Alexa Fluor® 488 anti-mouse CD45 Antibody

Product Details

Verified Reactivity

Mouse

Antibody Type

Monoclonal

Host Species

Rat

Immunogen

Mouse thymus or spleen

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

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. 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.

View full statement regarding label licenses

Excitation Laser

Blue Laser (488 nm)

Application Notes

Clone 30-F11 reacts with all isoforms and both CD45.1 and CD45.2 alloantigens of CD45.

Additional reported applications (for relevant formats) include: immunoprecipitation³, complement-dependent cytotoxicity^1,5, immunohistochemistry (acetone-fixed frozen sections, zinc-fixed paraffin-embedded sections and formalin-fixed paraffin-embedded sections)^4,6, Western blotting⁷, and spatial biology (IBEX)^10,11. The Ultra-LEAF™ purified antibody (Endotoxin < 0.01 EU/µg, Azide-Free, 0.2 µm filtered) is recommended for functional assays (Cat. No. 103163 and 103164).

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. Podd BS, et al. 2006. J. Immunol. 176:6532. (FC, CMCD) PubMed
2. Haynes NM, et al. 2007. J. Immunol. 179:5099. (FC)
3. Ledbetter JA, et al. 1979. Immunol. Rev. 47:63. (IP)
4. Simon DI, et al. 2000. J. Clin. Invest. 105:293. (IHC)
5. Seaman WE. 1983. J. Immunol. 130:1713. (CMCD)
6. Cornet A, et al. 2001. P. Natl. Acad. Sci. USA 98:13306. (IHC)
7. Tsuboi S and Fukuda M. 1998. J. Biol. Chem. 273:30680. (WB) PubMed
8. Liu F, et al. 2012. Blood. 119:3295. PubMed
9. Pelletier AN, et al. 2012. J. Immunol. 188:5561. PubMed
10. Radtke AJ, et al. 2020. Proc Natl Acad Sci U S A. 117:33455-65. (SB) PubMed
11. Radtke AJ, et al. 2022. Nat Protoc. 17:378-401. (SB) PubMed

Product Citations

1. Preda MB, et al. 2021. Cell Death Dis. 12:566. PubMed
2. Kuo PC, et al. 2021. Brain Commun. 3:fcab187. PubMed
3. Wu Y, et al. 2021. Immunity. 54:2595. PubMed
4. Wemlinger SM, et al. 2022. J Immunol. 208:1566. PubMed
5. Liu H, et al. 2022. Cell Rep Med. 3:100660. PubMed
6. Frederico B, et al. 2022. Dev Cell. 57:1957. PubMed
7. Ullah I, et al. 2023. Cell Rep Med. 4:100893. PubMed
8. Olivares-González L, et al. 2022. Antioxidants (Basel). 12: . PubMed
9. Gilman KE, et al. 2023. Front Immunol. 13:1045710. PubMed
10. Parida PK, et al. 2022. Cell Metab. 34:90. PubMed
11. Gomez-Salinero JM, et al. 2022. Cell Stem Cell. 29:593. PubMed
12. Luff SA, et al. 2022. Nat Cell Biol. 24:616. PubMed
13. Battis K, et al. 2022. J Neurosci. 42:7673. PubMed
14. Abdel-Haq R, et al. 2022. Elife. 11:. PubMed
15. Axelrod ML, et al. 2022. Nature. 611:818. PubMed
16. Gómez-Salinero JM, et al. 2022. Nat Cardiovasc Res. 1:882. PubMed
17. Fan Z, et al. 2023. Angiogenesis. . PubMed
18. Gilman KE, et al. 2023. Cancers (Basel). 15:. PubMed
19. Palakurthi B, et al. 2023. Nat Commun. 14:2109. PubMed
20. Fixsen BR, et al. 2023. Nat Immunol. . PubMed
21. Marko C, et al. 2014. Invest Ophthalmol Vis Sci . 55:291. PubMed
22. Giorgetti E, et al. 2020. Cell Reports. 29(6):1539-1554.e7.. PubMed
23. Kazakevych J, et al. 2020. Genome Biol. 21:64. PubMed
24. Sakai M, et al. 2020. Immunity. 51(4):655-670. PubMed
25. Schmidleithner L et al. 2019. Immunity. 50(5):1232-1248 . PubMed
26. Logan C, Bowen C, and Menko A 2017. Sci Rep. . 10.1038/s41598-017-16456-5. PubMed
27. Petursdottir D, et al. 2017. Front Immunol. . 10.3389/fimmu.2017.01699. PubMed
28. Nagai Y, et al. 2019. Front Immunol. 10:174. PubMed
29. Khandelwal P, et al. 2013. PLoS One. 8:64193. PubMed
30. Masschelein E, et al. 2020. Skelet Muscle. 10:21. PubMed
31. Martínez-Sabadell A, et al. 2022. STAR Protoc. 3:101712. PubMed
32. Rodriguez AB, et al. 2021. Cell Reports. 36(3):109422. PubMed
33. Bornes L, et al. 2021. Life Sci Alliance. 4:00. PubMed
34. Jenkins RW, et al. 2018. Cancer Discov. 8:196. PubMed
35. Dubey LK, et al. 2019. Cell Rep. 27:2442. PubMed
36. DeDreu J, et al. 2020. FASEB J. 34:9316. PubMed
37. Dos Santos Dias L, et al. 2021. PLoS Pathog. e1009324:17. PubMed
38. Hallett JM, et al. 2022. Cell Stem Cell. 29:355. PubMed
39. Mehta AK, et al. 2021. Nat Cancer. 2:66. PubMed
40. Yip HYK, et al. 2021. STAR Protocols. 2(3):100765. PubMed
41. Chen I, et al. 2016. Sci Rep. 6:27195. PubMed
42. Duarte J, et al. 2016. J Immunol. 197(12):4838-4847. PubMed
43. Jung Y, et al. 2021. Nat Commun. 12:3872. PubMed
44. Scarneo SA, et al. 2022. Sci Rep. 12:18091. PubMed
45. Gutknecht M, et al. 2017. Nat Commun. 10.1038/s41467-017-00488-6. PubMed
46. Ferrari G, et al. 2011. Invest Ophthalmol Vis Sci. 52:2532. PubMed
47. Mizbani A, et al. 2016. Development. 143: 4137 - 4148. PubMed
48. Burns JC, et al. 2020. eLife. 9:00. PubMed
49. Wara AK, et al. 2020. Cell Rep. 33:108550. PubMed
50. Wu H, et al. 2021. Cell Death Discov. 7:225. PubMed
51. Lustgarten Guahmich N, et al. 2020. Angiogenesis. 23:443. PubMed
52. Limbad C, et al. 2022. iScience. 25:103848. PubMed
53. Yip HYK, et al. 2020. Molecular Cell. 80(2):279-295.e8. PubMed
54. Liu X, et al. 2020. Cell Host Microbe. 28(5):683-698.e6. PubMed
55. Dobson HE, et al. 2020. Mucosal Immunol. 0.901388889. PubMed
56. Biton M et al. 2018. Cell. 175(5):1307-1320 . PubMed
57. Alam Z, et al. 2020. Cell Rep. 107825:31. PubMed
58. Shibad V, et al. 2021. Front Immunol. 12:722451. PubMed
59. Wüthrich M, et al. 2021. MBio. 12:e0201821. PubMed
60. Li D, et al. 2022. Commun Biol. 5:271. PubMed
61. Braun D, et al. 2013. J Biol Chem. 288:2689. PubMed
62. Pang M, et al. 2009. J Immunol. 182:7001. PubMed
63. Wu X, et al. 2018. Cell. 172:423. PubMed
64. von Roemeling CA, et al. 2020. Nat Commun. 11:1508. PubMed
65. Ding H, et al. 2016. Nat Commun. 7:11533. PubMed
66. Franklin DA, et al. 2020. JCI Insight. 5:. PubMed
67. Steele NG, et al. 2021. Clin Cancer Res. 27:2023. PubMed
68. Kazakevych J, et al. 2019. Sci Rep. 9:10410. PubMed
69. Peng W, et al. 2018. Mol Cell Biol. 38:e00427. PubMed
70. Kuo P, et al. 2016. J Am Heart Assoc. 5: 002610. PubMed
71. Markworth JF, et al. 2020. JCI Insight. 5:00. PubMed
72. Reed-Geaghan EG, et al. 2020. J Exp Med. 217:00:00. PubMed
73. Pantelidou C, et al. 2022. NPJ Breast Cancer. 8:102. PubMed
74. Lasch M, et al. 2020. Front Physiol. 11:576736. PubMed
75. DeDreu J, et al. 2022. FASEB J. 36:e21995. PubMed
76. Li Q, et al. 2020. Am J Pathol. 190:2453. PubMed
77. Lee EKS, et al. 2018. Cell Host Microbe. 23:121. PubMed
78. Wang Y, et al. 2019. Front Cell Infect Microbiol. 9:286. PubMed
79. Ye Y, et al. 2011. Am J Physiol Lung Cell Mol Physiol. 300:L216. PubMed
80. RL M, et al. 2015. Proc Natl Acad Sci U S A. 112:6506-6514. PubMed
81. Tian F, et al. 2016. Nat Commun. 7:13283. PubMed
82. Yang FM, et al. 2022. Front Immunol. 13:918241. PubMed
83. Lal JC, et al. 2021. Breast Cancer Res. 23:83. PubMed
84. Xiang W, et al. 2020. Signal Transduct Target Ther. 0.374305556. PubMed
85. Denk F, et al. 2016. Cell Rep. 15: 1771-1781. PubMed
86. Qi S, et al. 2020. Theranostics. 10:1814. PubMed

RRID

AB_493531 (BioLegend Cat. No. 103121)
AB_493531 (BioLegend Cat. No. 103122)

Antigen Details

Structure

Protein tyrosine phosphatase (PTP) family, 180-240 kD

Distribution

All hematopoietic cells except mature erythrocytes and platelets

Function

Phosphatase, T and B cell activation

Ligand/Receptor

Galectin-1, CD2, CD3, CD4, TCR, CD22, Thy-1

Cell Type

B cells, Dendritic cells, Mesenchymal Stem Cells, Tregs

Biology Area

Cell Biology, Immunology, Inhibitory Molecules, Innate Immunity, Neuroscience, Neuroscience Cell Markers, Stem Cells

Molecular Family

CD Molecules

Antigen References

1. Barclay A, et al. 1997. The Leukocyte Antigen FactsBook Academic Press.
2. Trowbridge IS, et al. 1993. Annu. Rev. Immunol. 12:85.
3. Kishihara K, et al. 1993. Cell 74:143.
4. Pulido R, et al. 1988. J. Immunol. 140:3851.

Gene ID

19264 View all products for this Gene ID

UniProt

View information about CD45 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

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