Revision 5

#96098Store 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, C&R

REACTIVITY:

H

SENSITIVITY:

Endogenous

MW (kDa):

50

Source/Isotype:

Mouse IgG1

UniProt ID:

#P23771

Entrez-Gene Id:

2625

Product Information

Product Usage Information

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

For a carrier free (BSA and azide free) version of this product see product #57847.

Specificity / Sensitivity

GATA-3 (E2N1Y) Mouse mAb recognizes endogenous levels of total GATA-3 protein.

Species Reactivity:

Human

Source / Purification

Monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to residues near the amino terminus 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) 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 5
#96098

GATA-3 (E2N1Y) Mouse mAb

Western Blotting Image 1: GATA-3 (E2N1Y) Mouse mAb Expand Image
Western blot analysis of extracts from CCRF-CEM and RD cells using GATA-3 (E2N1Y) Mouse mAb (upper) and β-Actin (D6A8) Rabbit mAb #8457 (lower). As expected RD cells show low to negative expression of GATA-3.
Immunohistochemistry Image 1: GATA-3 (E2N1Y) Mouse mAb Expand Image
Immunohistochemical analysis of paraffin-embedded human ductal breast carcinoma (left, positive) or colon carcinoma (right, negative) using GATA-3 (E2N1Y) Mouse mAb.
Immunohistochemistry Image 2: GATA-3 (E2N1Y) Mouse mAb Expand Image
Immunohistochemical analysis of paraffin-embedded MCF7 cell pellet (left, positive) or HUVEC cell pellet (right, negative) using GATA-3 (E2N1Y) Mouse mAb.
Immunohistochemistry Image 3: GATA-3 (E2N1Y) Mouse mAb Expand Image
Immunohistochemical analysis of paraffin-embedded human urothelial carcinoma using GATA-3 (E2N1Y) Mouse mAb.
Immunofluorescence Image 1: GATA-3 (E2N1Y) Mouse mAb Expand Image
Confocal immunofluorescent analysis of MCF7 cells (left, positive) and Hep G2 cells (right, negative) using GATA-3 (E2N1Y) Rabbit mAb (green). Actin filaments were labeled with DyLight 554 Phalloidin #13054 (red).
Flow Cytometry Image 1: GATA-3 (E2N1Y) Mouse mAb Expand Image
Flow cytometric analysis of THP-1 cells (blue) and MCF7 cells (green) using GATA3 (E2N1Y) Mouse mAb (solid lines) or concentration-matched Mouse (G3A1) mAb IgG Isotype control #5415 (dashed lines). Anti-mouse IgG (H+L), F(ab')2 Fragment (Alexa Fluor® 488 Conjugate) #4408 was used as a secondary antibody.
CUT and RUN Image 1: GATA-3 (E2N1Y) Mouse mAb Expand Image
CUT&RUN was performed with NK-92 cells and either GATA-3 (E2N1Y) Mouse mAb or GATA-3 (D13C9) XP® Rabbit mAb #5852, 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 SH3RF3 gene.
CUT and RUN Image 2: GATA-3 (E2N1Y) Mouse mAb Expand Image
CUT&RUN was performed with NK-92 cells and either GATA-3 (E2N1Y) Mouse mAb or GATA-3 (D13C9) XP® Rabbit mAb #5852, 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 2 (upper), including SH3RF3 gene (lower).
CUT and RUN Image 3: GATA-3 (E2N1Y) Mouse mAb Expand Image
CUT&RUN was performed with NK-92 cells and either GATA-3 (E2N1Y) Mouse 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 RBM15 promoter primers 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.