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Spark Blue™ 574 anti-mouse CD45.1 (Flexi-Fluor™) Antibody

Pricing & Availability
Clone
A20 (See other available formats)
Regulatory Status
RUO
Other Names
T200, Ly-5.1, LCA
Isotype
Mouse (A.SW) IgG2a, κ
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Product Citations
publications
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Cat # Size Price Quantity Save
285182 50 µg $336
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Description

CD45.1 is an alloantigen of CD45, expressed by Ly5.1 bearing mouse strains (e.g., RIII, SJL/J, STS/A, DA). 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 mice 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 types. The primary ligands for CD45 are galectin-1, CD2, CD3, CD4, TCR, CD22, and Thy-1.

Product Details
Technical data sheet

Product Details

Verified Reactivity
Mouse
Antibody Type
Monoclonal
Host Species
Mouse
Immunogen
SJL 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 Spark Blue™ 574 under optimal conditions.
Concentration
0.2 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

FC
Recommended Usage

Flexi-Fluors™ are provided at a standard 0.2 mg/mL concentration. We recommend titrating this reagent to determine the optimal concentration for each application. For many flow cytometry applications, conjugated antibodies perform well at concentrations ranging from 0.03 to 1.0 µg per million cells in 100 µL. We recommend testing a range of concentrations starting from 10 µg/mL.

For example, make five 1:1 serial dilutions of the 0.2 mg/mL antibody. Add 5 µL of each dilution (including the undiluted antibody) to 100 µL of cells (at 107 cells/mL) to test six concentrations -- 1.0, 0.5, 0.25, 0.125, 0.06, and 0.03 µg per million cells in 100 µL volume. Compare staining patterns or create a titration curve using the MFI or staining index to determine the optimal concentration.

Excitation Laser
Blue Laser (488 nm)
Application Notes

The A20 antibody does not react with leukocytes or mouse cells expressing the CD45.2 alloantigen. Additional reported applications (for relevant formats of this clone) include: immunoprecipitation3, in vitro blocking of B cell responses1,2, immunohistochemical staining of frozen sections: OCT embedded7 and acetone-fixed4-6 (direct immunofluorescence detection with fluorochrome conjugated A20 was used in (5) and (6)).

Additional Product Notes

For more information about Flexi-Fluors™, visit our Flexi-Fluor™ page and review FAQs associated with this product line.

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. Shen FW, et al. 1986. Immunogenetics 24:146. (IP)
  4. Suzuki K, et al. 2000. Immunity 13:691. (IHC-F)
  5. Werner N, et al. 2002. Arterioscler. Thromb. Vasc. Biol. 22:1567. (IHC-F)
  6. Lessner SM, et al. 2002. Am. J. Pathol. 160:2145. (FC, IHC-F)
  7. Chen CC, et al. 2005. P. Natl. Acad. Sci. USA 102:11408 (IHC-F)
  8. Pascal V, et al. 2007. J. Immunol. 179:1751. (FC)
  9. Mende I, et al. 2006. Blood 107:1383. (IHC-F, FC)
  10. Phan TG, et al. 2007. Nature Immunol. 8:992. (FC)
  11. Wither DR, et al. 2009. J. Immunol. 183:5079. PubMed
  12. Pascal V, et al.2007. J. Immunol. 179:1751. PubMed
  13. Lee SW, et al. 2009. J. Immunol. 182:6753. PubMed
  14. Takada K, et al. 2009. J. Exp Med. 206:2253. PubMed
  15. Beamer CA, et al. 2007. Am. J. Respir. Cell. Mol. Biol. 37:729. (FC) PubMed
  16. Li LX, et al. 2010. J. Immunol. 184:1728. PubMed
  17. Hosoi A, et al. 2008. Cancer Res. 68:3941. (FC) PubMed
  18. Kenna TJ, et al. 2008. Blood 111:2091. PubMed
  19. Kohlmeier JE, et al. 2008. Immunity. 29:101. (FC) PubMed
RRID
AB_3662522 (BioLegend Cat. No. 285182)

Antigen Details

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

All hematopoietic cells except mature erythrocytes and platelets of the CD45.1 strain of mice

Function
Phosphatase, T and B cell activation
Ligand/Receptor
Galectin-1, CD2, CD3, CD4
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
NA
UniProt
View information about CD45.1 on UniProt.org

Related FAQs

What are Flexi-Fluors?

Flexi-Fluors are rapidly made-to-order conjugated antibodies. The technology, manufacturing processes, and specifications used to create Flexi-Fluors are the same as our regular catalog products. However, the optimal concentration and performance of each Flexi-Fluor must be determined by the customer. 

How quickly will I receive my order?

We aim to ship Flexi-Fluors within 2-3 weeks of receipt of your order. However, depending on your location, shipping times may vary. 

How are Flexi-Fluors different from regular catalog products?

Flexi-Fluors are made on demand, specifically for you. Flexi-Fluors are manufactured using the same high-quality standards, and specifications as other catalog products. For faster delivery, Flexi-Fluors are not tested by flow cytometry to determine optimal concentrations or evaluate performance. This testing needs to be performed by the customer.

How do I determine the optimal concentration for using my Flexi-Fluor? How should I titrate my antibody?

Flexi-Fluors are provided at a standard 0.2 mg/mL concentration. We recommend that you titrate your antibody to determine the optimal concentration to use for your application. For many flow cytometry applications, conjugated antibodies perform well at concentrations ranging from 0.03 to 1.0 µg per million cells in 100 µL volume. We recommend that you test a range of concentrations starting from 10 µg/mL. 

 

For example, make five 1:1 serial dilutions of your 0.2 mg/mL antibody.  Add 5 µL of each dilution (including the undiluted antibody) to 100 µL of cells (at 107 cells/ml) to test six concentrations - 1.0, 0.5, 0.25, 0.125, 0.06, and 0.03 µg per million cells in 100 µL volume. Compare staining patterns or create a titration curve using the MFI or staining index to determine the optimal concentration. 

I can’t find the antibody-dye combination that I need. When will it be available?

We continuously update our catalog, introducing scores of new products every month. Please get in touch with our Technical Service team for an update on new products or recommendations for suitable alternatives to complete your panel. Or contact Custom Solutions to inquire about our affordable custom conjugation services. 

I need help to validate the performance of my Flexi-Fluor. Who should I contact?

Please get in touch with Technical Service for assistance. 

Can I order more than 50 μg of a Flexi-Fluor?

Yes, you can order multiple vials of the same Flexi-Fluor products.  We cannot guarantee, however, that these vials will be bottled from the same lot. For bulk single-lot orders, contact our Custom Solutions team.

What is the expiration date of my Flexi-Fluor?

Expiration dates can be found on the vial label or by using our CoA lookup tool. 

Go To Top Version: 1    Revision Date: 09/06/2024

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.

 

BioLegend, the BioLegend logo, and all other trademarks are property of BioLegend, Inc. or their respective owners, and all rights are reserved.

 

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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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