Invitrogen™ T-bet Monoclonal Antibody (eBio4B10 (4B10)), eFluor™ 660, eBioscience™, Invitrogen™

Catalog No.	Quantity
50-112-9442	100 μg
50-165-97	25 μg

Catalog No. 50-112-9442 Supplier Invitrogen™ Supplier No. 50582582

Description

Mouse Monoclonal Antibody

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 gamma, and T-Bet also regulates the expression of IFN gamma, likely, at least in part, through the modification of DNA accessibility and histone remodeling. In addition to IFN gamma, T-Bet is also known to regulate the expression of IL-12R beta and IL-2. Moreover, T-Bet plays a role in class-switch recombination in B-cells. Applications Reported: This eBio4B10 antibody has been reported for use in intracellular staining followed by flow cytometric analysis. Applications Tested: This eBio4B10 antibody has been tested by intracellular staining and flow cytometric analysis of normal human peripheral blood cells. This can be used at less than or equal to 0.5 μg per test. A test is defined as the amount (μg) of antibody that will stain a cell sample in a final volume of 100 μL. Cell number should be determined empirically but can range from 10^5 to 10^8 cells/test. It is recommended that the antibody be carefully titrated for optimal performance in the assay of interest.

T-bet, also known as TBX21, is a T-cell specific transcription factor containing a single T-box DNA-binding domain. It controls the expression of the TH1 cytokine, interferon-gamma. Both Th1 and Th17 cells are crucial in immune regulation and autoimmune disease development. T-bet initiates Th1 lineage development from naive Th precursor cells and directs T cell differentiation to Th1 versus Th17. T-bet is cooperating with Stat4 (signal tranducer and activator of transcription 4) in programming chromatin architecture for Th1 gene expression. Genetic variations in T-bet are associated with susceptibility to aspirin-induced asthma and nasal polyps.

Specifications

• Antigen: T-bet
• Applications: Flow Cytometry
• Classification: Monoclonal
• Clone: eBio4B10 (4B10)
• Concentration: 0.2 mg/mL
• Conjugate: eFluor 660
• Formulation: PBS with 0.09% sodium azide; pH 7.2
• Gene: Tbx21
• Gene Accession No.: Q9JKD8, Q9UL17
• Gene Alias: HGNC:11599; LOW QUALITY PROTEIN: T-box transcription factor TBX21; Tbet; T-bet; TBLYM; T-box 21; T-box expressed in T cells; T-box protein 21; T-box transcription factor 21; T-box transcription factor TBX21; TBT1; TBX21; T-cell-specific T-box transcription factor T-bet; T-PET; transcription factor TBLYM; transcription factor T-box 21
• Gene Symbols: Tbx21
• Host Species: Mouse
• Purification Method: Affinity chromatography
• Quantity: 100 μg
• Regulatory Status: RUO
• Primary or Secondary: Primary
• Gene ID (Entrez): 30009, 57765, 694044
• Target Species: Human, Mouse, Rhesus Macaque
• Content And Storage: 4°C, store in dark, DO NOT FREEZE!
• Product Type: Antibody
• Form: Liquid
• Isotype: IgG1 κ

Frequently Asked Questions (FAQs)

I am unable to analyze my cells stained with eFluor Organic Dyes today. What options do I have?

Our options will depend on the samples you are analyzing.

If cell viability is not critical, you can store your stained samples at 4 degrees C or on ice overnight in the dark and analyze the following day.

For samples stained with eFluor organic fluorochromes, we recommend that cells be suspended in 100 uL of Flow Cytometry Staining Buffer (Cat. No. 00-4222) and 100 uL of eBioscience IC Fixation Buffer (Cat. No. 00-8222); samples can be incubated for up to 3 days at 4 degrees C in the dark. Alternatively, the 1-step Fix/Lyse Solution (Cat. No. 00-5333) can be used. This is a great option when working with whole blood but also works for other cell types.

Can the eFluor Organic fluorochromes be used for intracellular staining?

Yes, the eFluor Organic fluorochromes can be used for intracellular staining. The eFluor organic fluorochromes maintain bright signal and require minimal changes in compensation when fixed with eBioscience IC Fixation Buffer (Cat. No. 00-8222-49) and Permeabilization Buffer (Cat. No. 00-8333-56) or 1-step Fix/Lyse Solution (Cat. No. 00-5333-54, 00-5333-57) (as compared to live cells).

Is the eFluor 660 fluorochrome compatible with Anti-Cy5/Alexa Fluor 647 beads?

Yes, in-house studies have demonstrated that the eFluor 660 fluorochrome is recognized by Anti-Cy5/Alexa Fluor 647 beads. Side by side studies with Alexa Fluor 647 versus eFluor 660 conjugated antibodies have demonstrated comparable results.

What buffers are compatible with the eFluor Organic fluorochromes and eVolve QDots?

The eFluor Organic fluorochromes and eVolve QDots can be used with flow staining buffers containing PBS and protein.

For Research Use Only.