Revision 14
#97135
Store at -20C
Orders:
877-616-CELL (2355)
Support:
877-678-TECH (8324)
3 Trask Lane | Danvers | Massachusetts | 01923 | USA
For Research Use Only. Not for Use in Diagnostic Procedures.
Applications:
W, IP, IHC-P, IF-F, FC-FP, ChIP, C&R
Reactivity:
H M
Sensitivity:
Endogenous
MW (kDa):
62
Source/Isotype:
Rabbit IgG
UniProt ID:
#Q9JKD8
Entrez-Gene Id:
57765
Product Usage Information
The CUT&RUN dilution was determined using CUT&RUN Assay Kit #86652.
| Application | Dilution |
|---|---|
| Western Blotting | 1:1000 |
| Immunoprecipitation | 1:200 |
| Immunohistochemistry (Paraffin) | 1:100 - 1:400 |
| Immunofluorescence (Frozen) | 1:400 - 1:1600 |
| Flow Cytometry (Fixed/Permeabilized) | 1:100 - 1:400 |
| Chromatin IP | 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 #53753.
For a carrier free (BSA and azide free) version of this product see product #53753.
Specificity/Sensitivity
T-bet/TBX21 (E4I2K) Rabbit mAb recognizes endogenous levels of total T-bet/TBX21 protein. E412K non-specifically labels the submucosa and external muscularis of mouse colon by immunofluorescence.
Source / Purification
Monoclonal antibody is produced by immunizing animals with a recombinant protein fragment derived from mouse 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).
Background References
- Wilkinson, D.G. et al. (1990) Nature 343, 657-9.
- Papaioannou, V.E. and Silver, L.M. (1998) Bioessays 20, 9-19.
- Showell, C. et al. (2004) Dev Dyn 229, 201-18.
- Papaioannou, V.E. (2001) Int Rev Cytol 207, 1-70.
- Hoogaars, W.M. et al. (2007) Cell Mol Life Sci 64, 646-60.
- Baldini, A. (2004) Curr Opin Cardiol 19, 201-4.
- Abrahams, A. et al. (2010) IUBMB Life 62, 92-102.
- Rowley, M. et al. (2004) J Mammary Gland Biol Neoplasia 9, 109-18.
- Yang, X.R. et al. (2009) Nat Genet 41, 1176-8.
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
W: Western Blotting IP: Immunoprecipitation IHC-P: Immunohistochemistry (Paraffin) IF-F: Immunofluorescence (Frozen) FC-FP: Flow Cytometry (Fixed/Permeabilized) ChIP: Chromatin IP C&R: CUT&RUN
Cross-Reactivity Key
H: Human M: Mouse
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.
限制使用
除非 CST 的合法授书代表以书面形式书行明确同意，否书以下条款适用于 CST、其关书方或分书商提供的书品。 任何书充本条款或与本条款不同的客书条款和条件，除非书 CST 的合法授书代表以书面形式书独接受， 否书均被拒书，并且无效。
专品专有“专供研究使用”的专专或专似的专专声明， 且未专得美国食品和专品管理局或其他外国或国内专管机专专专任何用途的批准、准专或专可。客专不得将任何专品用于任何专断或治专目的， 或以任何不符合专专声明的方式使用专品。CST 专售或专可的专品提供专作专最专用专的客专，且专用于研专用途。将专品用于专断、专防或治专目的， 或专专售（专独或作专专成）或其他商专目的而专专专品，均需要 CST 的专独专可。客专：(a) 不得专独或与其他材料专合向任何第三方出售、专可、 出借、捐专或以其他方式专专或提供任何专品，或使用专品制造任何商专专品，(b) 不得复制、修改、逆向工程、反专专、 反专专专品或以其他方式专专专专专品的基专专专或技专，或使用专品开专任何与 CST 的专品或服专专争的专品或服专， (c) 不得更改或专除专品上的任何商专、商品名称、徽专、专利或版专声明或专专，(d) 只能根据 CST 的专品专售条款和任何适用文档使用专品 ， (e) 专遵守客专与专品一起使用的任何第三方专品或服专的任何专可、服专条款或专似专专
Orders: 877-616-CELL (2355) • [email protected] • Support: 877-678-TECH (8324) • [email protected] •
Web:
cellsignal.com
For Research Use Only. Not for Use in Diagnostic Procedures.
Revision 14
Western blot analysis of extracts from various cell lines and mouse primary CD4+ T cells using T-bet/TBX21 (E4I2K) Rabbit mAb (upper) and β-Actin (D6A8) Rabbit mAb #8457 (lower). As expected, T-bet/TBX21 protein is not detected in EL4 cells.
Western blot analysis of extracts from 293T cells, mock transfected (-) or transfected with constructs expressing Myc/DDK-tagged full-length human T-bet/TBX21 protein (hT-bet/TBX21-Myc/DDK; +) and Myc/DDK-tagged full-length mouse T-Bet/TBX21 protein (mT-bet/TBX21-Myc/DDK; +), using T-bet/TBX21 (E4I2K) Rabbit mAb (upper), DYKDDDDK Tag Antibody #2368 (middle), and β-Actin (D6A8) Rabbit mAb #8457 (lower).
Immunoprecipitation of T-bet/TBX21 protein from NK-92 cell extracts. Lane 1 is 10% input, lane 2 is Rabbit (DA1E) mAb IgG XP® Isotype Control #3900, and lane 3 is T-bet/TBX21 (E4I2K) Rabbit mAb. Western blot analysis was performed using T-bet/TBX21 (E4I2K) Rabbit mAb. Mouse Anti-rabbit IgG (Conformation Specific) (L27A9) mAb (HRP Conjugate) #5127 was used as a secondary antibody.
Orders: 877-616-CELL (2355) • [email protected] • Support: 877-678-TECH (8324) • [email protected] •
Web:
cellsignal.com
For Research Use Only. Not for Use in Diagnostic Procedures.
Revision 14
Immunohistochemical analysis of paraffin-embedded 293T cell pellets, control (left) or mouse T-bet/TBX21-transfected (right), using T-bet/TBX21 (E4I2K) Rabbit mAb.
Immunohistochemical analysis of paraffin-embedded normal human spleen using T-bet/TBX21 (E4I2K) Rabbit mAb.
Immunohistochemical analysis of paraffin-embedded human colon adenocarcinoma using T-bet/TBX21 (E4I2K) Rabbit mAb.
Orders: 877-616-CELL (2355) • [email protected] • Support: 877-678-TECH (8324) • [email protected] •
Web:
cellsignal.com
For Research Use Only. Not for Use in Diagnostic Procedures.
Revision 14
Immunohistochemical analysis of paraffin-embedded human small cell carcinoma of the salivary gland using T-bet/TBX21 (E4I2K) Rabbit mAb.
Immunohistochemical analysis of paraffin-embedded human gastric carcinoma using T-bet/TBX21 (E4I2K) Rabbit mAb.
Immunohistochemical analysis of paraffin-embedded mouse thymus using T-bet/TBX21 (E4I2K) Rabbit mAb.
Orders: 877-616-CELL (2355) • [email protected] • Support: 877-678-TECH (8324) • [email protected] •
Web:
cellsignal.com
For Research Use Only. Not for Use in Diagnostic Procedures.
Revision 14
Immunohistochemical analysis of paraffin-embedded mouse lung using T-bet/TBX21 (E4I2K) Rabbit mAb.
Immunohistochemical analysis of paraffin-embedded mouse ovary using T-bet/TBX21 (E4I2K) Rabbit mAb.
Immunohistochemical analysis of paraffin-embedded CT26.WT syngeneic tumor using T-bet/TBX21 (E4I2K) Rabbit mAb.
Orders: 877-616-CELL (2355) • [email protected] • Support: 877-678-TECH (8324) • [email protected] •
Web:
cellsignal.com
For Research Use Only. Not for Use in Diagnostic Procedures.
Revision 14
Immunohistochemical analysis of paraffin-embedded LL/2 syngeneic tumor using T-bet/TBX21 (E4I2K) Rabbit mAb.
Immunohistochemical analysis of paraffin-embedded human colon adenocarcinoma using T-bet/TBX21 (E4I2K) Rabbit mAb (left) or T-bet/TBX21 (D6N8B) XP® Rabbit mAb (right) #13232. These two antibodies detect unique, non-overlapping epitopes on human T-bet/TBX21. The similar staining patterns obtained with both antibodies help to confirm the specificity of the staining.
Immunohistochemical analysis of paraffin-embedded human B-cell non-Hodgkin lymphoma using T-bet/TBX21 (E4I2K) Rabbit mAb (left) or T-bet/TBX21 (D6N8B) XP® Rabbit mAb (right) #13232. These two antibodies detect unique, non-overlapping epitopes on human T-bet/TBX21. The similar staining patterns obtained with both antibodies help to confirm the specificity of the staining.
Orders: 877-616-CELL (2355) • [email protected] • Support: 877-678-TECH (8324) • [email protected] •
Web:
cellsignal.com
For Research Use Only. Not for Use in Diagnostic Procedures.
Revision 14
Immunohistochemical analysis of paraffin-embedded mouse spleen using T-bet/TBX21 (E4I2K) Rabbit mAb (left) compared to concentration-matched Rabbit (DA1E) mAb IgG XP® Isotype Control #3900 (right).
Confocal immunofluorescent analysis of mouse thymus using T-bet/TBX21 (E4I2K) Rabbit mAb (red), CD8α (2.43) Rat mAb (FITC Conjugate) #35467 (green), and CD45 (D3F8Q) Rabbit mAb (Alexa Fluor® 647 Conjugate) #81143 (blue pseudocolor).
Flow cytometric analysis of K-562 cells (blue, negative) and NK-92 (green, positive) using T-bet/TBX21 (E4I2K) Rabbit mAb (solid lines) or a concentration-matched Rabbit (DA1E) mAb IgG XP® Isotype Control #3900 (dashed lines). Anti-rabbit IgG (H+L), F(ab')2 Fragment (Alexa Fluor® 488 Conjugate) #4412 was used as a secondary antibody.
Orders: 877-616-CELL (2355) • [email protected] • Support: 877-678-TECH (8324) • [email protected] •
Web:
cellsignal.com
For Research Use Only. Not for Use in Diagnostic Procedures.
Revision 14
Flow cytometric analysis of EL4 cells (blue, negative) and CTLL-2 cells (green, positive) using T-bet/TBX21 (E4I2K) Rabbit mAb (solid lines) or a concentration-matched Rabbit (DA1E) mAb IgG XP® Isotype Control #3900 (dashed lines). Anti-rabbit IgG (H+L), F(ab')2 Fragment (Alexa Fluor® 488 Conjugate) #4412 was used as a secondary antibody.
Chromatin immunoprecipitations were performed with cross-linked chromatin from NK-92 cells and either T-bet/TBX21 (E4I2K) Rabbit mAb or Normal Rabbit IgG #2729 using SimpleChIP® Plus Enzymatic Chromatin IP Kit (Magnetic Beads) #9005. 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&RUN was performed with NK-92 cells and T-bet/TBX21 (E4I2K) 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, a known target gene of T-bet (see additional figure containing CUT&RUN-qPCR data).
Orders: 877-616-CELL (2355) • [email protected] • Support: 877-678-TECH (8324) • [email protected] •
Web:
cellsignal.com
For Research Use Only. Not for Use in Diagnostic Procedures.
Revision 14
CUT&RUN was performed with NK-92 cells and T-bet/TBX21 (E4I2K) 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 figures show binding across chromosome 1 (upper), including CKS1B (lower), a known target gene of T-bet (see additional figure containing CUT&RUN-qPCR data).
CUT&RUN was performed with NK-92 cells and either T-bet/TBX21 (E4I2K) 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 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.
Orders: 877-616-CELL (2355) • [email protected] • Support: 877-678-TECH (8324) • [email protected] •
Web:
cellsignal.com
For Research Use Only. Not for Use in Diagnostic Procedures.