Alexa Fluor® 647 anti-mouse/human CD11b Antibody

Product Details

Verified Reactivity

Mouse, Human, Cynomolgus, Rhesus

Reported Reactivity

Chimpanzee, Baboon, Rabbit

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® 647 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
3D IHC - Verified
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 ≤ 0.25 µg per 10⁶ cells in 100 µl volume. 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® 647 has a maximum emission of 668 nm when it is excited at 633nm / 635nm.

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

View full statement regarding label licenses

Excitation Laser

Red Laser (633 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

This product has been verified for IHC-F (Immunohistochemistry - frozen tissue sections) and IHC-P (Immunohistochemistry - formalin-fixed paraffin-embedded tissues) on the NanoString GeoMx® Digital Spatial Profiler. The GeoMx® enables researchers to perform spatial analysis of protein and RNA targets in FFPE and fresh frozen human and mouse samples. For more information about our spatial biology products and the GeoMx® platform, please visit our spatial biology page.

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. Alam A, et al. 2022. Cancer Cell. 40:153. PubMed
2. Replogle JM, et al. 2022. Cell. 185:2559. PubMed
3. Sweet R, et al. 2017. J Immunol. 10.4049/jimmunol.1600861. PubMed
4. Vaitaitis GM, et al. 2023. J Biol Chem. 299:104625. PubMed
5. Kang YA, et al. 2023. J Exp Med. 220:. PubMed
6. Lütge M, et al. 2023. Nat Immunol. . PubMed
7. Ortiz G, et al. 2020. Front Immunol. 11:1713. PubMed
8. Jackson A, et al. 2014. J Leukoc Biol. 92:609. PubMed
9. Abels ER et al. 2019. Cell Rep. 28(12):3105-3119 . PubMed
10. Miller EB, et al. 2019. Proc Natl Acad Sci U S A. 116:16603. PubMed
11. Chen G, et al. 2017. Biosens Bioelectron. 10.1016/j.bios.2016.10.015. PubMed
12. Farsakoglu Y et al. 2019. Cell reports. 26(9):2307-2315 . PubMed
13. Karlen SJ, et al. 2018. J Neuroinflammation. 15:344. PubMed
14. Pham THM, et al. 2020. Cell Host & Microbe. 27(1):54-67.e5.. PubMed
15. Miller EB, et al. 2021. J Neuroinflammation. 18:235. PubMed
16. Salei N, et al. 2020. J Am Soc Nephrol. 31:257. PubMed
17. Feng Y, et al. 2018. Kidney Dis (Basel). 4:95. PubMed
18. Davidson S, et al. 2020. Cell Reports. 31(7):107628. PubMed
19. Socodato R, et al. 2020. Sci Signal. 13: . PubMed
20. Singh A, et al. 2020. Mol Oncol. 1.901388889. PubMed
21. Wongchana W, et al. 2015. J Immunol. 195: 5337 - 5346. PubMed
22. Pronin A, et al. 2019. Front Mol Neurosci. 12:36. PubMed
23. Baxter PS, et al. 2021. Cell Rep. 34:108882. PubMed
24. Pierce H, et al. 2017. Cell Stem Cell. 1.283333333. PubMed
25. Pinho S et al. 2018. Developmental cell. 44(5):634-641 . PubMed
26. Philip E Boulais et al. 2018. Immunity. 49(4):627-639 . PubMed
27. Balzano M et al. 2019. Cell reports. 26(12):3257-3271 . PubMed
28. Szeifert V, et al. 2021. Front Immunol. 12:671995. PubMed
29. Wei Q, et al. 2020. Dev Cell. 53:503. PubMed
30. Nowak W, et al. 2020. EBioMedicine. 50:290-305.. PubMed
31. Cunha LD et al. 2018. Cell. 175(2):429-441 . PubMed
32. Richardson ET, et al. 2015. PLoS One. 10: 1371. PubMed
33. Laban H, et al. 2018. J Cell Biol. 217:1503. PubMed
34. Chen ST et al. 2019. Cell host & microbe. 25(4):602-616 . PubMed
35. Canedo T, et al. 2021. Neuropsychopharmacology. 46:2358. PubMed
36. Richardson E, et al. 2015. Infect Immun . 83: 2242-2254. PubMed
37. Sanders K, et al. 2015. Cancer Immunol Res. 3: 891-901. PubMed
38. Anderson DA, et al. 2022. J Exp Med. 219:. PubMed
39. Mesa-Nuñez C, et al. 2022. Mol Ther Methods Clin Dev. 26:459. PubMed
40. Chen W, et al. 2016. Nat Commun. 7: 11302. PubMed
41. Bade RM, et al. 2021. Molecular Oncology. . PubMed
42. Halder LD, et al. 2020. Nat Commun. 2.077083333. PubMed
43. Catarinella M, et al. 2016. EMBO Mol Med. 8: 155 - 170. PubMed
44. Oh B, et al. 2017. Plast Reconstr Surg Glob Open. 5:e1595. PubMed
45. Silva HM, et al. 2019. J Exp Med. 216:786. PubMed
46. Cruz F, et al. 2016. Stem Cells Trans Med. 5: 488-499. PubMed
47. Alberts A, et al. 2020. Front Immunol. 11:596103. PubMed
48. Kim SI, et al. 2020. Molecular Cancer Therapeutics. 20(1):173-182. PubMed
49. Li Z et al. 2018. Immunity. 49(4):640-653 . PubMed
50. Wu L, et al. 2021. Cell Death Dis. 12:1064. PubMed
51. Rasmussen J, et al. 2006. Infect Immun. 74:6590. PubMed
52. Maas SLN, et al. 2020. J Neuroinflammation. 17:120. PubMed

RRID

AB_389327 (BioLegend Cat. No. 101220)
AB_389327 (BioLegend Cat. No. 101218)

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
• Material 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

Certificate of Analysis