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Spark Blue™ 574 anti-mouse CD69 (Flexi-Fluor™) Antibody

Pricing & Availability
Clone
H1.2F3 (See other available formats)
Regulatory Status
RUO
Other Names
Very Early Activation Antigen (VEA), AIM, EA1, MLR3, gp34/28
Isotype
Armenian Hamster IgG
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Product Citations
publications
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Cat # Size Price Quantity Save
285173 50 µg £230
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Description

CD69 is a 60 kD type II membrane protein composed of a 27/33 kD disulfide-linked homodimer, also known as Very Early Activation Antigen (VEA), AIM, EA1, MLR3, and gp34/28. It is expressed on a subset of thymocytes and platelets. CD69 is rapidly induced on activated T and B cells, neutrophils, and NK cells. It is a C-type lectin, closely related to the NKR-P1 and Ly-49 NK cell activation molecules. CD69 is involved in the early events of cell activation and thymocyte positive selection.

Product Details
Technical Data Sheet (pdf)

Product Details

Verified Reactivity
Mouse
Antibody Type
Monoclonal
Host Species
Armenian Hamster
Immunogen
Mouse dendritic epidermal T cell line Y245
Formulation
Phosphate-buffered solution, pH 7.2, containing 0.09% sodium azide
Preparation
The antibody was purified by affinity chromatography and conjugated with Spark Blue™ 574 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

FC
Recommended Usage

Flexi-Fluors™ are provided at a standard 0.2 mg/mL concentration. We recommend titrating this reagent to determine the optimal concentration for each application. For many flow cytometry applications, conjugated antibodies perform well at concentrations ranging from 0.03 to 1.0 µg per million cells in 100 µL. We recommend testing a range of concentrations starting from 10 µg/mL.

For example, make five 1:1 serial dilutions of the 0.2 mg/mL antibody. Add 5 µL of each dilution (including the undiluted antibody) to 100 µL of cells (at 107 cells/mL) to test six concentrations -- 1.0, 0.5, 0.25, 0.125, 0.06, and 0.03 µg per million cells in 100 µL volume. Compare staining patterns or create a titration curve using the MFI or staining index to determine the optimal concentration.

Excitation Laser
Blue Laser (488 nm)
Application Notes

The H1.2F3 antibody has been reported to augment T cell activation. Additional reported applications (for the relevant formats) include: in vitro T cell and NK cell activation1-3, immunohistochemistry4,5, and immunoprecipitation1.

This antibody has been characterized in the literature as containing a lambda (?) light chain.

Additional Product Notes

For more information about Flexi-Fluors™, visit our Flexi-Fluor™ page and review FAQs associated with this product line.

Application References

(PubMed link indicates BioLegend citation)
  1. Yokoyama WM, et al. 1988. J. Immunol. 141:369. (IP)
  2. Sobel ES, et al. 1993. J. Immunol. 150:673.
  3. Karlhofer FM, et al. 1991. J. Immunol. 146:3662.
  4. Zhou X, et al. 2005. J. Biol. Chem. 280:31240. (IHC)
  5. Podd BS, et al. 2006. J. Immunol. 176:6532. (IHC)
  6. Lawson BR, et al. 2007. J. Immunol. 178:5366.
  7. Lee JW, et al. 2006. Nature Immunol. 8:181.
  8. Epardaud M, et al. 2008. Cancer Res. 15:2972. PubMed
  9. Jordan JM, et al. 2008. 76:3717. PubMed
  10. Kenna TJ, et al. 2008. Blood 111:2091. PubMed
  11. Ishikawa C, et al. 2013. Biochim Biophys Acta. 167:99. PubMed
RRID
AB_3662513 (BioLegend Cat. No. 285173)

Antigen Details

Structure
C-type lectin, 27/33 kD
Distribution

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

Function
Lymphocyte activation
Cell Type
B cells, Granulocytes, NK cells, Platelets, T cells, Thymocytes, Tregs
Biology Area
Costimulatory Molecules, Immunology, Innate Immunity
Molecular Family
CD Molecules
Antigen References

1. Barclay AN, et al. 1997. The Leukocyte Antigen FactsBook Academic Press.
2. Testi R, et al. 1994. Immunol. Today 15:479.
3. Moretta A, et al. 1991. J. Exp. Med. 174:1393.
4. Yokoyama WM, et al. 1988. J. Immunol. 141:369.

Gene ID
NA
UniProt
View information about CD69 on UniProt.org

Related FAQs

What are Flexi-Fluors?

Flexi-Fluors are rapidly made-to-order conjugated antibodies. The technology, manufacturing processes, and specifications used to create Flexi-Fluors are the same as our regular catalog products. However, the optimal concentration and performance of each Flexi-Fluor must be determined by the customer. 

How quickly will I receive my order?

We aim to ship Flexi-Fluors within 2-3 weeks of receipt of your order. However, depending on your location, shipping times may vary. 

How are Flexi-Fluors different from regular catalog products?

Flexi-Fluors are made on demand, specifically for you. Flexi-Fluors are manufactured using the same high-quality standards, and specifications as other catalog products. For faster delivery, Flexi-Fluors are not tested by flow cytometry to determine optimal concentrations or evaluate performance. This testing needs to be performed by the customer.

How do I determine the optimal concentration for using my Flexi-Fluor? How should I titrate my antibody?

Flexi-Fluors are provided at a standard 0.2 mg/mL concentration. We recommend that you titrate your antibody to determine the optimal concentration to use for your application. For many flow cytometry applications, conjugated antibodies perform well at concentrations ranging from 0.03 to 1.0 µg per million cells in 100 µL volume. We recommend that you test a range of concentrations starting from 10 µg/mL. 

 

For example, make five 1:1 serial dilutions of your 0.2 mg/mL antibody.  Add 5 µL of each dilution (including the undiluted antibody) to 100 µL of cells (at 107 cells/ml) to test six concentrations - 1.0, 0.5, 0.25, 0.125, 0.06, and 0.03 µg per million cells in 100 µL volume. Compare staining patterns or create a titration curve using the MFI or staining index to determine the optimal concentration. 

I can’t find the antibody-dye combination that I need. When will it be available?

We continuously update our catalog, introducing scores of new products every month. Please get in touch with our Technical Service team for an update on new products or recommendations for suitable alternatives to complete your panel. Or contact Custom Solutions to inquire about our affordable custom conjugation services. 

I need help to validate the performance of my Flexi-Fluor. Who should I contact?

Please get in touch with Technical Service for assistance. 

Can I order more than 50 μg of a Flexi-Fluor?

Yes, you can order multiple vials of the same Flexi-Fluor products.  We cannot guarantee, however, that these vials will be bottled from the same lot. For bulk single-lot orders, contact our Custom Solutions team.

What is the expiration date of my Flexi-Fluor?

Expiration dates can be found on the vial label or by using our CoA lookup tool. 

Go To Top Version: 1    Revision Date: 09/06/2024

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.

 

BioLegend, the BioLegend logo, and all other trademarks are property of BioLegend, Inc. or their respective owners, and all rights are reserved.

 

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Toll-Free Phone: 1-877-Bio-Legend (246-5343) Phone: (858) 768-5800 Fax: (877) 455-9587

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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