PE anti-human CD45RO Antibody

Pricing & Availability
Clone
UCHL1 (See other available formats)
Regulatory Status
RUO
Workshop
IV N31
Other Names
CD45RO
Isotype
Mouse IgG2a, κ
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Product Citations
publications
a-UCHL1
Human peripheral blood lymphocytes stained with UCHL1 PE
  • a-UCHL1
    Human peripheral blood lymphocytes stained with UCHL1 PE
  • b-UCHL1_PE_072909.jpg
    Human peripheral blood lymphocytes stained with UCHL1 PE and CD45RA (HI100) Alexa Fluor® 647
  • c-UCHL1_PE_CD45RO_Antibody_3_101024
    Multiplexed IHC staining of PE anti-CD45RO (clone UCHL1) on formalin-fixed paraffin-embedded human tonsil tissue, validated for use on the Cellscape™. The tissue was iteratively stained with PE anti-CD45RO (clone UCHL1, green) and Alexa Fluor® 488 anti-CD3 (red) for one hour at room temperature. Nuclei were counterstained with DAPI. Images were captured with a 20X objective. Scale bar: 50 µm
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304205 25 tests 40€
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304206 100 tests 88€
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304244 100 µg 94€
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Description

CD45RO is a 180 kD single chain membrane glycoprotein. It is a splice variant of tyrosine phosphatase CD45, lacking the A, B, and C determinants. The CD45RO isoform is expressed on activated and memory T cells, some B cell subsets, activated monocytes/macrophages, and granulocytes. CD45RO enhances both T cell receptor and B cell receptor signaling mediated activation. CD45 and its isoforms non-covalently associate with lymphocyte phosphatase-associated phosphoprotein (LPAP) on T and B lymphocytes. CD45 has been reported to be associated with several other cell surface antigens including CD1, CD2, CD3, and CD4. CD45 has also been reported to bind galectin-1 and CD22. CD45 isoform expression can change in response to cytokines.

Product Details
Technical Data Sheet (pdf)

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
  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. Lee YJ, et al. 2021. Front Immunol. 615369:12. PubMed
  7. Jarosch S, et al. 2022. STAR Protoc. 3:101374. 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. Sutavani R, et al. 2013. J Immunol. 191:5895. PubMed
  11. Guo J, et al. 2022. Front Cell Dev Biol. 9:775599. PubMed
  12. Wu K, et al. 2021. Aging (Albany NY). 13:. 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. Zhao B, et al. 2013. PLoS One. 8:77708. PubMed
  24. Zhou Z, et al. 2022. J Gastrointest Oncol. 13:732. 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
UniProt
View information about CD45RO on UniProt.org

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

View All CD45RO Reagents Request Custom Conjugation
Description Clone Applications
APC anti-human CD45RO UCHL1 FC
FITC anti-human CD45RO UCHL1 FC
PE anti-human CD45RO UCHL1 FC,SB
PE/Cyanine5 anti-human CD45RO UCHL1 FC
Purified anti-human CD45RO UCHL1 FC,WB,IP,IHC-P,SB
Alexa Fluor® 488 anti-human CD45RO UCHL1 FC,IHC-P
Pacific Blue™ anti-human CD45RO UCHL1 FC
Alexa Fluor® 700 anti-human CD45RO UCHL1 FC
Biotin anti-human CD45RO UCHL1 FC
Brilliant Violet 421™ anti-human CD45RO UCHL1 FC,IHC-P
PerCP/Cyanine5.5 anti-human CD45RO UCHL1 FC
Brilliant Violet 570™ anti-human CD45RO UCHL1 FC
Brilliant Violet 605™ anti-human CD45RO UCHL1 FC
APC/Cyanine7 anti-human CD45RO UCHL1 FC
PE/Cyanine7 anti-human CD45RO UCHL1 FC
Brilliant Violet 650™ anti-human CD45RO UCHL1 FC
Brilliant Violet 711™ anti-human CD45RO UCHL1 FC
Brilliant Violet 785™ anti-human CD45RO UCHL1 FC
Alexa Fluor® 594 anti-human CD45RO UCHL1 IHC-P
Purified anti-human CD45RO (Maxpar® Ready) UCHL1 FC,CyTOF®
Brilliant Violet 510™ anti-human CD45RO UCHL1 FC
PE/Dazzle™ 594 anti-human CD45RO UCHL1 FC
APC/Fire™ 750 anti-human CD45RO UCHL1 FC
PerCP anti-human CD45RO UCHL1 FC
APC anti-human CD45RO UCHL1 FC
TotalSeq™-A0087 anti-human CD45RO UCHL1 PG
TotalSeq™-B0087 anti-human CD45RO UCHL1 PG
TotalSeq™-C0087 anti-human CD45RO UCHL1 PG
Brilliant Violet 750™ anti-human CD45RO UCHL1 FC
PE/Fire™ 640 anti-human CD45RO UCHL1 FC
TotalSeq™-D0087 anti-human CD45RO UCHL1 PG
Spark Violet™ 423 anti-human CD45RO UCHL1 FC
GMP APC anti-human CD45RO UCHL1 FC
PE anti-human CD45RO UCHL1 FC
TotalSeq™-Bn1369 anti-human CD45RO UCHL1 SB
GMP PE anti-human CD45RO UCHL1 FC
Spark UV™ 387 anti-human CD45RO UCHL1 FC
APC/Fire™ 810 anti-human CD45RO UCHL1 FC
Go To Top Version: 2    Revision Date: 10-10-2024

For Research Use Only. Not for diagnostic or therapeutic use.

 

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This data display is provided for general comparisons between formats.
Your actual data may vary due to variations in samples, target cells, instruments and their settings, staining conditions, and other factors.
If you need assistance with selecting the best format contact our expert technical support team.

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