PE anti-human CD45RO Antibody

Product Details

Verified Reactivity

Human

Reported Reactivity

Chimpanzee, Cynomolgus, Common Marmoset

Antibody Type

Monoclonal

Host Species

Mouse

Immunogen

IL-2 dependent T cell line, CA1

Formulation

µg size: Phosphate-buffered solution, pH 7.2, containing 0.09% sodium azide.
test sizes: Phosphate-buffered solution, pH 7.2, containing 0.09% sodium azide and BSA (origin USA).

Preparation

The antibody was purified by affinity chromatography, and conjugated with PE under optimal conditions.

Concentration

µg sizes: 0.2 mg/mL
test sizes: lot-specific (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, and protected from prolonged exposure to light. Do not freeze.

Application

FC - Quality tested
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 using the µg size, the suggested use of this reagent is ≤0.2 µg per million cells in 100 µl volume. It is recommended that the reagent be titrated for optimal performance for each application. For flow cytometric staining using the test sizes, the suggested use of this reagent is 5 µl per million cells in 100 µl staining volume or 5 µl per 100 µl of whole blood.

Excitation Laser

Blue Laser (488 nm)
Green Laser (532 nm)/Yellow-Green Laser (561 nm)

Application Notes

The UCHL1 antibody is commonly used in combination with antibodies against CD45RA to discern memory and naïve T cells. Additional reported applications (for the relevant formats) include: immunohistochemical staining of acetone-fixed frozen tissue sections⁵ and formalin-fixed paraffin-embedded tissue sections⁴, Western blotting², and immunoprecipitation³.

Additional Product Notes

For the use of this antibody in spatial biology (SB), we have partnered with Bruker Spatial Biology Biosciences for demonstration of this antibody on their next-generation ChipCytometry instrument called the CellScape™. The CellScape platform is an end-to-end solution for highly multiplexed spatial omics. Combining an advanced, purpose-built imaging system with easy-to-use fluidics for walk-away automation, the CellScape system will accelerate your exploration into the rapidly evolving field of spatial biology. More information on the the Bruker Spatial Biology CellScape and a complete list of our antibodies that have been demonstrated on the instrument can be found here.

Application References

(PubMed link indicates BioLegend citation)

1. Knapp W, et al. Eds. 1989. Leucocyte Typing IV. Oxford University Press. New York. (FC)
2. Ishii T, et al. 2001. P. Natl. Acad. Sci. USA 98:12138. (WB)
3. Ponsford M, et al. 2001. Clin. Exp. Immunol. 124:315. (IP)
4. Yamada M, et al. 1996. Stroke 27:1155. (IHC)
5. Sakkas LI, et al. 1998. Clin. Diagn. Lab. Immunol. 5:430. (IHC)
6. Baba N, et al. 2010. Int. Immunol. 22:237. PubMed
7. Thakral D, et al. 2008. J. Immunol. 180:7431. (FC) PubMed
8. Weiss L, et al. 2010. P. Natl. Acad. Sci. USA 107:10632. PubMed
9. Wu YY, et al. 2007. Infect. Immun. 75:4357. PubMed
10. Mozaffarian N, et al. 2008. Rheumatology 47:1335. PubMed
11. Roque S, et al. 2007. J. Immunol. 178:8028. PubMed
12. Yoshino N, et al. 2000. Exp. Anim. (Tokyo) 49:97. (FC)
13. Smith SH, et al. 1986. Immunology 58:63. (Immunogen)
14. Peterson VM, et al. 2017. Nat. Biotechnol. 35:936. (PG)

Product Citations

1. Argüello RJ, et al. 2020. Cell Metab. 32:1063. PubMed
2. Li M, et al. 2021. J Clin Invest. 131:. PubMed
3. Li M, et al. 2023. Front Immunol. 14:1087923. PubMed
4. Si W, et al. 2023. iScience. 26:106529. PubMed
5. Crawford MP, et al. 2023. PLoS One. 18:e0285166. PubMed
6. Jarosch S, et al. 2022. STAR Protoc. 3:101374. PubMed
7. Lee YJ, et al. 2021. Front Immunol. 615369:12. PubMed
8. Wang Z, et al. 2021. Cell Mol Immunol. 18:2188. PubMed
9. Carisey AF, et al. 2018. Curr Biol. 28:489. PubMed
10. Guo J, et al. 2022. Front Cell Dev Biol. 9:775599. PubMed
11. Wu K, et al. 2021. Aging (Albany NY). 13:. PubMed
12. Sutavani R, et al. 2013. J Immunol. 191:5895. PubMed
13. Elston L, et al. 2020. Br J Haematol. 188:872. PubMed
14. Iannetta M, et al. 2016. PLoS One. 11: 0160277. PubMed
15. Narsale A, Moya R, Robertson H 2016. Data Brief. 8: 1348-51. PubMed
16. Islas-Vazquez L, et al. 2020. Biology (Basel). 9:00. PubMed
17. Evans RDR, et al. 2020. Nat Commun. 3.491666667. PubMed
18. Murakami T, et al. 2018. Nat Commun. 9:2436. PubMed
19. Wu D, et al. 2020. Biomark Res. 8:3. PubMed
20. Jarosch S, et al. 2021. Cell Rep Methods. 1:100104. PubMed
21. Godbersen C, et al. 2017. Mol Cancer Ther. 16:1335. PubMed
22. Hirama T, et al. 2021. JCI Insight. 6:. PubMed
23. Zhou Z, et al. 2022. J Gastrointest Oncol. 13:732. PubMed
24. Zhao B, et al. 2013. PLoS One. 8:77708. PubMed
25. Heckler M, et al. 2021. Cancer Discov. Online ahead of prin. PubMed
26. Corrado M, et al. 2020. Cell Metab. 32:981. PubMed
27. Li M, et al. 2020. Nat Commun. 4051:11. PubMed
28. Chang MH, et al. 2021. Cell Rep. 37:109902. PubMed
29. Shan L, et al. 2017. Immunity. 47:766. PubMed

RRID

AB_2564160 (BioLegend Cat. No. 304205)
AB_2564160 (BioLegend Cat. No. 304206)
AB_2564160 (BioLegend Cat. No. 304244)

Antigen Details

Structure

Tyrosine phosphatases, type I transmembrane, 180 kD (isoform of CD45 containing none of the A, B, or C determinants)

Distribution

Activated and memory T cells, B cell subsets, monocytes, macrophages, granulocytes

Function

Enhances TCR and BCR signaling

Ligand/Receptor

CD22

Cell Type

B cells, Granulocytes, Macrophages, Mesenchymal Stem Cells, Monocytes, T cells, Tregs

Biology Area

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

Molecular Family

CD Molecules

Antigen References

1. Thomas M. 1989. Annu. Rev. Immunol. 7:339.
2. Trowbridge I, et al. 1994. Annu. Rev. Immunol. 12:85.

Gene ID

5788 View all products for this Gene ID

Specificity (DOES NOT SHOW ON TDS):

CD45RO

Specificity Alt (DOES NOT SHOW ON TDS):

CD45RO

App Abbreviation (DOES NOT SHOW ON TDS):

FC,SB

UniProt

View information about CD45RO 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

What type of PE do you use in your conjugates?

We use R-PE in our conjugates.

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