-153 . PubMed
21. Fox A, et al. 2020. Current Protocols in Cytometry. 93(1):e74. PubMed
22. Schäfer AL, et al. 2021. Front Immunol. 12:696810. PubMed
23. Trindade BC, et al. 2021. Immunity. 54:2273. PubMed
24. Amend A, et al. 2021. Int J Mol Sci. 22:. PubMed
25. Tong Y, et al. 2018. EBioMedicine. 39:132. PubMed

RRID

AB_2561482 (BioLegend Cat. No. 101329)
AB_2561482 (BioLegend Cat. No. 101330)
AB_2561482 (BioLegend Cat. No. 101339)
AB_2561482 (BioLegend Cat. No. 101340)
AB_2561482 (BioLegend Cat. No. 101341)
AB_2561482 (BioLegend Cat. No. 101342)

Antigen Details

Structure

Ig superfamily, 40-60 kD

Distribution

B cells, monocyte/macrophages, NK cells, neutrophils, mast cells, dendritic cells

Function

Low affinity receptors for IgG

Ligand/Receptor

IgG

Cell Type

B cells, Dendritic cells, Macrophages, Mast cells, Monocytes, Neutrophils, NK cells

Biology Area

Immunology, Innate Immunity

Molecular Family

CD Molecules, Fc Receptors

Antigen References

1. Barclay AN, et al. 1997. The Leukocyte Antigen FactsBook Academic Press.
2. Unkeless JC. 1989. J. Clin. Invest. 83:355.
3. Qiu WQ, et al. 1990. Science 248:732.

Gene ID

14130 View all products for this Gene ID 14131 View all products for this Gene ID

UniProt

View information about CD16/32 on UniProt.org

Documentation

• Technical data sheet
• Certificate of Analysis
• Safety Data Sheet

Reviews

Related Protocols

• Cell Surface Flow Cytometry Staining Protocol

Related Pages & Pathways

Related FAQs

Do you guarantee that your antibodies are totally pathogen free?

BioLegend does not test for pathogens in-house aside from the GoInVivo™ product line. However, upon request, this can be tested on a custom basis with an outside, independent laboratory.

Does BioLegend test each Ultra-LEAF™ antibody by functional assay?

No, BioLegend does not test Ultra-LEAF™ antibodies by functional assays unless otherwise indicated. Due to the possible complexities and variations of uses of biofunctional antibodies in different assays and because of the large product portfolio, BioLegend does not currently perform functional assays as a routine QC for the antibodies. However, we do provide references in which the antibodies were used for functional assays and we do perform QC to verify the specificity and quality of the antibody based on our strict specification criteria.

Does BioLegend test each Ultra-LEAF™ antibody for potential pathogens?

No, BioLegend does not test for pathogens in-house unless otherwise indicated.  However, we can recommend an outside vendor to perform this testing as needed.

Have you tested this Ultra-LEAF™ antibody for in vivo or in vitro applications?

We don't test our antibodies for in vivo or in vitro applications unless otherwise indicated. Depending on the product, the TDS may describe literature supporting usage of a particular product for bioassay. It may be best to further consult the literature to find clone specific information.

Other Formats

Certificate of Analysis