Alexa Fluor® 488 anti-mouse CD172a (SIRPα) Antibody

Pricing & Availability
Clone
P84 (See other available formats)
Regulatory Status
RUO
Other Names
SHPS-1, BIT, P84, PTPNS1, CD172 antigen-like family member A
Isotype
Rat IgG1, κ
Ave. Rating
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Product Citations
publications
1_P84_A488_CD172a_Antibody_1_020617
C57BL/6 mouse bone marrow cells were stained with CD11b PE and CD172a (clone P84) Alexa Fluor™ 488 (top) or rat IgG1, κ Alexa Fluor™ 488 isotype control (bottom). Data shown was gated on myeloid cell population.
  • 1_P84_A488_CD172a_Antibody_1_020617
    C57BL/6 mouse bone marrow cells were stained with CD11b PE and CD172a (clone P84) Alexa Fluor™ 488 (top) or rat IgG1, κ Alexa Fluor™ 488 isotype control (bottom). Data shown was gated on myeloid cell population.
  • 2_P84_A488_CD172a_Antibody_2_020617
  • 3_38_Mouse_Thymus_DEC205_SIRPa
    Confocal image of C57BL/6 mouse thymus sample acquired using the IBEX method of highly multiplexed antibody-based imaging: DEC205 (magenta) in Cycle 2 and SIRPα (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).
  • 4_58_Mouse_Lymph_Node_gp38_SIRPa
    Mice were injected subcutaneously with sheep red blood cells in a volume of 25 µl per site on days 0 and 4 and harvested on day 11. Confocal image of C57BL/6 mouse lymph node acquired using the IBEX method of highly multiplexed antibody-based imaging: gp38 (purple) in Cycle 2 and SIRPα (cyan) in Cycle 5. 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® 488 spectral data
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144023 25 µg $136
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144024 100 µg $347
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Description

CD172a, also known as SIRPα, is a type I transmembrane protein with one V-set Ig-like and two C-set Ig-like domains in the extracellular portion, and two ITIM motifs and a proline-rich region in the cytoplasmic tail. CD172a is expressed by monocytes, macrophages, myeloid cells, and neuronal tissue. The phosphorylation of SIRPα ITIMs induces the recruitment and activation of the tyrosine phosphatases PTPN6 and PTPN11, resulting in the negative regulation of several biological processes. The ligands of CD172a are CD47, SP-A, and SP-D.

Product Details
Technical data sheet

Product Details

Verified Reactivity
Mouse
Antibody Type
Monoclonal
Host Species
Rat
Immunogen
Mouse brain membrane protein
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® 488 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. For flow cytometric staining, the suggested use of this reagent is ≤0.5 µg per million cells in 100 µl volume. It is recommended that the reagent be titrated for optimal performance for each application.

* Alexa Fluor® 488 has a maximum emission of 519 nm when it is excited at 488 nm.


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

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Excitation Laser
Blue Laser (488 nm)
Application Notes

Additional reported applications (for the relevant formats) include: blocking SIRPa interaction with CD474, in vivo blocking of dendritic cell migration3, enhancing of macrophage phagocytosis2,4, immunohistochemical staining of cerebellum frozen sections1, immunoprecipitation2,4, and spatial biology (IBEX)5,6.

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)
  1. Comu S, et al. 1997. J. Neurosci. 17:8702. (IHC)
  2. Gresham HD, et al. 2000. J. Exp. Med. 191:515. (IP)
  3. Fukunaga A, et al. 2004. J. Immunol. 172:4091. (Block)
  4. Oldenborg PA, et al. 2000. Science 288:2051. (Block, IP)
  5. Radtke AJ, et al. 2020. Proc Natl Acad Sci U S A. 117:33455-65. (SB) PubMed
  6. Radtke AJ, et al. 2022. Nat Protoc. 17:378-401. (SB) PubMed
Product Citations
  1. Clement CC, et al. 2021. Immunity. 54:721. PubMed
RRID
AB_2650814 (BioLegend Cat. No. 144023)
AB_2650815 (BioLegend Cat. No. 144024)

Antigen Details

Structure
Type I transmembrane protein
Distribution

Monocytes, macrophages, myeloid cells, neuronal tissue

Function
Negative regulation of several biological processes
Interaction
PTPN6, PTPN11
Ligand/Receptor
CD47, SP-A, SP-D
Cell Type
Dendritic cells, Macrophages, Monocytes, Neutrophils
Biology Area
Cell Adhesion, Cell Biology, Immunology, Signal Transduction
Molecular Family
Adhesion Molecules, CD Molecules, Protein Kinases/Phosphatase
Antigen References

1. Zhao XW, et al. 2011. P. Natl. Acad. Sci. USA 108:18342.
2. Verjan-Garcia N, et al. 2011. J. Immunol. 187:2268.
3. Sato-Hashimoto M, et al. 2011. J. Immunol. 187:291.
4. Raymond M, et al. 2010. Eur. J. Immunol. 40:3510.

Gene ID
19261 View all products for this Gene ID
UniProt
View information about CD172alpha; 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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*These products may be covered by one or more Limited Use Label Licenses (see the BioLegend Catalog or our website, www.biolegend.com/ordering#license). BioLegend products may not be transferred to third parties, resold, modified for resale, or used to manufacture commercial products, reverse engineer functionally similar materials, or to provide a service to third parties without written approval of BioLegend. By use of these products you accept the terms and conditions of all applicable Limited Use Label Licenses. Unless otherwise indicated, these products are for research use only and are not intended for human or animal diagnostic, therapeutic or commercial 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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