Brilliant Violet 421™ anti-mouse CD45 Antibody

Pricing & Availability
Clone
30-F11 (See other available formats)
Regulatory Status
RUO
Other Names
T200, Ly-5, LCA
Isotype
Rat IgG2b, κ
Ave. Rating
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Product Citations
publications
30-F11_BV421_051911
C57BL/6 mouse splenocytes were stained with CD45 (clone 30-F11) Brilliant Violet 421™ (filled histogram) or rat IgG2b Brilliant Violet 421™ isotype control (open histogram).
  • 30-F11_BV421_051911
    C57BL/6 mouse splenocytes were stained with CD45 (clone 30-F11) Brilliant Violet 421™ (filled histogram) or rat IgG2b Brilliant Violet 421™ isotype control (open histogram).
Compare all formats See Brilliant Violet 421™ spectral data
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103133 125 µL 146€
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103134 50 µg 194€
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Description

CD45 is a 180-240 kD glycoprotein also known as the leukocyte common antigen (LCA), T200, or Ly-5. It is a member of the protein tyrosine phosphatase (PTP) family, expressed on all hematopoietic cells except mature erythrocytes and platelets. There are different isoforms of CD45 that arise from variable splicing of exons 4, 5, and 6, which encode A, B, and C determinants, respectively. CD45 plays a key role in TCR and BCR signal transduction. These isoforms are very specific to the activation and maturation state of the cell as well as 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
Rat
Immunogen
Mouse thymus or spleen
Formulation
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 Brilliant Violet 421™ under optimal conditions.
Concentration
µg sizes: 0.2 mg/mL
µL 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 - 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 immunofluorescent staining using the µg size, the suggested use of this reagent is ≤0.25 µg per million cells in 100 µl volume. For immunofluorescent staining using the µl size, 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. It is recommended that the reagent be titrated for optimal performance for each application.

Brilliant Violet 421™ excites at 405 nm and emits at 421 nm. The standard bandpass filter 450/50 nm is recommended for detection. Brilliant Violet 421™ is a trademark of Sirigen Group Ltd.


Learn more about Brilliant Violet™.

This product is subject to proprietary rights of Sirigen Inc. and is made and sold under license from Sirigen Inc. The purchase of this product conveys to the buyer a non-transferable right to use the purchased product for research purposes only. This product may not be resold or incorporated in any manner into another product for resale. Any use for therapeutics or diagnostics is strictly prohibited. This product is covered by U.S. Patent(s), pending patent applications and foreign equivalents.
Excitation Laser
Violet Laser (405 nm)
Application Notes

Clone 30-F11 reacts with all isoforms and both CD45.1 and CD45.2 alloantigens of CD45.

Additional reported applications (for relevant formats) include: immunoprecipitation3, complement-dependent cytotoxicity1,5, immunohistochemistry (acetone-fixed frozen sections, zinc-fixed paraffin-embedded sections and formalin-fixed paraffin-embedded sections)4,6, Western blotting7, and spatial biology (IBEX)10,11. The Ultra-LEAF™ purified antibody (Endotoxin < 0.01 EU/µg, Azide-Free, 0.2 µm filtered) is recommended for functional assays (Cat. No. 103163 and 103164).

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).

Application References

(PubMed link indicates BioLegend citation)
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  7. Tsuboi S and Fukuda M. 1998. J. Biol. Chem. 273:30680. (WB) PubMed
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  10. Radtke AJ, et al. 2020. Proc Natl Acad Sci U S A. 117:33455-65. (SB) PubMed
  11. Radtke AJ, et al. 2022. Nat Protoc. 17:378-401. (SB) PubMed
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RRID
AB_10899570 (BioLegend Cat. No. 103133)
AB_2562559 (BioLegend Cat. No. 103134)

Antigen Details

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

All hematopoietic cells except mature erythrocytes and platelets

Function
Phosphatase, T and B cell activation
Ligand/Receptor
Galectin-1, CD2, CD3, CD4, TCR, CD22, Thy-1
Cell Type
B cells, Dendritic cells, Mesenchymal Stem Cells, Tregs
Biology Area
Cell Biology, Immunology, Inhibitory Molecules, Innate Immunity, Neuroscience, Neuroscience Cell Markers, Stem Cells
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
UniProt
View information about CD45 on UniProt.org

Related FAQs

What is the F/P ratio range of our BV421™ format antibody reagents?

It is lot-specific. On average it ranges between 2-4.

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 CD45 Reagents Request Custom Conjugation
Description Clone Applications
APC anti-mouse CD45 30-F11 FC
Biotin anti-mouse CD45 30-F11 FC
FITC anti-mouse CD45 30-F11 FC
PE anti-mouse CD45 30-F11 FC
PE/Cyanine5 anti-mouse CD45 30-F11 FC
Purified anti-mouse CD45 30-F11 FC,IHC-F,CyTOF®,IP,CMCD,IHC,WB
PE/Cyanine7 anti-mouse CD45 30-F11 FC
APC/Cyanine7 anti-mouse CD45 30-F11 FC
Alexa Fluor® 488 anti-mouse CD45 30-F11 FC,SB
Alexa Fluor® 647 anti-mouse CD45 30-F11 FC,ICC,IHC,3D IHC,SB
Pacific Blue™ anti-mouse CD45 30-F11 FC
Alexa Fluor® 700 anti-mouse CD45 30-F11 FC,SB
PerCP/Cyanine5.5 anti-mouse CD45 30-F11 FC
PerCP anti-mouse CD45 30-F11 FC
Alexa Fluor® 594 anti-mouse CD45 30-F11 IHC-F,FC,3D IHC
Brilliant Violet 421™ anti-mouse CD45 30-F11 FC,SB
Brilliant Violet 570™ anti-mouse CD45 30-F11 FC
Brilliant Violet 510™ anti-mouse CD45 30-F11 FC
Brilliant Violet 605™ anti-mouse CD45 30-F11 FC
Purified anti-mouse CD45 (Maxpar® Ready) 30-F11 FC,CyTOF®
PE/Dazzle™ 594 anti-mouse CD45 30-F11 FC
Brilliant Violet 711™ anti-mouse CD45 30-F11 FC
Brilliant Violet 785™ anti-mouse CD45 30-F11 FC
Brilliant Violet 650™ anti-mouse CD45 30-F11 FC
APC/Fire™ 750 anti-mouse CD45 30-F11 FC
Brilliant Violet 750™ anti-mouse CD45 30-F11 FC
TotalSeq™-A0096 anti-mouse CD45 30-F11 PG
TotalSeq™-B0096 anti-mouse CD45 30-F11 PG
Ultra-LEAF™ Purified anti-mouse CD45 30-F11 FC,CyTOF®,IP,CMCD,IHC,WB
Spark Blue™ 550 anti-mouse CD45 30-F11 FC
Spark NIR™ 685 anti-mouse CD45 30-F11 FC
TotalSeq™-C0096 anti-mouse CD45 30-F11 PG
Spark YG™ 570 anti-mouse CD45 30-F11 IHC-F
PE/Fire™ 640 anti-mouse CD45 30-F11 FC
APC/Fire™ 810 anti-mouse CD45 30-F11 FC
PE/Fire™ 700 anti-mouse CD45 30-F11 FC
Spark Violet™ 538 anti-mouse CD45 30-F11 FC
Spark YG™ 593 anti-mouse CD45 30-F11 FC
Spark Blue™ 574 anti-mouse CD45 Antibody 30-F11 FC
Spark Blue™ 515 anti-mouse CD45 30-F11 FC
Spark UV™ 387 anti-mouse CD45 30-F11 FC
PE/Fire™ 810 anti-mouse CD45 30-F11 FC
Spark Red™ 718 anti-mouse CD45 (Flexi-Fluor™) 30-F11 FC
Spark PLUS UV™ 395 anti-mouse CD45 30-F11 FC
Go To Top Version: 3    Revision Date: 04/20/2022

For Research Use Only. Not for diagnostic or therapeutic use.

 

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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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