Product Information

Contents: Alexa Fluor® 647 anti-mouse/human T-Bet
Catalog Number: 51-5825
Sizes: 25 ug
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.
Clone: eBio4B10 (4B10, 4-B10)
Isotype: Mouse IgG1

CD4+ T cells were sorted from mouse spleen, cultured in either Th1- (left) or Th2- (right) polarizing conditions and restimulated with PMA and Ionomycin. Th1 or Th2 cells were then stained intracellularly, using the eBioscience Foxp3 Staining Buffer Set (cat. 00-5523), with 1.0 μg of Alexa Fluor® 647 Mouse IgG1, K Iso Cntrl (cat. 51-4714) (open histogram) or 1.0 μg of Alexa Fluor® 647 anti-mouse/human T-Bet (colored histogram). Total cells were used for analysis.

Available Formats of This Product
Cat. No.	Format	Excite (nm)	Emit (nm)	Reported Applications
14-5825	Affinity Purified anti-mouse/human T-Bet (Tbet, Th1-specific T box transcription factor)	N/A	N/A	FC IP WB
12-5825	Phycoerythrin (PE) anti-mouse/human T-Bet (Tbet, Th1-specific T box transcription factor)	488	575	IC Flow
51-5825	Alexa Fluor® 647 anti-mouse/human T-Bet (Tbet, Th1-specific T box transcription factor)	633	668	IC Flow
45-5825	PerCP-Cy5.5 anti-mouse/human T-Bet (Tbet, Th1-specific T box transcription factor) (PE-Cy5.5 replacement)	488	690	IC Flow
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.

Custom Product Requests
Need a custom product? Download the Custom Product Request Form and submit completed form to customs@ebioscience.com.

Questions? Please consult our answers to frequently asked questions at http://www.ebioscience.com/faq.

Description

The eBio4B10 monoclonal antibody reacts with mouse and human T-bet. T-bet is a Th1-specific T-box transcription factor critical to the development of the Th1 CD4+ T cell lineage. This is known based on the observations that T-Bet deficient mice have impaired Th1 cell development, and that ectopic expression of T-Bet results in development skewed to the Th1 lineage. T-Bet expression is induced by the Th1 cytokine IFN-γ, and T-Bet also regulates the expression of IFN-γ, likely, at least in part, through the modification of DNA accessibility and histone remodeling. In addition to IFN-γ, T-Bet is also known to regulate the expression of IL-12Rβ and IL-2. Moreover, T-Bet plays a role in class-switch recombination in B-cells.

Applications Reported

For research use only, not for diagnostic or therapeutic use. This eBio4B10 (4B10, 4-B10) antibody has been reported for use in intracellular staining followed by flow cytometric analysis.

Applications Tested

This eBio4B10 (4B10, 4-B10) antibody has been tested by intracellular staining and flow cytometric analysis of Th1-polarized CD4+ T cells. This can be used at less than or equal to 2 μg per 100 μl blood (or per 1 million cells in 100 μl total staining volume). It is recommended that the antibody be carefully titrated for optimal performance in the assay of interest.

For intracellular staining with the eBio4B10 monoclonal antibody, it is recommended to use the eBioscience Foxp3 Staining Buffer Set (cat. 00-5523).

References

Szabo SJ, Kim ST, Costa GL, Zhang X, Fathman CG, Glimcher LH. A novel transcription factor, T-bet, directs Th1 lineage commitment. Cell. 2000 Mar 17;100(6):655-69. (4B10, FC, WB, PubMed)

Usui T, Preiss JC, Kanno Y, Yao ZJ, Bream JH, O'Shea JJ, Strober W. T-bet regulates Th1 responses through essential effects on GATA-3 function rather than on IFNG gene acetylation and transcription. J Exp Med. 2006 Mar 20;203(3):755-66.

Hwang ES, Hong JH, Glimcher LH. IL-2 production in developing Th1 cells is regulated by heterodimerization of RelA and T-bet and requires T-bet serine residue 508. J Exp Med. 2005 Nov 7;202(9):1289-300.

Related Products

Cat. 51-4714 Alexa Fluor® 647 Mouse IgG1, K Isotype Control

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.