Revision 8

#5852Store at -20C

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

REACTIVITY:

H M

SENSITIVITY:

Endogenous

MW (kDa):

48

Source/Isotype:

Rabbit IgG

UniProt ID:

#P23771

Entrez-Gene Id:

2625

Product Information

Product Usage Information

For optimal ChIP results, use 5 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
Immunohistochemistry (Paraffin) 1:1600 - 1:6400
Immunofluorescence (Immunocytochemistry) 1:800 - 1:3200
Flow Cytometry (Fixed/Permeabilized) 1:200 - 1:800
Chromatin IP 1:100
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.

Specificity / Sensitivity

GATA-3 (D13C9) XP® Rabbit mAb recognizes endogenous levels of total GATA-3 protein.

Species Reactivity:

Human, Mouse

Species predicted to react based on 100% sequence homology

Monkey

Source / Purification

Monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to residues surrounding Tyr63 of human GATA-3 protein.

Background

GATA proteins comprise a group of transcription factors that are related by the presence of conserved zinc finger DNA-binding domains, which bind directly to the nucleotide sequence core element GATA (1-3). There are six vertebrate GATA proteins, designated GATA-1 to GATA-6 (3).

GATA-3 is a critical regulator of development of various systems in both mouse and human (4). GATA-3 mouse embryos die between E11 and E12 due to growth retardation and deformities in the brain and spinal cord (5). The function of GATA-3 has been extensively studied in T cell development and has recently been shown to be a downstream target of Notch in Notch-mediated differentiation of TH2 cells (6,7). It is expressed in both hematopoietic and non-hematopoietic tissues, including the kidney, skin, mammary gland, and central nervous system (8-10). Decreased expression of GATA-3 in luminal breast cancer is associated with poor clinical outcome. GATA-3 expression level may therefore be a promising prognostic biomarker (11). Haploinsufficiency of GATA-3 results in Barakat syndome in humans, a condition characterized by sensorineural deafness and renal dysplasia (12).

  1. Ko, L.J. and Engel, J.D. (1993) Mol Cell Biol 13, 4011-22.
  2. Merika, M. and Orkin, S.H. (1993) Mol Cell Biol 13, 3999-4010.
  3. Lowry, J.A. and Atchley, W.R. (2000) J Mol Evol 50, 103-15.
  4. Debacker, C. et al. (1999) Mech Dev 85, 183-7.
  5. Pandolfi, P.P. et al. (1995) Nat Genet 11, 40-4.
  6. Ho, I.C. et al. (2009) Nat Rev Immunol 9, 125-35.
  7. Amsen, D. et al. (2007) Immunity 27, 89-99.
  8. Grote, D. et al. (2008) PLoS Genet 4, e1000316.
  9. Kaufman, C.K. et al. (2003) Genes Dev 17, 2108-22.
  10. Kouros-Mehr, H. et al. (2006) Cell 127, 1041-55.
  11. Chou, J. et al. (2010) J Cell Physiol 222, 42-9.
  12. Van Esch, H. et al. (2000) Nature 406, 419-22.

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 IHC-P: Immunohistochemistry (Paraffin) IF-IC: Immunofluorescence (Immunocytochemistry) FC-FP: Flow Cytometry (Fixed/Permeabilized) ChIP: Chromatin IP 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.
XP is a registered trademark of Cell Signaling Technology, Inc.
All other trademarks are the property of their respective owners. Visit cellsignal.com/trademarks for more information.

限制使用

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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
仅供研究使用。不得用于诊断流程。
Revision 8
#5852

GATA-3 (D13C9) XP® Rabbit mAb

Western Blotting Image 1: GATA-3 (D13C9) XP® Rabbit mAb Expand Image
使用 GATA-3 (D13C9) XP® Rabbit mAb 对 CEM 和 SH-SY5Y 细胞提取物进行蛋白质印迹分析。
Immunohistochemistry Image 1: GATA-3 (D13C9) XP® Rabbit mAb Expand Image
使用 GATA-3 (D13C9) XP® Rabbit mAb（左）或 Rabbit (DA1E) mAb IgG XP® Isotype Control #3900（右），对石蜡包埋的人乳腺癌进行免疫组织化学分析。
Immunohistochemistry Image 2: GATA-3 (D13C9) XP® Rabbit mAb Expand Image
使用 GATA-3 (D13C9) XP® Rabbit mAb 对石蜡包埋的 MCF-7（左）或 HUVEC（右）细胞沉淀物进行免疫组织化学分析。
Immunofluorescence Image 1: GATA-3 (D13C9) XP® Rabbit mAb Expand Image
使用 GATA-3 (D13C9) XP® Rabbit mAb（绿色），对 MCF7（左）和 HUVE（右）细胞进行共聚焦免疫荧光分析。肌动蛋白纤丝用 DY-554 Phalloidin 进行标记（红色）。
Flow Cytometry Image 1: GATA-3 (D13C9) XP® Rabbit mAb Expand Image
使用 GATA-3 (D13C9) XP® Rabbit mAb（实线）或浓度匹配的 Rabbit (DA1E) mAb IgG XP® Isotype Control #3900（虚线）对 THP-1 细胞（蓝色，阴性）和 MCF7 细胞（绿色，阳性）进行流式细胞分析。Anti-rabbit IgG (H+L)、F(ab')2 Fragment (Alexa Fluor® 488 Conjugate) #4412 作为二抗。
Flow Cytometry Image 2: GATA-3 (D13C9) XP® Rabbit mAb Expand Image
使用 GATA-3 (D13C9) XP® Rabbit mAb 对 THP-1 细胞（蓝色） 和 MCF7 细胞（绿色）进行流式细胞分析。
Chromatin Immunoprecipitation Image 1: GATA-3 (D13C9) XP® Rabbit mAb Expand Image
使用 SimpleChIP® Enzymatic Chromatin IP Kit (Magnetic Beads) #9003，对 NK-92 细胞中提取的交联染色质，在加入 GATA-3 (D13C9) XP® Rabbit mAb 或 Normal Rabbit IgG #2729 后进行染色质免疫沉淀。使用 Human CCL4 Promoter Primers、SimpleChIP® Human IFN-γ Promoter Primers #13051 和 SimpleChIP® Human α Satellite Repeat Primers #4486 并通过实时 PCR 对富集的 DNA 进行定量分析。将每份样品中免疫沉淀的 DNA 的量表现为相对于所输入染色质总量（等于 1）的信号。
CUT and RUN Image 1: GATA-3 (D13C9) XP® Rabbit mAb Expand Image
使用 CUT&RUN Assay Kit #86652 对 NK-92 细胞和 GATA-3 (D13C9) XP® Rabbit mAb 或 GATA-3 (E2N1Y) Mouse mAb #96098 进行 CUT&RUN 检测。使用 SimpleChIP® ChIP-seq DNA Library Prep Kit for Illumina® #56795 制备 DNA 文库。该图显示了 GATA-3 的已知靶基因 SH3RF3 内结合（参见包含 CUT&RUN-qPCR 数据的其他结果图）。
CUT and RUN Image 2: GATA-3 (D13C9) XP® Rabbit mAb Expand Image
使用 CUT&RUN Assay Kit #86652 对 NK-92 细胞和 GATA-3 (D13C9) XP® Rabbit mAb 或 GATA-3 (E2N1Y) Mouse mAb #96098 进行 CUT&RUN 检测。使用 SimpleChIP® ChIP-seq DNA Library Prep Kit for Illumina® #56795 制备 DNA 文库。该图显示了在染色体 2（上图）内的结合，包括 GATA-3 的已知靶标基因 SH3RF3（下图）（参见包含 ChIP-qPCR 数据的其他结果图）。
CUT and RUN Image 3: GATA-3 (D13C9) XP® Rabbit mAb Expand Image
使用 CUT&RUN Assay Kit #86652 对 NK-92 细胞和 GATA-3 (D13C9) XP® Rabbit mAb 或 Rabbit (DA1E) mAb IgG XP® Isotype Control (CUT&RUN) #66362 进行 CUT&RUN 检测。使用 human RBM15 promoter primer、human SH3RF3 intron 1 primer 和 SimpleChIP® Human α Satellite Repeat Primers #4486，通过实时 PCR 对富集的 DNA 进行定量分析。将每份样品中免疫沉淀的 DNA 的量表现为相对于所输入染色质总量（等于 1）的信号。
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Orders: 877-616-CELL (2355)•[email protected]• Support: 877-678-TECH (8324)•[email protected]• Web: cellsignal.com
仅供研究使用。不得用于诊断流程。