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Purified 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_Purified_062707
Human peripheral blood lymphocytes stained with purified UCHL1, followed by anti-mouse IgGs FITC
  • A_UCHL1_Purified_062707
    Human peripheral blood lymphocytes stained with purified UCHL1, followed by anti-mouse IgGs FITC
  • B_UCHL1_PURE_CD45RO_Antibody_SB_111423
    SeqIF™ (sequential immunofluorescence) staining on COMET™ of Purified anti-CD45RO (clone UCHL1, yellow) on formalin-fixed paraffin-embedded human tonsil tissue at 2.5 µg/mL. Alexa Fluor™ Plus 647 Goat anti-Mouse IgG antibody (Lunaphore, Cat. No. DR647MS) was used as a secondary antibody. Nuclei were counterstained with DAPI (blue). Tissue underwent an all-in-one dewaxing and antigen retrieval preprocessing.
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304202 100 µg £35
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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
Phosphate-buffered solution, pH 7.2, containing 0.09% sodium azide.
Preparation
The antibody was purified by affinity chromatography.
Concentration
0.5 mg/ml
Storage & Handling
The antibody solution should be stored undiluted between 2°C and 8°C.
Application

FC - Quality tested
WB, IP, IHC-P - Reported in the literature, not verified in house
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 ≤ 2.0 µg per million cells in 100 µl volume. It is recommended that the reagent be titrated for optimal performance for each application.

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 applications, we have partnered with Lunaphore Technologies for demonstration of our antibodies on the COMET™. The COMET™ platform is an automated, end-to-end spatial biology solution developed for rapid and flexible multiplex tissue profiling. More information on the COMET™ and a complete list of our antibodies that have been demonstrated on the COMET™ 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. Rouers A, et al. 2021. Cell Rep Med. 2:100278. PubMed
  2. McIlwain DR, et al. 2021. Cell Host Microbe. 29:1828. PubMed
  3. Jaiswal A, et al. 2022. Cancer Cell. 40:524. PubMed
  4. Fischer JR, et al. 2023. Cell Rep Med. 4:100977. PubMed
  5. Friebel E, et al. 2020. Cell. 181(7):1626-1642.e20. PubMed
  6. Delgobo M, et al. 2021. Front Immunol. 12:584538. PubMed
  7. Kaufmann M, et al. 2021. Med. 2(3):296-312.e8. PubMed
  8. Liu F, et al. 2020. J Immunother Cancer. 8:. PubMed
  9. Ferrian S, et al. 2021. Cell Rep Med. 2:100419. PubMed
  10. Guo T, et al. 2018. J Immunol. 200:500. PubMed
  11. Cobb DA, et al. 2022. J Immunother Cancer. 10:. PubMed
  12. Cosgrove PR, et al. 2021. Pediatr Res. . PubMed
  13. Wu Y, et al. 2007. Infect Immun. 75:4357. PubMed
  14. Xu-Monette ZY, et al. 2019. Cancer Immunol Res. 7:644. PubMed
  15. Hazenberg MD, et al. 2019. Blood Adv. 2.659722222. PubMed
  16. Del Alcazar D, et al. 2019. Cell Rep. 28:3047. PubMed
  17. Gee MH, et al. 2018. Cell. 172:549. PubMed
  18. Mozaffarian N, et al. 2008. Rheumatology. 47:1335. PubMed
  19. Wu Y, et al. 2007. Infect Immun . 178:2802. PubMed
  20. Chng MHY, et al. 2020. Immunity. 51(6):1119-1135.e5.. PubMed
  21. Singh AK, et al. 2017. Front Immunol. 0.513888889. PubMed
  22. Sergio Gonzalez‐Duque et al. 2018. Cell metabolism. 28(6):946-960 . PubMed
  23. White M, et al. 2015. J Immunol. 195: 1858-1867. PubMed
  24. Mishra A, et al. 2021. Cell. 184(13):3394-3409.e20. PubMed
  25. NULL, et al. 2022. Cell. 185:916. PubMed
  26. Bender C, et al. 2020. Sci Adv. :6. PubMed
  27. Chang MH, et al. 2021. Cell Rep. 37:109902. PubMed
  28. Martin E, et al. 2020. JCI Insight. :5. PubMed
  29. Li Z, et al. 2020. J Clin Lab Anal. 34:e23155. PubMed
RRID
AB_314418 (BioLegend Cat. No. 304202)

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

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: 4    Revision Date: 11/14/2023

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

 

This product is supplied subject to the terms and conditions, including the limited license, located at www.biolegend.com/terms) ("Terms") and may be used only as provided in the Terms. Without limiting the foregoing, BioLegend products may not be used for any Commercial Purpose as defined in the Terms, resold in any form, used in manufacturing, or reverse engineered, sequenced, or otherwise studied or used to learn its design or composition without express written approval of BioLegend. Regardless of the information given in this document, user is solely responsible for determining any license requirements necessary for user’s intended use and assumes all risk and liability arising from use of the product. BioLegend is not responsible for patent infringement or any other risks or liabilities whatsoever resulting from the use of its products.

 

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Toll-Free Phone: 1-877-Bio-Legend (246-5343) Phone: (858) 768-5800 Fax: (877) 455-9587

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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