Product Information

Contents: Allophycocyanin-Alexa Fluor® 750 (APC-Alexa Fluor® 750) anti-human CD154 (CD40 Ligand, CD40L)
Catalog Number: 27-1548
Formulation: Phosphate buffered saline, pH 7.2, less than or equal to 0.09% NaN₃, BSA and/or gelatin for protein stabilization
Storage Conditions: Store at 4°C.
DO NOT FREEZE.
LIGHT-SENSITIVE MATERIAL.
This tandem dye is sensitive to photo-induced oxidation. Protect this vial from light during storage, handling & experimental procedures. This product is guaranteed for 6 months upon receipt when stored properly.
Clone: 24-31
Isotype: Mouse IgG1, κ
HLDA No.: N/A

Staining of unstimulated PBMCs (open histogram) and 6 hour PMA/ionomycin stimulated PBMCs (colored histogram) with APC-Alexa Fluor® 750 anti-human CD154 (24-31). Total viable cells were used for analysis.

Available Formats of This Product
Cat. No.	Format	Excite (nm)	Emit (nm)	Reported Applications
14-1548	Affinity Purified anti-human CD154 (CD40 Ligand, CD40L)	N/A	N/A	FA FC
16-1548	Functional Grade* Purified anti-human CD154 (CD40 Ligand, CD40L)	N/A	N/A	FA FC
11-1548	FITC anti-human CD154 (CD40 Ligand, CD40L)	488	518	FC
12-1548	PE anti-human CD154 (CD40 Ligand, CD40L)	488	575	FC
17-1548	Allophycocyanin (APC) anti-human CD154 (CD40 Ligand, CD40L)	633	660	FC
47-1548	Allophycocyanin-eFluor^TM 780 anti-human CD154 (CD40 Ligand, CD40L)	633	780	FC
*Functional Grade™ (FG™) Purified: Azide-free, sterile-filtered, and endotoxin < 0.001 ng/µg (unless otherwise noted). *Functional Grade™ (FG™) Biotin: Azide-free, sterile-filtered, and endotoxin < 0.05 ng/µg. Purified: Contains azide, not sterile-filtered, and not endotoxin tested.
Flow Cytometry Product Notes: Test Sizes: To accommodate multicolor flow cytometry, eBioscience is in the process of reducing test size volumes from 20 µl to 5 µl. Please check your antibody vial for the recommended test size. Fluorochrome Replacements: eBioscience is in the process of replacing all Pacific Blue® and APC-Alexa Fluor® 750 conjugated products with eFluor™ 450 and APC-eFluor™ 780 conjugated products, respectively.

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Description

The 24-31 monoclonal antibody reacts with human CD154, a 39 kDa transmembrane glycoprotein also known as gp39 and CD40 ligand (CD40L). CD154 is a member of the TNF superfamily and is expressed transiently by activated T cells. Through its binding to CD40 on antigen presenting cells including B cells, monocytes/macrophages and dendritic cells, CD154 serves a crucial function in T-APC cognate interaction. CD154 interaction with CD40 transduces signals for T-dependent B-cell activation and induces B cell cycle entry. 24-31 cross-reacts with the Cynomogus monkey CD154.

Applications Reported

For research use only, not for diagnostic or therapeutic use. This 24-31 antibody has been reported for use in flow cytometric analysis.

The photo-emission properties and compensation required for Alexa Fluor® 750 are similar to APC-Cy7; however Alexa Fluor® 750 has several advantages when compared to APC-Cy7. The Alexa Fluor® 750 has peak emission of 779 nm, and usually has a slightly higher mean fluorescent intensity when compared to the same clone conjugated with APC-Cy7, which has peak emission of 767 nm. Furthermore, Alexa Fluor® 750 is considered to be a more photostable fluorochrome as compared with APC-Cy7. It is advised not to use fixatives with Alexa Fluor® 750 as this may lead to degradation of the fluorochrome and loss of staining.

Applications Tested

This 24-31 antibody has been pre-titrated and tested by flow cytometric analysis of stimulated normal human peripheral blood cells. This can be used at 20 μl (0.5 μg) per million cells in a 100 μl total staining volume.

References

Brams, P., A. Black, et al. 2001. A humanized anti-human CD154 monoclonal antibody blocks CD154-CD40 mediated human B cell activation. Int Immunopharmacol 1(2): 277-94.

Related Products

Cat. 27-4714 Allophycocyanin-Alexa Fluor® 750 (APC-Alexa Fluor® 750) Mouse IgG1, K Isotype Control (APC-Cy7 Equivalent)

Alexa Fluor® and Pacific Blue® are registered trademarks of and licensed under patents assigned to Molecular Probes, Inc. for research use only. This product is subject to an agreement between Molecular Probes, Inc. and eBioscience, and the manufacture, use, sale or import of this product may be subject to one or more U.S. patents, pending applications and corresponding foreign equivalents, owned by Molecular Probes, Inc. (a wholly owned subsidiary of Invitrogen Corp). The purchase of this product conveys to the buyer the non-transferable right to use the purchased amount of the product for life science research or as an ASR. The buyer cannot use this product for manufacturing or for any other screening (specifically including use in combination with microarrays or High Content Screening) or testing purpose, other than as an ASR. For information on purchasing a license to this product for purposes other than life science research or use as an ASR, contact Molecular Probes, Inc.