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Brilliant Violet 421™ anti-mouse F4/80 Antibody

Pricing & Availability
Clone
BM8 (See other available formats)
Regulatory Status
RUO
Other Names
EMR1, Ly71
Isotype
Rat IgG2a, κ
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Product Citations
publications
1_BM8_BV421_F4slash80_Antibody_1_FC_060616
Thioglycolate-elicited Balb/c mouse peritoneal macrophages were stained with F4/80 (clone BM8) Brilliant Violet 421™ (filled histogram) or rat IgG2a, κ Brilliant Violet 421™ isotype control (open histogram).
  • 1_BM8_BV421_F4slash80_Antibody_1_FC_060616
    Thioglycolate-elicited Balb/c mouse peritoneal macrophages were stained with F4/80 (clone BM8) Brilliant Violet 421™ (filled histogram) or rat IgG2a, κ Brilliant Violet 421™ isotype control (open histogram).
  • 2_BM8_BV421_F4slash80_Antibody_2_IHCF_060616
    C57BL/6 mouse frozen spleen section was fixed with 4% paraformaldehyde (PFA) for ten minutes at room temperature and blocked with 5% FBS plus 5% rat/mouse serum for 30 minutes at room temperature. Then the section was stained with 2 µg/ml of anti-mouse F4/80 (clone BM8) Brilliant Violet 421™ (Blue) and anti-mouse CD8a (clone 53-6.7) Alexa Fluor® 647 (red) overnight at 4°C. The image was captured with a 10X objective.
  • 34_Mouse_Spleen_F480_MHCII
    Confocal image of C57BL/6 mouse spleen sample acquired using the IBEX method of highly multiplexed antibody-based imaging: MHCII (blue) in Cycle 2 and F4/80 (magenta) in Cycle 2. 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).
  • 59_Mouse_Lymph_Node_F480_CD68_NK1.1
    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: F4/80 (cyan) in Cycle 3, CD68 (blue) in Cycle 6, and NK1.1 (magenta) in Cycle 9. 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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123131 125 µL 164€
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123137 50 µg 203€
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123132 500 µL 317€
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Description

F4/80 is a 160 kD glycoprotein. It is characterized as a member of the epidermal growth factor (EGF)-transmembrane 7 (TM7) family. F4/80, also known as EMR1 or Ly71, has been widely used as a murine macrophage marker, which is expressed on the majority of tissue macrophages including peritoneal macrophages, macrophages in lung, gut, thymus and red pulp of spleen (but not on the macrophages located in T cell areas of the spleen, lymph node and Peyer's patch), Kuffer cells, Langerhans cells, and bone marrow stromal cells. F4/80 has also been shown on a subset of dendritic cells. The biological ligand of F4/80 has not been identified, but it has been reported that F4/80 is required for induction of CD8+ T cells-mediated peripheral tolerance.

Product Details
Technical data sheet

Product Details

Verified Reactivity
Mouse
Antibody Type
Monoclonal
Host Species
Rat
Immunogen
Murine macrophages
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
IHC-F - Verified

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 µl sizes, 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. For immunohistochemical staining on frozen tissue sections, the suggested use of this reagent is 2.0 µg/ml. 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

Additional reported applications (for the relevant formats) include: immunohistochemical staining of acetone-fixed frozen sections1,2 and formalin-fixed paraffin-embedded sections6,7, Western blotting, and spatial biology (IBEX)12,13.

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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  13. Radtke AJ, et al. 2022. Nat Protoc. 17:378-401. (SB) PubMed
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RRID
AB_10901171 (BioLegend Cat. No. 123131)
AB_2563102 (BioLegend Cat. No. 123137)
AB_11203717 (BioLegend Cat. No. 123132)

Antigen Details

Structure
EGF-TM7 family member, 160 kD glycoprotein
Distribution

Majority of tissue macrophages including peritoneal macrophages, macrophages in lung, gut, thymus and red pulp of spleen, Kuffer cells, Langerhans cells, bone marrow stromal cells, and a subset of dendritic cells

Function
Induction of immunological tolerance
Cell Type
Dendritic cells, Langerhans cells, Macrophages, Tregs
Biology Area
Cell Biology, Immunology, Innate Immunity, Neuroinflammation, Neuroscience
Antigen References

1. Austy JM and Gordon S. 1981. Eur. J. Immunol. 11:805.
2. Hume DA, et al. 1983. J. Exp. Med. 158:1522.
3. Ruedl C, et al. 1996. Eur. J. Immunol. 26:1801.
4. McKnight AJ, et al. 1996. J. Biol. Chem. 271:486.
5. Lin HH, et al. 2005. J. Exp. Med. 201:1615.

Gene ID
13733 View all products for this Gene ID
UniProt
View information about F4/80 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.

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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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