- RA3-6B2 (See other available formats)
- Regulatory Status
- Other Names
- Rat IgG2a, κ
- Ave. Rating
- Submit a Review
- Product Citations
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CD45R, also known as B220, is an isoform of CD45. It is a member of the protein tyrosine phosphatase (PTP) family with a molecular weight of approximately 180-240 kD. CD45R is expressed on B cells (at all developmental stages from pro-B cells through mature B cells), activated B cells, and subsets of T and NK cells. CD45R (B220) is also expressed on a subset of abnormal T cells involved in the pathogenesis of systemic autoimmunity in MRL-Faslpr and MRL-Fasgld mice. It plays a critical role in TCR and BCR signaling. The primary ligands for CD45 are galectin-1, CD2, CD3, and CD4. CD45R is commonly used as a pan-B cell marker; however, CD19 may be more appropriate for B cell specificity.Product Details
- Verified Reactivity
- Mouse, Human
- Reported Reactivity
- Antibody Type
- Host Species
- Abelson murine leukemia virus-induced pre-B tumor cells
- Phosphate-buffered solution, pH 7.2, containing 0.09% sodium azide.
- The antibody was purified by affinity chromatography and conjugated with Alexa Fluor® 647 under optimal conditions.
- 0.5 mg/mL
- 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.
FC - Quality tested
IHC-F, 3D IHC - Verified
SB - Reported in the literature, not verified in house
- 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 ≤ 0.25 µg per million cells in 100 µL volume. For immunohistochemistry on frozen tissue sections, a concentration range of 2.5 - 5.0 µg/mL is suggested. For immunofluorescence microscopy, a concentration range of 1.25 - 10 µg/mL is recommended. It is recommended that the reagent be titrated for optimal performance for each application.
* Alexa Fluor® 647 has a maximum emission of 668 nm when it is excited at 633nm / 635nm.
Alexa Fluor® and Pacific Blue™ are trademarks of Life Technologies Corporation.
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- Excitation Laser
Red Laser (633 nm)
- Application Notes
Clone RA3-6B2 has been described to react with an epitope on the extracellular domain of the transmembrane CD45 glycoprotein which is dependent upon the expression of exon A and specific carbohydrate residues. Additional reported applications (for the relevant formats) include: immunoprecipitation1, in vitro and in vivo modulation of B cell responses2-4, immunohistochemistry of acetone-fixed frozen sections and formalin-fixed paraffin-embedded sections5,6, and spatial biology (IBEX)14,15.
- Additional Product Notes
Iterative Bleaching Extended multi-pleXity (IBEX) is a fluorescent imaging technique capable of highly-multiplexed spatial analysis. The method relies on cyclical bleaching of panels of fluorescent antibodies in order to image and analyze many markers over multiple cycles of staining, imaging, and, bleaching. It is a community-developed open-access method developed by the Center for Advanced Tissue Imaging (CAT-I) in the National Institute of Allergy and Infectious Diseases (NIAID, NIH).
(PubMed link indicates BioLegend citation)
- Coffman RL. 1982. Immunol. Rev. 69:5. (IP)
- George A, et al. 1994. J. Immunol. 152:1014. (Activ)
- Asensi V, et al. 1989. Immunology 68:204. (Activ)
- Domiati-Saad R, et al. 1993. J. Immunol. 151:5936. (Activ)
- Hata H, et al. 2004. J. Clin. Invest. 114:582. (IHC)
- Monteith CE, et al. 1996. Can. J. Vet. Res. 60:193. (IHC)
- Shih FF, et al. 2006. J. Immunol. 176:3438. (FC)
- Chang C L-T, et al. 2007. J. Immunol. 178:6984.
- Fazilleau N, et al. 2007. Nature Immunol. 8:753.
- Lang GL, et al. 2008. Blood 111:2158. PubMed
- Charles N, et al. 2010. Nat. Med. 16:701. (FC) PubMed
- del Rio ML, et al. 2011. Transpl. Int. 24:501. (FC) PubMed
- Murakami R, et al. 2013. PLoS One. 8:73270. PubMed
- Radtke AJ, et al. 2020. Proc Natl Acad Sci U S A. 117:33455-65. (SB) PubMed
- Radtke AJ, et al. 2022. Nat Protoc. 17:378-401. (SB) PubMed
- Product Citations
AB_492875 (BioLegend Cat. No. 103229)
AB_389330 (BioLegend Cat. No. 103226)
- Protein tyrosine phosphatase (PTP) family, 180-240 kD
B cells, T cell subset, NK cell subset
- Phosphatase, T and B cell activation
- Galectin-1, CD2, CD3, CD4
- Cell Type
- B cells, NK cells, T cells
- Biology Area
- Cell Biology, Immunology, Inhibitory Molecules, Neuroscience, Neuroscience Cell Markers
- Molecular Family
- CD Molecules
- Antigen References
1. Barclay A, et al. 1997. The Leukocyte Antigen FactsBook Academic Press.
2. Trowbridge IS, et al. 1993. Annu. Rev. Immunol. 12:85.
3. Kishihara K, et al. 1993. Cell 74:143.
4. Pulido R, et al. 1988. J. Immunol. 140:3851.
- Gene ID
- 19264 View all products for this Gene ID 5788 View all products for this Gene ID
- View information about CD45R on UniProt.org
- 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.
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Your actual data may vary due to variations in samples, target cells, instruments and their settings, staining conditions, and other factors.
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Alexa Fluor® 594 anti-mouse/human CD45R/B220
APC anti-mouse/human CD45R/B220
Biotin anti-mouse/human CD45R/B220
FITC anti-mouse/human CD45R/B220
PE anti-mouse/human CD45R/B220
PE/Cyanine5 anti-mouse/human CD45R/B220
Purified anti-mouse/human CD45R/B220
PE/Cyanine7 anti-mouse/human CD45R/B220
APC/Cyanine7 anti-mouse/human CD45R/B220
Alexa Fluor® 488 anti-mouse/human CD45R/B220
Alexa Fluor® 647 anti-mouse/human CD45R/B220
Pacific Blue™ anti-mouse/human CD45R/B220
Alexa Fluor® 700 anti-mouse/human CD45R/B220
PerCP anti-mouse/human CD45R/B220
PerCP/Cyanine5.5 anti-mouse/human CD45R/B220
Brilliant Violet 421™ anti-mouse/human CD45R/B220
Brilliant Violet 570™ anti-mouse/human CD45R/B220
Brilliant Violet 650™ anti-mouse/human CD45R/B220
Brilliant Violet 605™ anti-mouse/human CD45R/B220
Brilliant Violet 785™ anti-mouse/human CD45R/B220
Brilliant Violet 510™ anti-mouse/human CD45R/B220
Purified anti-mouse/human CD45R/B220 (Maxpar® Ready)
Brilliant Violet 711™ anti-mouse/human CD45R/B220
PE/Dazzle™ 594 anti-mouse/human CD45R/B220
APC/Fire™ 750 anti-mouse/human CD45R/B220
Brilliant Violet 750™ anti-mouse/human CD45R/B220
TotalSeq™-A0103 anti-mouse/human CD45R/B220
Spark Blue™ 550 anti-mouse/human CD45R/B220
Spark NIR™ 685 anti-mouse/human CD45R/B220
TotalSeq™-B0103 anti-mouse/human CD45R/B220
Ultra-LEAF™ Purified anti-mouse/human CD45R/B220
TotalSeq™-C0103 anti-mouse/human CD45R/B220
PE/Fire™ 640 anti-mouse/human CD45R/B220
APC/Fire™ 810 anti-mouse/human CD45R/B220
PE/Fire™ 700 anti-mouse/human CD45R/B220
Spark Violet™ 538 anti-mouse/human CD45R/B220
Spark YG™ 581 anti-mouse/human CD45R/B220
Spark YG™ 570 anti-mouse/human CD45R/B220
PE/Fire™ 810 anti-mouse/human CD45R/B220
Spark Blue™ 574 anti-mouse/human CD45R/B220 Antibody
Spark Violet™ 423 anti-mouse/human CD45R/B220 Antibody
Spark Red™ 718 anti-mouse/human CD45R/B220