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TotalSeq™-Bn1369 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
Barcode Sequence
TATAGTTCACGCCTC
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304269 10 µg 369 CHF
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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 and EDTA
Preparation
The antibody was purified by chromatography and conjugated with TotalSeq™-Bn oligomer under optimal conditions.
Concentration
0.5 mg/mL
Storage & Handling
The antibody solution should be stored undiluted between 2°C and 8°C. Do not freeze.
Application

SB - Quality tested
Recommended Usage

Each lot of this antibody is quality control tested by immunofluorescent staining in formalin-fixed, paraffin-embedded (FFPE) lymphoid tissue, and the oligomer sequence is confirmed by sequencing. TotalSeq™-Bn antibodies are compatible with the 10x Visium CytAssist Gene and Protein Expression Assay.

To maximize performance, it is strongly recommended that the reagent be titrated for each application, and that you centrifuge the antibody dilution at 14,000xg at 2 − 8°C for 10 minutes before use. Carefully pipette out the liquid avoiding the bottom of the tube when handling. To determine and optimize dilutions for the addition of Totalseq™-Bn antibodies into pre-designed antibody panels, refer to 10x Genomics Custom Add-on Antibody Optimization guide.


Buyer is solely responsible for determining whether Buyer has all intellectual property rights that are necessary for Buyer's intended uses of the BioLegend TotalSeq™ products. For example, for any technology platform Buyer uses with TotalSeq™, it is Buyer's sole responsibility to determine whether it has all necessary third party intellectual property rights to use that platform and TotalSeq™ with that platform.
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

TotalSeq™-Bn reagents are designed to profile protein levels following an optimized protocol in spatial transcriptomics. Compatible spatial biology devices (e.g. Imaging System, 10x Genomics Visium Spatial CytAssist Gene and Protein Expression instruments and reagents) and sequencer (e.g. Illumina analyzers) are required. TotalSeq™-B reagents are not compatible with the 10x Genomics Visium system. The complete barcode sequence may be provided upon request. Please contact technical support for more information, or visit TotalSeq™-Bn Reagents for 10x Genomics Visium CytAssist Gene and Protein Assay.

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)
RRID
AB_3083169 (BioLegend Cat. No. 304269)

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
PerCP/Fire™ 806 anti-human CD45RO Antibody UCHL1 FC
PerCP/Fire™ 780 anti-human CD45RO Antibody UCHL1 FC
Go To Top Version: 1    Revision Date: 08.24.2023

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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