Biolegend, 304244, PE anti-human CD45RO Antibody, 100μg

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.
100μg
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/mLtest 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. UCHL1 signal intensity may be reduced in the presence of True-Stain Monocyte Blocker™ (Cat. No. 426102). Clone S19021B is not impacted by Monocyte Blocker and is recommended as an alternative to UCHL1.
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): Knapp W, et al. Eds. 1989. Leucocyte Typing IV. Oxford University Press. New York. (FC) Ishii T, et al. 2001. P. Natl. Acad. Sci. USA 98:12138. (WB) Ponsford M, et al. 2001. Clin. Exp. Immunol. 124:315. (IP) Yamada M, et al. 1996. Stroke 27:1155. (IHC) Sakkas LI, et al. 1998. Clin. Diagn. Lab. Immunol. 5:430. (IHC) Baba N, et al. 2010. Int. Immunol. 22:237. PubMed Thakral D, et al. 2008. J. Immunol. 180:7431. (FC) PubMed Weiss L, et al. 2010. P. Natl. Acad. Sci. USA 107:10632. PubMed Wu YY, et al. 2007. Infect. Immun. 75:4357. PubMed Mozaffarian N, et al. 2008. Rheumatology 47:1335. PubMed Roque S, et al. 2007. J. Immunol. 178:8028. PubMed Yoshino N, et al. 2000. Exp. Anim. (Tokyo) 49:97. (FC) Smith SH, et al. 1986. Immunology 58:63. (Immunogen) Peterson VM, et al. 2017. Nat. Biotechnol. 35:936. (PG)
Product Citations: Li M, et al. 2023. Front Immunol. 14:1087923. PubMed Si W, et al. 2023. iScience. 26:106529. PubMed Crawford MP, et al. 2023. PLoS One. 18:e0285166. PubMed Argüello RJ, et al. 2020. Cell Metab. 32:1063. PubMed Li M, et al. 2021. J Clin Invest. 131:. PubMed Lee YJ, et al. 2021. Front Immunol. 615369:12. PubMed Jarosch S, et al. 2022. STAR Protoc. 3:101374. PubMed Carisey AF, et al. 2018. Curr Biol. 28:489. PubMed Wang Z, et al. 2021. Cell Mol Immunol. 18:2188. PubMed Sutavani R, et al. 2013. J Immunol. 191:5895. PubMed Guo J, et al. 2022. Front Cell Dev Biol. 9:775599. PubMed Wu K, et al. 2021. Aging (Albany NY). 13:. PubMed Elston L, et al. 2020. Br J Haematol. 188:872. PubMed Iannetta M, et al. 2016. PLoS One. 11: 0160277. PubMed Narsale A, Moya R, Robertson H 2016. Data Brief. 8: 1348-51. PubMed Islas-Vazquez L, et al. 2020. Biology (Basel). 9:00. PubMed Evans RDR, et al. 2020. Nat Commun. 3.491666667. PubMed Murakami T, et al. 2018. Nat Commun. 9:2436. PubMed Wu D, et al. 2020. Biomark Res. 8:3. PubMed Jarosch S, et al. 2021. Cell Rep Methods. 1:100104. PubMed Godbersen C, et al. 2017. Mol Cancer Ther. 16:1335. PubMed Hirama T, et al. 2021. JCI Insight. 6:. PubMed Zhao B, et al. 2013. PLoS One. 8:77708. PubMed Zhou Z, et al. 2022. J Gastrointest Oncol. 13:732. PubMed Heckler M, et al. 2021. Cancer Discov. Online ahead of prin. PubMed Corrado M, et al. 2020. Cell Metab. 32:981. PubMed Li M, et al. 2020. Nat Commun. 4051:11. PubMed Shan L, et al. 2017. Immunity. 47:766. PubMed Chang MH, et al. 2021. Cell Rep. 37:109902. PubMed
RRID: AB_314421 (BioLegend Cat. No. 304205) AB_314422 (BioLegend Cat. No. 304206) AB_2564160 (BioLegend Cat. No. 304244)
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
UniProt: View information about CD45RO on UniProt.org
Clone: UCHL1
Regulatory Status: RUO
Workshop: IV N31
Other Names: CD45RO
Isotype: Mouse IgG2a, κ
Q: What type of PE do you use in your conjugates?
A: We use R-PE in our conjugates.
Q: If an antibody clone has been previously successfully used in IBEX in one fluorescent format, will other antibody formats work as well?
A: 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.
Q: Will antibodies my lab is already using for fluorescent or chromogenic IHC work in IBEX?
A: 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.
Q: Are other fluorophores compatible with IBEX?
A: 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.
Q: The same antibody works in one tissue type but not another. What is happening?
A: 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.
Q: How can I be sure the staining I’m seeing in my tissue is real?
A: 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.