Alexa Fluor® 488 anti-mouse/human CD11b Antibody

Product Details

Verified Reactivity

Mouse, Human, Cross-Reactivity: Chimpanzee, Baboon, Cynomolgus, Rhesus, Rabbit (Lapine)

Antibody Type

Monoclonal

Host Species

Rat

Immunogen

C57BL/10 splenocytes

Formulation

Phosphate-buffered solution, pH 7.2, containing 0.09% sodium azide.

Preparation

The antibody was purified by affinity chromatography and conjugated with Alexa Fluor® 488 under optimal conditions.

Concentration

0.5 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 - Quality tested
IHC-F, 3D IHC - 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 ≤ 0.25 µg per 10⁶ cells in 100 µl volume. For immunohistochemical staining on frozen tissue sections, the suggested use of this reagent is 2.5 - 10 µg per ml. For 3D immunohistochemistry on formalin-fixed tissues, a concentration of 5.0 µg/mL is suggested. It is recommended that the reagent be titrated for optimal performance for each application.

* Alexa Fluor® 488 has a maximum emission of 519 nm when it is excited at 488 nm.

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

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Excitation Laser

Blue Laser (488 nm)

Application Notes

Clone M1/70 has been verified for immunocytochemistry (ICC) and frozen immunohistochemistry (IHC-F).

Additional reported applications (for relevant formats of this clone) include: immunoprecipitation^1,4, in vitro blocking^3,9,12, depletion^2,8, immunofluorescence microscopy^6,7,10, immunohistochemistry of acetone-fixed frozen sections^5,11-13, and spatial biology (IBEX)^35,36. For in vivo studies or highly sensitive assays, we recommend Ultra-LEAF™ purified antibody (Endotoxin < 0.01 EU/µg, Azide-Free, 0.2 µm filtered) (Cat. No. 101248).

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. Springer T, et al. 1978. Eur. J. Immunol. 8:539. (IP)
2. Ault K and Springer T. 1981. J. Immunol. 126:359. (Deplete)
3. Springer TA, et al. 1982. Immunol. Rev. 68:171. (Block)
4. Ho MK and Springer TA. 1983. J. Biol. Chem. 258:2766. (IP)
5. Flotte TJ, et al. 1983. Am. J. Pathol. 111:112. (IHC)
6. Noel GJ, et al. 1990. J. Clin. Invest. 85:208. (IF)
7. Allen LA and Aderem A. 1996. J. Exp. Med. 184:627 (IF)
8. D'Amico A and Wu L. 2003. J. Exp. Med. 198:293. (Deplete)
9. Brickson SJ, et al. 2003. Appl Physiol. 95:969. (Block)
10. Clatworthy MR and Smith KG. 2004. J. Exp. Med. 199:717. (IF)
11. Hata H, et al. 2004. J. Clin. Invest. 114:582. (IHC)
12. Zhang Y, et al. 2002. J. Immunol. 168:3088. (IHC)
13. Iwasaki A and Kelsall BL. 2001. J. Immunol. 166:4884 (IHC, FC)
14. Tailleux L. 2003. J. Exp. Med. 197:121. (Block, FC)
15. Olver S, et al. 2006. Cancer Research 66:571. (FC)
16. Tan SL, et al. 2006. J. Immunol. 176:2872. (FC) PubMed
17. Ponomarev ED, et al. 2006. J. Immunol. 176:1402. (FC)
18. Dzhagalov I, et al. 2007. Blood 109:1620. (FC)
19. Fazilleau N, et al. 2007. Nature Immunol. 8:753.
20. Rasmussen JW, et al. 2006. Infect. Immun.74:6590. PubMed
21. Napimoga MH, et al. 2008. J. Immunol. 180:609. PubMed
22. Elqaraz-Carmon V, et al. 2008. J. Lipid. Res. 49:1894. PubMed
23. Kim DD, et al. 2008. Blood 112:1109. PubMed
24. Guo Y, et al. 2008. Blood 112:480. PubMed
25. Norian LA, et al. 2009. Cancer Res. 69:3086. (FC) PubMed
26. Baumgartner CK, et al. 2010. J. Immunol. 184:573. PubMed
27. Charles N, et al. 2010. Nat. Med. 16:701. (FC) PubMed
28. Whiteland J, et al. 1995. J. Histochem. Cytochem. 43:313. (IHC)
29. Weber GF, et al. 2014. J Exp Med. 211:1243. PubMed
30. Ashok A, et al. 2015. Toxicol Sci. 143:64. PubMed
31. Price PJ, et al. 2015. J Immunol. 194:1164. PubMed
32. Doni A, et al. 2015. J Exp Med. 212:905. PubMed
33. Ferreira R, et al. 2016. J Infect Dis. 213: 669 - 673. PubMed
34. Peterson VM, et al. 2017. Nat. Biotechnol. 35:936. (PG)
35. Radtke AJ, et al. 2020. Proc Natl Acad Sci U S A. 117:33455-65. (SB) PubMed
36. Radtke AJ, et al. 2022. Nat Protoc. 17:378-401. (SB) PubMed

Product Citations

1. Zhang H, et al. 2017. Leukemia. 10.1128/mBio.00226-17. PubMed
2. Mooney C, et al. 2017. International Journal of Molecular Sciences. 10.3390/ijms18051037. PubMed
3. Miyajima M,et al. 2017. Nat Immunol.. 10.1038/ni.3867. PubMed
4. Chmielewski M and Abken H 2017. Cell Rep.. 10.1016/j.celrep.2017.11.063. PubMed
5. Lin JR et al. 2018. eLife. 7 pii: e31657. PubMed
6. Stone S et al. 2018. Glia. 66(7):1331-1345 . PubMed
7. Huang L, et al. 2018. Exp Neurol. 300:41. PubMed
8. Simonneau M, et al. 2018. Oncotarget. 9:36457. PubMed
9. Pan RY, et al. 2019. Sci Adv. 5:eaau6328. PubMed
10. Lin M, et al. 2019. Mol Med Rep. 19:1622. PubMed
11. Rangan P et al. 2019. Cell reports. 26(10):2704-2719 . PubMed
12. Bochenek MA, et al. 2018. Nat Biomed Eng. 2:810. PubMed
13. Suzuki K, et al. 2019. Cell Mol Gastroenterol Hepatol. 8:119. PubMed
14. Laura C Burzynski et al. 2019. Immunity. 50(4):1033-1042 . PubMed
15. Garg BK, et al. 2019. PLoS One. 14:e0214942. PubMed
16. Farsakoglu Y et al. 2019. Cell reports. 26(9):2307-2315 . PubMed
17. Ooi JD, et al. 2019. Nat Commun. 10:3392. PubMed
18. He J, et al. 2019. J Clin Invest. 130. PubMed
19. Berry MR et al. 2017. Cell. 170(5):860-874 . PubMed
20. Hickman SE, et al. 2019. Methods Mol Biol. 2034:305. PubMed
21. Sparber F, et al. 2019. Cell Host Microbe. 25:389. PubMed
22. von Roemeling CA, et al. 2020. Nat Commun. 11:1508. PubMed
23. Silva HM, et al. 2019. J Exp Med. 216:786. PubMed
24. Chen C, et al. 2019. Cell Rep. 29:4200. PubMed
25. Cai B, et al. 2020. Cell Metabolism. 31(2):406-421.e7.. PubMed
26. Mirando AC, et al. 2020. Oncoimmunology. 9:1760685. PubMed
27. Gong Y, et al. 2020. J Neuroinflammation. 0.845833333. PubMed
28. Kim SH, et al. 2020. Neoplasia. 1.3375. PubMed
29. Wu X, et al. 2020. Front Immunol. 1.922222222. PubMed
30. Usui H, et al. 2007. Diabetes. 56:363. PubMed
31. Roychoudhury J, et al. 2010. Invest Ophthalmol Vis Sci. 51:3560. PubMed
32. Panka D, et al. 2013. Mol Cancer. 12:17. PubMed
33. Schwartz C, et al. 2014. J Immunol. 193:3590. PubMed
34. Onishi S, et al. 2015. PLoS One. 10:126564. PubMed
35. Venosa A, et al. 2015. Am J Physiol Lung Cell Mol Physiol. 309: L1487 - L1498. PubMed
36. Kar S, Colino J, Snapper C 2016. J Immunol. 196: 4204 - 4213. PubMed
37. Yamada K, et al. 2016. Cancer Res . 76: 4283 - 4292. PubMed
38. Hilpert C, et al. 2016. J Immunol. 197: 2780 - 2786. PubMed
39. Jiang H, et al. 2016. J Immunol. 197: 2880 - 2890. PubMed
40. Li X, et al. 2020. Aging Dis. 0.821527778. PubMed
41. Cheng Q, et al. 2020. Nat Nanotechnol. 0.842361111. PubMed
42. Rai MA, et al. 2020. Retrovirology. 17:35. PubMed
43. Barberio AE, et al. 2020. ACS Nano. 14:11238. PubMed
44. Perner C, et al. 2020. Immunity. 53(5):1063-1077.e7. PubMed
45. Uzhachenko RV, et al. 2021. Cell Reports. 35(1):108944. PubMed

RRID

AB_493545 (BioLegend Cat. No. 101219)
AB_389305 (BioLegend Cat. No. 101217)

Antigen Details

Structure

Integrin family, associates with integrin β₂ (CD18), 170 kD

Distribution

Granulocytes, monocytes/macrophages, dendritic cells, NK cells, subsets of T and B cells

Function

Adhesion, chemotaxis

Ligand/Receptor

ICAM-1 (CD54), ICAM-2 (CD102), ICAM-4 (CD242), iC3b, fibrinogen

Cell Type

B cells, Dendritic cells, Granulocytes, Macrophages, Monocytes, Neutrophils, NK cells, T cells, Tregs

Biology Area

Cell Adhesion, Cell Biology, Costimulatory Molecules, Immunology, Innate Immunity, Neuroscience, Neuroscience Cell Markers

Molecular Family

Adhesion Molecules, CD Molecules

Antigen References

1. Barclay A, et al. 1997. The Leukocyte Antigen FactsBook Academic Press.
2. Springer TA. 1994. Cell 76:301.
3. Coxon A, et al. 1996. Immunity 5:653.

Gene ID

16409 View all products for this Gene ID 3684 View all products for this Gene ID

UniProt

View information about CD11b on UniProt.org

Documentation

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

Reviews

Related Protocols

• Cell Surface Flow Cytometry Staining Protocol
• Ce3D™ Tissue Clearing Kit

Related Pages & Pathways

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.

Other Formats

Concentration & Expiration Lookup

Certificate of Analysis