Alexa Fluor® 647 anti-human CD69 Antibody

Pricing & Availability
Clone
FN50 (See other available formats)
Regulatory Status
RUO
Workshop
IV A91
Other Names
Very Early Activation Antigen (VEA), Activation inducer molecule (AIM)
Isotype
Mouse IgG1, κ
Ave. Rating
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Product Citations
publications
1_FN50_Alx647_070207
PMA+ionomycin activated human peripheral blood lymphocytes stained with FN50 Alexa Fluor® 647
  • 1_FN50_Alx647_070207
    PMA+ionomycin activated human peripheral blood lymphocytes stained with FN50 Alexa Fluor® 647
  • 2_4_Human_LN_CD69_HLA-DR
    Confocal image of human lymph node sample acquired using the IBEX method of highly multiplexed antibody-based imaging: HLA-DR (cyan) in Cycle 3, CD69 (red) 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).
  • 3_18_Human_Liver_CD163_CD49a
    Confocal image of human liver sample acquired using the IBEX method of highly multiplexed antibody-based imaging: CD163 (yellow) in Cycle 2 and CD49a (blue) 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).
Compare all formats See Alexa Fluor® 647 spectral data
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310918 100 tests £182
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Description

CD69 is a 27-33 kD type II transmembrane protein also known as activation inducer molecule (AIM), very early activation antigen (VEA), and MLR3. It is a member of the C-type lectin family, expressed as a disulfide-linked homodimer. Other members of this receptor family include NKG2, NKR-P1 CD94, and Ly49. CD69 is transiently expressed on activated leukocytes including T cells, thymocytes, B cells, NK cells, neutrophils, and eosinophils. CD69 is constitutively expressed by a subset of medullary mature thymocytes, platelets, mantle B cells, and certain CD4+ T cells in germinal centers of normal lymph nodes. CD69 is involved in early events of lymphocyte, monocyte, and platelet activation, and has a functional role in redirected lysis mediated by activated NK cells.

Product Details
Technical Data Sheet (pdf)

Product Details

Verified Reactivity
Human
Reported Reactivity
African Green, Baboon, Chimpanzee, Cynomolgus, Pigtailed Macaque, Rhesus
Antibody Type
Monoclonal
Host Species
Mouse
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 Alexa Fluor® 647 under optimal conditions.
Concentration
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
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 5 µl per million cells in 100 µl staining volume or 5 µl per 100 µl of whole blood.

* Alexa Fluor® 647 has a maximum emission of 668 nm when it is excited at 633nm / 635nm.


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

View full statement regarding label licenses
Excitation Laser
Red Laser (633 nm)
Application Notes

Additional reported applications (for the relevant formats) include: immunohistochemical staining of acetone-fixed frozen tissue sections2, immunofluorescence microscopy3, and spatial biology (IBEX)8,9.

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. Knapp WB, et al. 1989. Leucocyte Typing IV. Oxford University Press. New York.
  2. Sakkas LI, et al. 1998. Clin. and Diag. Lab. Immunol. 5:430. (IHC)
  3. Kim JR, et al. 2005. BMC Immunol. 6:3. (IF)
  4. Verjans GM, et al. 2007. P. Natl. Acad. Sci. USA 104:3496.
  5. Lu H, et al. 2009. Toxicol Sci. 112:363. (FC) PubMed
  6. Thakral D, et al. 2008. J. Immunol. 180:7431. (FC) PubMed
  7. Yoshino N, et al. 2000. Exp. Anim. (Tokyo) 49:97. (FC)
  8. Radtke AJ, et al. 2020. Proc Natl Acad Sci USA. 117:33455-33465. (SB) PubMed
  9. Radtke AJ, et al. 2022. Nat Protoc. 17:378-401. (SB) PubMed
Product Citations
  1. Willcox CR, et al. 2020. Immunity. 51(5):813-825.e4.. PubMed
  2. Greenberg J, et al. 2022. JCI Insight. Online ahead of print. PubMed
  3. Harris MJ, et al. 2021. Mol Syst Biol. 17:e10091. PubMed
  4. Leite NC, et al. 2020. Cell Reports. 32(2):107894.. PubMed
  5. Rasmussen TA, et al. 2022. Cell Rep Med. 3:100766. PubMed
  6. Li Z, et al. 2018. J Immunol. 200:1471. PubMed
  7. Pettmann J, et al. 2021. eLife. 10:00. PubMed
RRID
AB_528871 (BioLegend Cat. No. 310918)

Antigen Details

Structure
C-type lectin, type II glycoprotein, 28/32 kD
Distribution

Activated T cells, B cells, NK cells, granulocytes, thymocytes, platelets, Langerhans cells

Function
Lymphocyte, monocyte, and platelet activation, NK cell killing
Cell Type
B cells, Granulocytes, Langerhans cells, NK cells, Platelets, T cells, Thymocytes, Tregs
Biology Area
Costimulatory Molecules, Immunology
Molecular Family
CD Molecules
Antigen References

1. Schlossman S, et al. Eds. 1995. Leucocyte Typing V. Oxford University Press. New York.
2. Testi R, et al. 1994. Immunol. Today 15:479.

Gene ID
969 View all products for this Gene ID
UniProt
View information about CD69 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 CD69 Reagents Request Custom Conjugation
Description Clone Applications
Purified anti-human CD69 FN50 FC,CyTOF®,IHC-F
FITC anti-human CD69 FN50 FC
PE anti-human CD69 FN50 FC
PE/Cyanine5 anti-human CD69 FN50 FC
APC anti-human CD69 FN50 FC
APC/Cyanine7 anti-human CD69 FN50 FC
PE/Cyanine7 anti-human CD69 FN50 FC
Alexa Fluor® 488 anti-human CD69 FN50 FC
Alexa Fluor® 647 anti-human CD69 FN50 FC,SB
Pacific Blue™ anti-human CD69 FN50 FC
Alexa Fluor® 700 anti-human CD69 FN50 FC
Biotin anti-human CD69 FN50 FC
PerCP/Cyanine5.5 anti-human CD69 FN50 FC
PerCP anti-human CD69 FN50 FC
Brilliant Violet 421™ anti-human CD69 FN50 FC
Brilliant Violet 785™ anti-human CD69 FN50 FC
Brilliant Violet 650™ anti-human CD69 FN50 FC
Brilliant Violet 510™ anti-human CD69 FN50 FC
Brilliant Violet 605™ anti-human CD69 FN50 FC
Purified anti-human CD69 (Maxpar® Ready) FN50 FC,CyTOF®
PE/Dazzle™ 594 anti-human CD69 FN50 FC
Brilliant Violet 711™ anti-human CD69 FN50 FC
APC/Fire™ 750 anti-human CD69 FN50 FC
TotalSeq™-A0146 anti-human CD69 FN50 PG
TotalSeq™-B0146 anti-human CD69 FN50 PG
TotalSeq™-C0146 anti-human CD69 FN50 PG
Brilliant Violet 750™ anti-human CD69 FN50 FC
KIRAVIA Blue 520™ anti-human CD69 FN50 FC
Spark NIR™ 685 anti-human CD69 Antibody FN50 FC
PE/Fire™ 640 anti-human CD69 FN50 FC
Spark YG™ 581 anti-human CD69 FN50 FC
TotalSeq™-D0146 anti-human CD69 FN50 PG
APC anti-human CD69 FN50 FC
Spark Blue™ 550 anti-human CD69 FN50 FC
PE anti-human CD69 FN50 FC
Spark Red™ 718 anti-human CD69 FN50 FC
GMP PE anti-human CD69 FN50 FC
PE/Fire™ 810 anti-human CD69 FN50 FC
Spark PLUS UV™ 395 anti-human CD69 FN50 FC
Go To Top Version: 3    Revision Date: 04/19/2022

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.

 

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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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