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.

使用に関する制限

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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).