Revision 8
Cell Signaling Technology

Orders: 877-616-CELL (2355) [email protected]

Support: 877-678-TECH (8324)

Web: [email protected] cellsignal.com

3 Trask LaneDanversMassachusetts01923USA
For Research Use Only. Not for Use in Diagnostic Procedures.
Applications:

WB, IP, IHC-P, IF-IC, FC-FP, ChIP, ChIP-seq, C&R

REACTIVITY:

H

SENSITIVITY:

Endogenous

MW (kDa):

58-68

Source/Isotype:

Rabbit IgG

UniProt ID:

#Q9UL17

Entrez-Gene Id:

30009

Product Information

Product Usage Information

For optimal ChIP and ChIP-seq results, use 10 μl of antibody and 10 μg of chromatin (approximately 4 x 106 cells) per IP. This antibody has been validated using SimpleChIP® Enzymatic Chromatin IP Kits.

The CUT&RUN dilution was determined using CUT&RUN Assay Kit #86652.
Application Dilution
Western Blotting 1:1000
Immunoprecipitation 1:100
Immunohistochemistry (Paraffin) 1:1600
Immunofluorescence (Immunocytochemistry) 1:400
Flow Cytometry (Fixed/Permeabilized) 1:100 - 1:400
Chromatin IP 1:50
Chromatin IP-seq 1:50
CUT&RUN 1:50

Storage

Supplied in 10 mM sodium HEPES (pH 7.5), 150 mM NaCl, 100 µg/ml BSA, 50% glycerol and less than 0.02% sodium azide. Store at –20°C. Do not aliquot the antibody.

For a carrier free (BSA and azide free) version of this product see product #27112.

Specificity / Sensitivity

T-bet/TBX21 (D6N8B) XP® Rabbit mAb recognizes endogenous levels of total T-bet/TBX21 protein.

Species Reactivity:

Human

Source / Purification

Monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to residues surrounding Gly465 of human T-bet/TBX21 protein.

Background

The T-box gene family consists of transcription factors characterized by a related DNA-binding domain (T-box) of approximately 200 amino acids (1,2). The T-box genes exhibit diverse temporal and spatial patterns in the developing embryo. Studies have demonstrated members of this family play crucial roles during embryogenesis in a wide range of organisms by regulating cell fate decisions to establish the early body plan and to regulate later processes underlying organogenesis (3-5). Mutations in T-box genes are associated with many developmental defects (6). Recent studies also indicate potential roles in cancer by members of the T-box family (7-9).

T-bet, also as known as TBX21, plays a critical role in development and maintenance of type 1 helper T (Th1) and T-bet deficient mice display impaired Th1 differentiation (10,11).

  1. Wilkinson, D.G. et al. (1990) Nature 343, 657-9.
  2. Papaioannou, V.E. and Silver, L.M. (1998) Bioessays 20, 9-19.
  3. Showell, C. et al. (2004) Dev Dyn 229, 201-18.
  4. Papaioannou, V.E. (2001) Int Rev Cytol 207, 1-70.
  5. Hoogaars, W.M. et al. (2007) Cell Mol Life Sci 64, 646-60.
  6. Baldini, A. (2004) Curr Opin Cardiol 19, 201-4.
  7. Abrahams, A. et al. (2010) IUBMB Life 62, 92-102.
  8. Rowley, M. et al. (2004) J Mammary Gland Biol Neoplasia 9, 109-18.
  9. Yang, X.R. et al. (2009) Nat Genet 41, 1176-8.
  10. Ho, I.C. and Glimcher, L.H. (2002) Cell 109 Suppl, S109-20.
  11. Peng, S.L. (2006) Cell Mol Immunol 3, 87-95.

Species Reactivity

Species reactivity is determined by testing in at least one approved application (e.g., western blot).

Western Blot Buffer

IMPORTANT: For western blots, incubate membrane with diluted primary antibody in 5% w/v BSA, 1X TBS, 0.1% Tween® 20 at 4°C with gentle shaking, overnight.

Applications Key

WB: Western Blotting IP: Immunoprecipitation IHC-P: Immunohistochemistry (Paraffin) IF-IC: Immunofluorescence (Immunocytochemistry) FC-FP: Flow Cytometry (Fixed/Permeabilized) ChIP: Chromatin IP ChIP-seq: Chromatin IP-seq C&R: CUT&RUN

Cross-Reactivity Key

H: human M: mouse R: rat Hm: hamster Mk: monkey Vir: virus Mi: mink C: chicken Dm: D. melanogaster X: Xenopus Z: zebrafish B: bovine Dg: dog Pg: pig Sc: S. cerevisiae Ce: C. elegans Hr: horse GP: Guinea Pig Rab: rabbit All: all species expected

Trademarks and Patents

Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc.
Alexa Fluor is a registered trademark of Life Technologies Corporation.
All other trademarks are the property of their respective owners. Visit cellsignal.com/trademarks for more information.

使用に関する制限

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Revision 8
#13232

T-bet/TBX21 (D6N8B) XP® Rabbit mAb

Western Blotting Image 1: T-bet/TBX21 (D6N8B) XP® Rabbit mAb Expand Image
Western blot analysis of extracts from NK-92 and K-562 cells using T-bet/TBX21 (D6N8B) XP® Rabbit mAb (upper) and β-Actin (D6A8) Rabbit mAb #8457 (lower).
No image available
Immunohistochemistry Image 1: T-bet/TBX21 (D6N8B) XP® Rabbit mAb Expand Image
Immunohistochemical analysis of paraffin-embedded human breast carcinoma using T-bet/TBX21 (D6N8B) XP® Rabbit mAb.
Immunohistochemistry Image 2: T-bet/TBX21 (D6N8B) XP® Rabbit mAb Expand Image
Immunohistochemical analysis of paraffin-embedded MJ [G11] cell pellet (left, positive) or K-562 cell pellet (right, negative) using T-bet/TBX21 (D6N8B) XP® Rabbit mAb.
Immunohistochemistry Image 3: T-bet/TBX21 (D6N8B) XP® Rabbit mAb Expand Image
Immunohistochemical analysis of paraffin-embedded Non-Hodgkin B-cell lymphoma using T-bet/TBX21 (D6N8B) XP® Rabbit mAb.
Immunohistochemistry Image 4: T-bet/TBX21 (D6N8B) XP® Rabbit mAb Expand Image
Immunohistochemical analysis of paraffin-embedded human tonsil using T-bet/TBX21 (D6N8B) XP® Rabbit mAb in the presence of control peptide (left) or antigen-specific peptide (right).
Immunofluorescence Image 1: T-bet/TBX21 (D6N8B) XP® Rabbit mAb Expand Image
Immunofluorescent analysis of NK-92 (positive, left) and K-562 (negative, right) cells using T-bet/TBX21 XP® (D6N8B) Rabbit mAb (green). Blue pseudocolor = DRAQ5® #4084 (fluorescent DNA dye).
Flow Cytometry Image 1: T-bet/TBX21 (D6N8B) XP® Rabbit mAb Expand Image
Flow cytometric analysis of human PBMCs using T-bet/TBX21 (D6N8B) XP® Rabbit mAb co-stained with either CD56, CD3 or CD19. CD56+ NK cells and a subset of CD3+ T cells are both distinctly positive for T-bet/TBX21 whereas CD19+ B cells are negative. Anti-rabbit IgG (H+L), F(ab')2 Fragment (Alexa Fluor® 647 Conjugate) #4414 was used as a secondary antibody for the T-Bet/TBX21 primary.
Chromatin Immunoprecipitation Image 1: T-bet/TBX21 (D6N8B) XP® Rabbit mAb Expand Image
Chromatin immunoprecipitations were performed with cross-linked chromatin from NK-92 cells and T-bet/TBX21 (D6N8B) XP® Rabbit mAb, using SimpleChIP® Plus Enzymatic Chromatin IP Kit (Magnetic Beads) #9005. DNA Libraries were prepared using SimpleChIP® ChIP-seq DNA Library Prep Kit for Illumina® #56795. The figure shows binding across CCL4, a known target gene of T-bet/TBX21 (see additional figure containing ChIP-qPCR data).
Chromatin Immunoprecipitation Image 2: T-bet/TBX21 (D6N8B) XP® Rabbit mAb Expand Image
Chromatin immunoprecipitations were performed with cross-linked chromatin from 4 x 106 NK-92 cells and T-bet/TBX21 (D6N8B) XP® Rabbit mAb, using SimpleChIP® Plus Enzymatic Chromatin IP Kit (Magnetic Beads) #9005. DNA Libraries were prepared using SimpleChIP® ChIP-seq DNA Library Prep Kit for Illumina® #56795. The figure shows binding across chromosome 17 (upper), including CCL4 (lower), a known target gene of T-bet/TBX21 (see additional figure containing ChIP-qPCR data).
Chromatin Immunoprecipitation Image 3: T-bet/TBX21 (D6N8B) XP® Rabbit mAb Expand Image
Chromatin immunoprecipitations were performed with cross-linked chromatin from NK-92 cells and either T-bet/TBX21 (D6N8B) XP® Rabbit mAb or Normal Rabbit IgG #2729 using SimpleChIP® Enzymatic Chromatin IP Kit (Magnetic Beads) #9003. The enriched DNA was quantified by real-time PCR using human CCL4 promoter primers, SimpleChIP® Human IFN-γ Promoter Primers #13051, and SimpleChIP® Human α Satellite Repeat Primers #4486. The amount of immunoprecipitated DNA in each sample is represented as signal relative to the total amount of input chromatin, which is equivalent to one.
CUT and RUN Image 1: T-bet/TBX21 (D6N8B) XP® Rabbit mAb Expand Image
CUT&RUN was performed with NK-92 cells and T-bet/TBX21 (D6N8B) XP® Rabbit mAb, using CUT&RUN Assay Kit #86652. DNA library was prepared using DNA Library Prep Kit for Illumina (ChIP-seq, CUT&RUN) #56795. The figure shows binding across CKS1B gene, a known target gene of T-bet (see additional figure containing CUT&RUN-qPCR data).
CUT and RUN Image 2: T-bet/TBX21 (D6N8B) XP® Rabbit mAb Expand Image
CUT&RUN was performed with NK-92 cells and T-bet/TBX21 (D6N8B) XP® Rabbit mAb, using CUT&RUN Assay Kit #86652. DNA Libraries were prepared using DNA Library Prep Kit for Illumina (ChIP-seq, CUT&RUN) #56795. The figures show binding across chromosome 1 (upper), including CKS1B gene (lower), a known target gene of T-bet (see additional figure containing CUT&RUN-qPCR data).
CUT and RUN Image 3: T-bet/TBX21 (D6N8B) XP® Rabbit mAb Expand Image
CUT&RUN was performed with NK-92 cells and T-bet/TBX21 (D6N8B) XP® Rabbit mAb or Rabbit (DA1E) mAb IgG XP® Isotype Control (CUT&RUN) #66362, using CUT&RUN Assay Kit #86652. The enriched DNA was quantified by real-time PCR using human GTPbP8 intron 1 primers, human CKS1B exon 1 primers, and human KC6 upstream primers. The amount of immunoprecipitated DNA in each sample is represented as signal relative to the total amount of input chromatin, which is equivalent to one.