Alexa Fluor® 594 anti-mouse CD45.2 Antibody

Pricing & Availability
Clone
104 (See other available formats)
Regulatory Status
RUO
Other Names
Ly-5.2, LCA
Isotype
Mouse (SJL) IgG2a, κ
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Product Citations
publications
104_A594_CD45-2_Antibody_1_111419
BALB/C mouse frozen spleen section was fixed with 4% paraformaldehyde (PFA) for ten minutes at room temperature and blocked with 5% rat serum + FBS for one hour at room temperature. Then the section was stained with 10 µg/ml of CD45.2 (clone 104) Alexa Fluor® 594 (red), 5 µg/ml of CD3 (clone 17A2) Alexa Fluor® 647 (green), and 5 µg/ml of B220 (clone RA3-6B2) Alexa Fluor® 488 (blue) at room temperature for two hours. The image was captured with a 10X objective.
  • 104_A594_CD45-2_Antibody_1_111419
    BALB/C mouse frozen spleen section was fixed with 4% paraformaldehyde (PFA) for ten minutes at room temperature and blocked with 5% rat serum + FBS for one hour at room temperature. Then the section was stained with 10 µg/ml of CD45.2 (clone 104) Alexa Fluor® 594 (red), 5 µg/ml of CD3 (clone 17A2) Alexa Fluor® 647 (green), and 5 µg/ml of B220 (clone RA3-6B2) Alexa Fluor® 488 (blue) at room temperature for two hours. The image was captured with a 10X objective.
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109850 100 µg 179€
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Description

CD45.2 is an alloantigen of CD45, expressed by Ly5.2 bearing mouse strains (e.g., A, AKR, BALB/c, CBA/Ca, CBA/J, C3H/He, C57BL, C57BR, C57L, C58, DBA/1, DBA/2, NZB, SWR, 129). CD45, a member of the protein tyrosine phosphatase (PTP) family, is a 180-240 kD glycoprotein expressed on all hematopoietic cells except mature erythrocytes and platelets. There are multiple isoforms in the mouse that play key roles in TCR and BCR signal transduction. These isoforms are very specific to the activation and maturation states of the cell as well as specific cell type. The primary ligands for CD45 are galectin-1, CD2, CD3, CD4, TCR, CD22, and Thy-1.

Product Details
Technical Data Sheet (pdf)

Product Details

Verified Reactivity
Mouse
Antibody Type
Monoclonal
Host Species
Mouse
Immunogen
B10.S mouse thymocytes and splenocytes
Formulation
Phosphate-buffered solution, pH 7.2, containing 0.09% sodium azide.
Preparation
The antibody was purified by affinity chromatography and conjugated with Alexa Fluor® 594 under optimal conditions.
Concentration
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.
Application

IHC-F - Quality tested
SB - Community verified

Recommended Usage

Each lot of this antibody is quality control tested by immunohistochemical staining on frozen tissue sections. For immunohistochemistry, a concentration range of 5.0 - 10 μg/ml is suggested. It is recommended that the reagent be titrated for optimal performance for each application.

* Alexa Fluor® 594 has an excitation maximum of 590 nm, and a maximum emission of 617 nm.


Alexa Fluor® and Pacific Blue™ are trademarks of Life Technologies Corporation.

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Application Notes

The 104 antibody does not react with mouse cells expressing the CD45.1 alloantigen. Additional reported applications (for the relevant formats) include: immunoprecipitation4, in vivo and in vitro blocking of B cell responses1,2, and immunohistochemical staining of acetone-fixed frozen sections3

Additional Product Notes

This product has been verified for IHC-F (Immunohistochemistry - frozen tissue sections) on the NanoString GeoMx® Digital Spatial Profiler. The GeoMx® enables researchers to perform spatial analysis of protein and RNA targets in FFPE and fresh frozen human and mouse samples. For more information about our spatial biology products and the GeoMx® platform, please visit our spatial biology page.

Application References

(PubMed link indicates BioLegend citation)
  1. Yakura H, et al. 1983. J. Exp. Med. 157:1077. (Block)
  2. Yakura H, et al. 1986. J. Immunol. 136:2729. (Block)
  3. Suzuki K, et al. 2000. Immunity 13:691. (IHC)
  4. Shen FW, et al. 1986. Immunogenetics 24:146. (IP)
  5. Baldwin TA and Hogquist KA. 2007. J. Immunol. 179:837.
  6. Pascal V, et al. 2007. J. Immunol. 179:1751.
  7. Burman AC, et al. 2007. Blood 110:1064.
  8. Kincaid EZ, et al. 2007. J. Immunol. 179:3187.
  9. Phan TG, et al. 2007. Nature Immunol. 8:992.
  10. Nakano-Yokomizo T, et al. 2011. J. Exp Med. 208:1661. PubMed
  11. Wen T, et al. 2013. PNAS. 110:6067. PubMed
  12. Kohlmeier JE, et al. 2008. Immunity. 29:101. (FC) PubMed
Product Citations
  1. Faulhaber LD, et al. 2022. Neurophotonics. 9:031917. PubMed
RRID
AB_2629589 (BioLegend Cat. No. 109850)

Antigen Details

Structure
Protein tyrosine phosphatase (PTP) family, 180-240 kD
Distribution

All hematopoietic cells except mature erythrocytes and platelets of the CD45.2 strain of mice

Function
Phosphatase, T and B cell activation
Ligand/Receptor
Galectin-1, CD2, CD3, CD4
Biology Area
Cell Biology, Immunology, Inhibitory Molecules, Innate Immunity, Neuroscience, Neuroscience Cell Markers
Molecular Family
CD Molecules
Antigen References

1. Suzuki K, et al. 2000. Immunity 13:691.

Gene ID
19264 View all products for this Gene ID
UniProt
View information about CD45.2 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.

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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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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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