Revision 6

#5851Store 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, ChIP-seq, C&R, C&T

REACTIVITY:

H M

SENSITIVITY:

Endogenous

MW (kDa):

55

Source/Isotype:

Rabbit IgG

UniProt ID:

#Q92908

Entrez-Gene Id:

2627

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.

The CUT&Tag dilution was determined using CUT&Tag Assay Kit #77552.

Application Dilution
Western Blotting 1:1000
Immunohistochemistry (Paraffin) 1:100 - 1:400
Immunofluorescence (Immunocytochemistry) 1:1600
Flow Cytometry (Fixed/Permeabilized) 1:800
Chromatin IP 1:50
Chromatin IP-seq 1:50
CUT&RUN 1:50
CUT&Tag 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 #48888.

Specificity / Sensitivity

GATA-6 (D61E4) XP® Rabbit mAb recognizes endogenous levels of total GATA-6 protein. Non-specific cytoplasmic staining of limited immune cells in mouse small intestine, thymus and spleen was observed by immunohistochemistry.

Species Reactivity:

Human, Mouse

Species predicted to react based on 100% sequence homology

Mouse, Rat, Dog, Pig

Source / Purification

Monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to residues near the amino terminus of human GATA-6 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-6 plays a critical role in endoderm development (4). It is essential for development of the heart, gut, and other organs (5,6). Knock out of GATA-6 is embryonic lethal due to defects in formation of the heart tube and a failure to develop extraembryonic endoderm (4). Loss of expression, or loss of nuclear localization of GATA-6 is apparent in a large number of ovarian tumors (7).

  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. Cai, K.Q. et al. (2008) Dev Dyn 237, 2820-9.
  5. Charron, F. and Nemer, M. (1999) Semin Cell Dev Biol 10, 85-91.
  6. Haveri, H. et al. (2008) BMC Gastroenterol 8, 9.
  7. Caslini, C. et al. (2006) Oncogene 25, 5446-61.

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 ChIP-seq: Chromatin IP-seq C&R: CUT&RUN C&T: CUT&Tag

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.

Limited Uses

Except as otherwise expressly agreed in a writing signed by a legally authorized representative of CST, the following terms apply to Products provided by CST, its affiliates or its distributors. Any Customer's terms and conditions that are in addition to, or different from, those contained herein, unless separately accepted in writing by a legally authorized representative of CST, are rejected and are of no force or effect.

Products are labeled with For Research Use Only or a similar labeling statement and have not been approved, cleared, or licensed by the FDA or other regulatory foreign or domestic entity, for any purpose. Customer shall not use any Product for any diagnostic or therapeutic purpose, or otherwise in any manner that conflicts with its labeling statement. Products sold or licensed by CST are provided for Customer as the end-user and solely for research and development uses. Any use of Product for diagnostic, prophylactic or therapeutic purposes, or any purchase of Product for resale (alone or as a component) or other commercial purpose, requires a separate license from CST. Customer shall (a) not sell, license, loan, donate or otherwise transfer or make available any Product to any third party, whether alone or in combination with other materials, or use the Products to manufacture any commercial products, (b) not copy, modify, reverse engineer, decompile, disassemble or otherwise attempt to discover the underlying structure or technology of the Products, or use the Products for the purpose of developing any products or services that would compete with CST products or services, (c) not alter or remove from the Products any trademarks, trade names, logos, patent or copyright notices or markings, (d) use the Products solely in accordance with CST Product Terms of Sale and any applicable documentation, and (e) comply with any license, terms of service or similar agreement with respect to any third party products or services used by Customer in connection with the Products.

Revision 6
#5851

GATA-6 (D61E4) XP® Rabbit mAb

Western Blotting Image 1: GATA-6 (D61E4) XP® Rabbit mAb Expand Image
Western blot analysis of extracts from Huh7 and 293 cells using GATA-6 (D61E4) XP® Rabbit mAb.
Immunohistochemistry Image 1: GATA-6 (D61E4) XP® Rabbit mAb Expand Image
Immunohistochemical analysis of paraffin-embedded human esophageal carcinoma using GATA-6 (D61E4) XP® Rabbit mAb.
Immunohistochemistry Image 2: GATA-6 (D61E4) XP® Rabbit mAb Expand Image
Immunohistochemical analysis of paraffin-embedded human granulosa cell tumor of the ovary using GATA-6 (D61E4) XP® Rabbit mAb.
Immunohistochemistry Image 3: GATA-6 (D61E4) XP® Rabbit mAb Expand Image
Immunohistochemical analysis of paraffin-embedded human gastric carcinoma using GATA-6 (D61E4) XP® Rabbit mAb.
Immunohistochemistry Image 4: GATA-6 (D61E4) XP® Rabbit mAb Expand Image
Immunohistochemical analysis of paraffin-embedded human serous papillary carcinoma of the ovary using GATA-6 (D61E4) XP® Rabbit mAb.
Immunohistochemistry Image 5: GATA-6 (D61E4) XP® Rabbit mAb Expand Image
Immunohistochemical analysis of paraffin-embedded human squamous cell lung carcinoma using GATA-6 (D61E4) XP® Rabbit mAb.
Immunohistochemistry Image 6: GATA-6 (D61E4) XP® Rabbit mAb Expand Image
Immunohistochemical analysis of paraffin-embedded human lung adenocarcinoma using GATA-6 (D61E4) XP® Rabbit mAb.
Immunohistochemistry Image 7: GATA-6 (D61E4) XP® Rabbit mAb Expand Image
Immunohistochemical analysis of paraffin-embedded human colon carcinoma using GATA-6 (D61E4) XP® Rabbit mAb.
Immunohistochemistry Image 8: GATA-6 (D61E4) XP® Rabbit mAb Expand Image
Immunohistochemical analysis of paraffin-embedded normal human stomach using GATA-6 (D61E4) XP® Rabbit mAb.
Immunohistochemistry Image 9: GATA-6 (D61E4) XP® Rabbit mAb Expand Image
Immunohistochemical analysis of paraffin-embedded normal human small intestine using GATA-6 (D61E4) XP® Rabbit mAb.
Immunohistochemistry Image 10: GATA-6 (D61E4) XP® Rabbit mAb Expand Image
Immunohistochemical analysis of paraffin-embedded normal human pancreas using GATA-6 (D61E4) XP® Rabbit mAb.
Immunohistochemistry Image 11: GATA-6 (D61E4) XP® Rabbit mAb Expand Image
Immunohistochemical analysis of paraffin-embedded normal human ovary using GATA-6 (D61E4) XP® Rabbit mAb.
Immunohistochemistry Image 12: GATA-6 (D61E4) XP® Rabbit mAb Expand Image
>Immunohistochemical analysis of paraffin-embedded normal human lung using GATA-6 (D61E4) XP® Rabbit mAb.
Immunohistochemistry Image 13: GATA-6 (D61E4) XP® Rabbit mAb Expand Image
Immunohistochemical analysis of paraffin-embedded normal human heart using GATA-6 (D61E4) XP® Rabbit mAb.
Immunohistochemistry Image 14: GATA-6 (D61E4) XP® Rabbit mAb Expand Image
Immunohistochemical analysis of paraffin-embedded human appendix using GATA-6 (D61E4) XP® Rabbit mAb.
Immunohistochemistry Image 15: GATA-6 (D61E4) XP® Rabbit mAb Expand Image
Immunohistochemical analysis of paraffin-embedded normal human adrenal gland using GATA-6 (D61E4) XP ® Rabbit mAb.
Immunohistochemistry Image 16: GATA-6 (D61E4) XP® Rabbit mAb Expand Image
Immunohistochemical analysis of paraffin-embedded LL/2 syngeneic tumor using GATA-6 (D61E4) XP® Rabbit mAb.
Immunohistochemistry Image 17: GATA-6 (D61E4) XP® Rabbit mAb Expand Image
Immunohistochemical analysis of paraffin-embedded 4T1 syngeneic tumor using GATA-6 (D61E4) XP® Rabbit mAb.
Immunohistochemistry Image 18: GATA-6 (D61E4) XP® Rabbit mAb Expand Image
Immunohistochemical analysis of paraffin-embedded mouse testis using GATA-6 (D61E4) XP® Rabbit mAb.
Immunohistochemistry Image 19: GATA-6 (D61E4) XP® Rabbit mAb Expand Image
Immunohistochemical analysis of paraffin-embedded mouse ovary using GATA-6 (D61E4) XP® Rabbit mAb.
Immunohistochemistry Image 20: GATA-6 (D61E4) XP® Rabbit mAb Expand Image
Immunohistochemical analysis of paraffin-embedded mouse colon using GATA-6 (D61E4) XP® Rabbit mAb
Immunohistochemistry Image 21: GATA-6 (D61E4) XP® Rabbit mAb Expand Image
Immunohistochemical analysis of paraffin-embedded mouse small intestine using GATA-6 (D61E4) XP® Rabbit mAb.
Immunohistochemistry Image 22: GATA-6 (D61E4) XP® Rabbit mAb Expand Image
Immunohistochemical analysis of paraffin-embedded mouse brain using GATA-6 (D61E4) XP® Rabbit mAb.
Immunohistochemistry Image 23: GATA-6 (D61E4) XP® Rabbit mAb Expand Image
Immunohistochemical analysis of paraffin-embedded normal human colon using GATA-6 (D61E4) XP® Rabbit mAb (left) compared to concentration-matched Rabbit (DA1E) mAb IgG XP® Isotype Control #3900 (right).
Immunohistochemistry Image 24: GATA-6 (D61E4) XP® Rabbit mAb Expand Image

Immunohistochemical analysis of paraffin-embedded HuH-7 cell pellet (left, positive) and SUP-B15 cell pellet (right, negative) using GATA-6 (D61E4) XP® Rabbit mAb.

Immunofluorescence Image 1: GATA-6 (D61E4) XP® Rabbit mAb Expand Image
Confocal immunofluorescent analysis of KM12 (left) and SK-OV-3 cells (right) using GATA-6 (D61E4) XP® Rabbit mAb (green). Actin filaments were labeled with DY-554 phalloidin (red).
Flow Cytometry Image 1: GATA-6 (D61E4) XP® Rabbit mAb Expand Image
Flow cytometric analysis of SKOV3 cells (blue) and HUH-7 cells (green) using GATA-6 (D61E4) XP® Rabbit mAb. Anti-rabbit IgG (H+L), F(ab')2 Fragment (Alexa Fluor 488 Conjugate) #4412 was used as a secondary antibody.
Chromatin Immunoprecipitation Image 1: GATA-6 (D61E4) XP® Rabbit mAb Expand Image
Chromatin immunoprecipitations were performed with cross-linked chromatin from Caco-2 cells and GATA-6 (D61E4) XP® Rabbit mAb, using SimpleChIP® Plus Enzymatic Chromatin IP Kit (Magnetic Beads) #9005. DNA Libraries were prepared using DNA Library Prep Kit for Illumina® (ChIP-seq, CUT&RUN) #56795. The figure shows binding across FGFR2, a known target gene of GATA-6 (see additional figure containing ChIP-qPCR data). For additional ChIP-seq tracks, please download the product data sheet.
Chromatin Immunoprecipitation Image 2: GATA-6 (D61E4) XP® Rabbit mAb Expand Image
Chromatin immunoprecipitations were performed with cross-linked chromatin from Caco-2 cells and GATA-6 (D61E4) XP® Rabbit mAb, using SimpleChIP® Plus Enzymatic Chromatin IP Kit (Magnetic Beads) #9005. DNA Libraries were prepared using DNA Library Prep Kit for Illumina® (ChIP-seq, CUT&RUN) #56795. The figure shows binding across chromosome 10 (upper), including FGFR2 (lower), a known target gene of GATA-6 (see additional figure containing ChIP-qPCR data).
Chromatin Immunoprecipitation Image 3: GATA-6 (D61E4) XP® Rabbit mAb Expand Image
Chromatin immunoprecipitations were performed with cross-linked chromatin from Caco-2 cells and either GATA-6 (D61E4) 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 SimpleChIP® Human FDPS Promoter Primers #13840, human FGFR2 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.
CUT and RUN Image 1: GATA-6 (D61E4) XP® Rabbit mAb Expand Image
CUT&RUN was performed with Caco-2 cells and GATA-6 (D61E4) 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 figure shows binding across FGFR2 gene.
CUT and RUN Image 2: GATA-6 (D61E4) XP® Rabbit mAb Expand Image
CUT&RUN was performed with Caco-2 cells and GATA-6 (D61E4) 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 10 (upper), including FGFR2 gene (lower).
CUT and RUN Image 3: GATA-6 (D61E4) XP® Rabbit mAb Expand Image
CUT&RUN was performed with Caco-2 cells and either GATA-6 (D61E4) 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 HNMT upstream primer, 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 Tag Image 1: GATA-6 (D61E4) XP® Rabbit mAb Expand Image
CUT&Tag was performed with Caco-2 cells and GATA-6 (D61E4) XP® Rabbit mAb, using CUT&Tag Assay Kit #77552. DNA library was prepared using CUT&Tag Dual Index Primers and PCR Master Mix for Illumina Systems #47415. The figure shows binding across HNMT, a known target gene of GATA-6 (see our CUT&RUN-qPCR figure).
CUT and Tag Image 2: GATA-6 (D61E4) XP® Rabbit mAb Expand Image
CUT&Tag was performed with Caco-2 cells and GATA-6 (D61E4) XP® Rabbit mAb, using CUT&Tag Assay Kit #77552. DNA library was prepared using CUT&Tag Dual Index Primers and PCR Master Mix for Illumina Systems #47415. The figures show binding across chromosome 2 (upper), including HNMT (lower), a known target gene of GATA-6 (see our CUT&RUN-qPCR figure).