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PE anti-mouse CD169 (Siglec-1) Antibody

Pricing & Availability
Clone
3D6.112 (See other available formats)
Regulatory Status
RUO
Other Names
Sialic acid binding Ig-like lectin 1 (Siglec-1), Sialoadhesin (Sn)
Isotype
Rat IgG2a, κ
Ave. Rating
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Product Citations
publications
1_3D6.112_PE_1_110811
C57BL/6 splenocytes were stained with F4/80 APC, Ly-6G PerCP, and CD169 (clone 3D6.112) PE (top) or rat IgG2a, κ PE isotype control (bottom). Data was analyzed by gating on Ly-6G-negative cell population.
  • 1_3D6.112_PE_1_110811
    C57BL/6 splenocytes were stained with F4/80 APC, Ly-6G PerCP, and CD169 (clone 3D6.112) PE (top) or rat IgG2a, κ PE isotype control (bottom). Data was analyzed by gating on Ly-6G-negative cell population.
  • 2_3D6.112_PE_2_110811
  • 3_Yu_Kato
    C57BL/6 inguinal lymph node labeled in-vivo with PE anti-mouse CD169 (red, clone 3D6.112), followed by intravital imaging of the single harmonic generation of the collagen fibers of LN capsule (blue), and CMFDA labelled polyclonal B cells (green). Image generously submitted to the 2017 Cell Life Imaging Competition by Yu Kato from La Jolla Institute for Allergy and Immunology.
  • 4_3D6dot112_PE_CD169_Antibody_IHC_1_012218
    Live intravital mouse spleen imaging. PE CD169 (red) (clone 3D6.112), Alexa Fluor® 647 F4/80 (blue) (clone BM8), and autofluorescence (green) were imaged for 30 minutes after IV injection of 10 μg per antibody. Isotype controls at the same concentrations, time post injection, and exposure conditions were used for the negative control. Cells were imaged with a Leica SP5 confocal on anesthetized mice. Image courtesy of Grzegorz Chodaczek and Zbigniew Mikulski at LIAI.
  • 5_3D6dot112_PE_CD169_Antibody_IHC_2_012218
    Fixed whole mount mouse spleen imaging sectioned after intravital imaging completed. PE CD169 (red) (clone 3D6.112), Alexa Fluor® 647 F4/80 (blue) (clone BM8), and autofluorescence (green) IV injected at 10 μg per antibody. Spleen imaged intravital, removed, sectioned, and imaged again with a Leica SP8 confocal. Image courtesy of Grzegorz Chodaczek and Zbigniew Mikulski at LIAI.
  • 6_33_Mouse_Spleen_CD169_CD45
    Confocal image of C57BL/6 mouse spleen sample acquired using the IBEX method of highly multiplexed antibody-based imaging: CD169 (green) in Cycle 2 and CD45 (blue) 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).
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142403 25 µg $153
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142404 100 µg $360
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Description

CD169, also known as Siglec-1 and Sialoadhesin (Sn), is a type I lectin containing 17 immunoglobulin (Ig) domains (one variable domain and 16 constant domains). CD169 binds to sialic acids, which can be found on PSGL-1, CD43, CD206, and CD227. By its affinity to α2, 3-linked sialic acid, it is involved in macrophage binding to different cell types such as granulocytes, monocytes, NK, B, and T cells. CD169 was initially identified as a sialic acid-dependent sheep erythrocyte receptor (SER) on resident bone marrow cells of mice. It has been identified as highly expressed on resident bone marrow macrophages which plays an important role in retention of stem cells in mesenchymal stem cell niche. It is also found on some specific subsets of tissue macrophages in spleen, lymph nodes, bone marrow, liver, colon, lungs, and cancer cells. Evidence suggest that CD169-positive macrophages serve as lymph node-resident APCs to dominate early activation of tumor antigen-specific CD8+ T cells and invariant NK cell.

Product Details
Technical data sheet

Product Details

Verified Reactivity
Mouse
Antibody Type
Monoclonal
Host Species
Rat
Immunogen
Purified Native Sialoadhesin from spleen
Formulation
Phosphate-buffered solution, pH 7.2, containing 0.09% sodium azide.
Preparation
The antibody was purified by affinity chromatography and conjugated with PE under optimal conditions.
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.

Excitation Laser
Blue Laser (488 nm)
Green Laser (532 nm)/Yellow-Green Laser (561 nm)
Application Notes

Additional reported applications (for the relevant formats) include: immunohistochemical staining in frozen tissue sections1, and spatial biology (IBEX)4,5.

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. Barral P, et al. 2010. Nat. Immunol. 11:303. (IHC-F)
  2. Chtanova T, et al. 2008. Immunity 29:487.
  3. Klass M, et al. 2012. J. Immunol. 189:2414. PubMed
  4. Radtke AJ, et al. 2020. Proc Natl Acad Sci U S A. 117:33455-65. (SB) PubMed
  5. Radtke AJ, et al. 2022. Nat Protoc. 17:378-401. (SB) PubMed
Product Citations
  1. Voss LF, et al. 2023. Front Immunol. 13:1021370. PubMed
  2. Fernández-García V, et al. 2022. Biomed Pharmacother. 148:112769. PubMed
  3. Anania JC, et al. 2021. Front Immunol. 12:655753. PubMed
  4. Farsakoglu Y et al. 2019. Cell reports. 26(9):2307-2315 . PubMed
  5. Juul-Madsen K, et al. 2021. Proc Natl Acad Sci U S A. 118:. PubMed
  6. Uchil PD et al. 2018. Cell host & microbe. 25(1):87-100 . PubMed
  7. Kato Y, et al. 2020. Immunity. 53(3):548-563. PubMed
  8. Leylek R, et al. 2019. Cell Rep. 29:3736. PubMed
  9. Tacconi C, et al. 2021. Cell Reports. 35(2):108993. PubMed
  10. Tomay F, et al. 2019. J Transl Med. 17:237. PubMed
  11. Beura LK, et al. 2018. Immunity. 48:327. PubMed
  12. Leary N, et al. 2022. J Extracell Vesicles. 11:e12197. PubMed
  13. Feuerstein R, et al. 2015. J Immunol. 194:2735. PubMed
  14. Song X, et al. 2022. Transl Oncol. 15:101306. PubMed
RRID
AB_10915470 (BioLegend Cat. No. 142403)
AB_10915470 (BioLegend Cat. No. 142404)

Antigen Details

Structure
Type I single membrane-spanning lectin containing 17 immunoglobulin (Ig) domains, belongs to the immunoglobulin superfamily.
Distribution

Macrophages in spleen, lymph nodes, bone marrow, liver, colon and lungs.

Function
Adhesion.
Ligand/Receptor
PSGL-1, CD43, CD206 and CD227.
Cell Type
Macrophages
Biology Area
Cell Biology, Immunology
Molecular Family
Adhesion Molecules, CD Molecules, Protein Kinases/Phosphatase, Siglec Molecules
Antigen References

1. Chow A, et al. 2011. J. Exp. Med. 208:261.
2. Asano K, et al. 2011. Immunity 34:85.
3. Xiong YS, et al. 2009. Clin. Biochem. 42:1057.
4. Varki A, et al. 2009. Glycoconj. J. 26:231.
5. Rempel H, et al. 2008. PLoS One 3:e1967.
6. Crocker PR, et al. 2001. Trends Immunol. 22:337.
7. Hartnell A, et al. 2001. Blood 97:288.
8. Crocker PR, et al. 1985. J. Exp. Med. 162:993.

Gene ID
20612 View all products for this Gene ID
UniProt
View information about CD169 on UniProt.org

Related FAQs

What type of PE do you use in your conjugates?
We use R-PE in our conjugates.
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.

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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