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Brilliant Violet 421™ anti-mouse CD68 Antibody

Pricing & Availability
Clone
FA-11 (See other available formats)
Regulatory Status
RUO
Other Names
Macrosialin
Isotype
Rat IgG2a
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Product Citations
publications
1_FA-11_BV421_CD68_Antibody_FC_123113
Thioglycolate-elicited BALB/c macrophages were fixed, permeabilized, and then stained with CD68 (clone FA-11) Brilliant Violet 421™ (filled histogram) or rat IgG2a, κ Brilliant Violet 421™ isotype control (open histogram).
  • 1_FA-11_BV421_CD68_Antibody_FC_123113
    Thioglycolate-elicited BALB/c macrophages were fixed, permeabilized, and then stained with CD68 (clone FA-11) Brilliant Violet 421™ (filled histogram) or rat IgG2a, κ Brilliant Violet 421™ isotype control (open histogram).
  • 36_Mouse_Spleen_CD11c_CD68
    Confocal image of C57BL/6 mouse spleen sample acquired using the IBEX method of highly multiplexed antibody-based imaging: CD11c (cyan) in Cycle 2 and CD68 (red) 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).
  • 39_Mouse_Thymus_CD68_MHCII
    Confocal image of C57BL/6 mouse thymus sample acquired using the IBEX method of highly multiplexed antibody-based imaging: CD68 (cyan) in Cycle 3 and MHCII (red) 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).
  • 46_Mouse_Lung_Btub3_CD68_CD44
    Confocal image of C57BL/6 mouse lung sample acquired using the IBEX method of highly multiplexed antibody-based imaging: CD11c (magenta) in Cycle 1, CD206 (blue) in Cycle 1, and CD31 (green) in Cycle 4. 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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137017 50 µg $275
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Description

Mouse CD68, also known as macrosialin, is an 85-115 kD member of the lysosomal-associated membrane protein (LAMP) family. It is a heavily glycosylated and predominantly intracellular protein, mainly in late endosomes. Macrosialin is the murine homolog to the human macrophage glycoprotein CD68. It is expressed on tissue macrophages, Langerhans cells and at low levels on dendritic cells. Lamp proteins may have functions relating to cell-cell interaction or cell-ligand interaction. The biological function of CD68 is not completely understood.

Product Details
Technical data sheet

Product Details

Verified Reactivity
Mouse
Antibody Type
Monoclonal
Host Species
Rat
Immunogen
Purified Con A receptor glycoproteins from the P815 cell line
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
0.2 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

ICFC - Quality tested
FC - Verified

SB - Reported in the literature, not verified in house
Recommended Usage

Each lot of this antibody is quality control tested by intracellular 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.

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 (for relevant formats) applications include: immunoprecipitation1, 2, Western Blot1, 2, immunohistochemical staining of frozen sections2 and paraformaldehyde-fixed paraffin-embedded sections3, and spatial biology (IBEX)9,10.

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. Silva RP, et al. 1999. Biochem. J. 338:687. (IP, WB)
  2. Rabinowitz SS, et al. 1991. J. Exp. Med. 174:827. (IP, WB, IHC)
  3. Wu J, et al. 2008. P. Natl. Acad. Sci. USA 105:16934. (IHC)
  4. Kayama H, et al. 2012. PNAS. 109:5010. PubMed
  5. Park S, et al. 2013. Biomaterials. 34:598. PubMed
  6. Guiducci C, et al. 2013. J Exp Med. 210:2903. PubMed
  7. McKinstry SU, et al. 2014. J Neurosci. 34:9455. PubMed
  8. Li X, et al. 2015. J Am Heart Assoc. 6:4. PubMed
  9. Radtke AJ, et al. 2020. Proc Natl Acad Sci U S A. 117:33455-65. (SB) PubMed
  10. Radtke AJ, et al. 2022. Nat Protoc. 17:378-401. (SB) PubMed
Product Citations
  1. Chen J, et al. 2022. Adv Sci (Weinh). 9:e2200668. PubMed
  2. Gomez-Salinero JM, et al. 2022. Cell Stem Cell. 29:593. PubMed
  3. Bonifacio JPPL, et al. 2022. J Virol. 96:e0087122. PubMed
  4. Cai B, et al. 2021. Mol Cancer. 20:165. PubMed
  5. Socodato R, et al. 2020. Sci Signal. 13: . PubMed
  6. Gangoso E, et al. 2021. Cell. 184:2454. PubMed
RRID
AB_2562949 (BioLegend Cat. No. 137017)

Antigen Details

Structure
A member of the lysosomal-associated membrane protein (lamp) family.
Distribution

Expressed on tissue macrophages, Langerhans cells, and at low levels on dendritic cells.

Function
Involved in cell-cell interaction or cell-ligand interaction, still not completely understood.
Cell Type
Antigen-presenting cells, Dendritic cells, Langerhans cells, Leukocytes, Macrophages
Biology Area
Cell Biology, Immunology, Innate Immunity, Neuroscience, Neuroscience Cell Markers
Molecular Family
Adhesion Molecules, CD Molecules, Innate Immune Signaling
Antigen References

1. Ramprasad MP, et al. 1996. Proc. Natl. Acad. Sci. USA 93:14833.
2. Smith MJ, et al. 1987. J. Cell. Sci. 87:113.

Gene ID
12514 View all products for this Gene ID
UniProt
View information about CD68 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.

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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