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Alexa Fluor® 700 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
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Product Citations
publications
30-F11_Alx700_020608
C57BL/6 mouse splenocytes stained with 30-F11 Alexa Fluor® 700
  • 30-F11_Alx700_020608
    C57BL/6 mouse splenocytes stained with 30-F11 Alexa Fluor® 700
  • 54_Mouse_Liver_Ecadherin_CD45
    Confocal image of C57BL/6 mouse liver sample acquired using the IBEX method of highly multiplexed antibody-based imaging: E-Cadherin (red) in Cycle 1 and CD45 (cyan) in Cycle 3. Tissues were prepared using ~1% (vol/vol) formaldehyde and a detergent. Following fixation, samples are immersed in 30% (wt/vol) sucrose for cryoprotection. Images are courtesy of Drs. Andrea J. Radtke and Ronald N. Germain of the Center for Advanced Tissue Imaging (CAT-I) in the National Institute of Allergy and Infectious Diseases (NIAID, NIH).
Compare all formats See Alexa Fluor® 700 spectral data
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103127 25 µg $94
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103128 100 µg $215
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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

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.
Preparation
The antibody was purified by affinity chromatography and conjugated with Alexa Fluor® 700 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

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. The suggested use of this reagent is ≤ 0.25 µg per 106 cells in 100 µl volume. It is highly recommended that the reagent be titrated for optimal performance for each application.

* Alexa Fluor® 700 has a maximum emission of 719 nm when it is excited at 633nm / 635nm. Prior to using Alexa Fluor® 700 conjugate for flow cytometric analysis, please verify your flow cytometer's capability of exciting and detecting the fluorochrome.


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

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Excitation Laser
Red Laser (633 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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  11. Radtke AJ, et al. 2022. Nat Protoc. 17:378-401. (SB) PubMed
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RRID
AB_493714 (BioLegend Cat. No. 103127)
AB_493714 (BioLegend Cat. No. 103128)

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

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 UV395™ anti-mouse CD45 30-F11 FC
Go To Top Version: 4    Revision Date: 04/20/2022

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.

 

8999 BioLegend Way, San Diego, CA 92121 www.biolegend.com
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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