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.

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

Revision 6
#5851

GATA-6 (D61E4) XP® Rabbit mAb

Western Blotting Image 1: GATA-6 (D61E4) XP® Rabbit mAb Expand Image
使用 GATA-6 (D61E4) XP® Rabbit mAb 对 Huh7 细胞和 293 细胞提取物进行蛋白质印迹分析。
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
使用 GATA-6 (D61E4) XP® Rabbit mAb(绿色)对 KM12 细胞(左)和 SK-OV-3 细胞(右)进行共聚焦免疫荧光分析。使用 DY-554 phalloidin(红色)标记肌动蛋白纤丝。
Flow Cytometry Image 1: GATA-6 (D61E4) XP® Rabbit mAb Expand Image
使用 GATA-6 (D61E4) XP® Rabbit mAb 对 SKOV3 细胞(蓝色)和 HUH-7 细胞(绿色)进行流式细胞分析。使用 Anti-rabbit IgG (H+L), F(ab')2 Fragment (Alexa Fluor 488 Conjugate) #4412 作为二抗。
Chromatin Immunoprecipitation Image 1: GATA-6 (D61E4) XP® Rabbit mAb Expand Image
使用 SimpleChIP® Plus Enzymatic Chromatin IP Kit (Magnetic Beads) #9005,对 Caco-2 细胞中提取的交联染色质,在加入 GATA-6 (D61E4) XP® Rabbit mAb 后进行染色质免疫沉淀。使用 DNA Library Prep Kit for Illumina® (ChIP-seq, CUT&RUN) #56795 制备 DNA 库。本图显示在 FGFR2(GATA-6 的一个已知靶标基因)内的结合作用(参见包含 ChIP-qPCR 数据的其他图)。如需了解其他 ChIP-seq 情况,请下载产品说明书。
Chromatin Immunoprecipitation Image 2: GATA-6 (D61E4) XP® Rabbit mAb Expand Image
使用 SimpleChIP® Plus Enzymatic Chromatin IP Kit (Magnetic Beads) #9005,对 Caco-2 细胞中提取的交联染色质,在加入 GATA-6 (D61E4) XP® Rabbit mAb 后进行染色质免疫沉淀。使用 DNA Library Prep Kit for Illumina® (ChIP-seq, CUT&RUN) #56795 制备 DNA 库。该结果图显示在染色体 10(上图)内的结合,包括 GATA-6 的已知靶基因 FGFR2(下图)(参见包含 ChIP-qPCR 数据的其他结果图)。
Chromatin Immunoprecipitation Image 3: GATA-6 (D61E4) XP® Rabbit mAb Expand Image
使用 SimpleChIP® Enzymatic Chromatin IP Kit (Magnetic Beads) #9003,对 Caco-2 细胞中提取的交联染色质,在加入 GATA-6 (D61E4) XP® Rabbit mAb 或 Normal Rabbit IgG #2729 后进行染色质免疫沉淀。使用 SimpleChIP® Human FDPS Promoter Primers #13840、人 FGFR2 启动子引物和 SimpleChIP® Human α Satellite Repeat Primers #4486,通过实时 PCR 对富集的 DNA 进行定量。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-6 (D61E4) XP® Rabbit mAb Expand Image
使用 CUT&RUN Assay Kit #86652,Caco-2 细胞和 GATA-6 (D61E4) XP® Rabbit mAb 进行 CUT&RUN。使用 DNA Library Prep Kit for Illumina® (ChIP-seq, CUT&RUN) #56795 制备 DNA 库。结果图显示在 FGFR2 基因内结合。
CUT and RUN Image 2: GATA-6 (D61E4) XP® Rabbit mAb Expand Image
使用 CUT&RUN Assay Kit #86652,Caco-2 细胞和 GATA-6 (D61E4) XP® Rabbit mAb 进行 CUT&RUN。使用 DNA Library Prep Kit for Illumina® (ChIP-seq, CUT&RUN) #56795 制备 DNA 库。结果图显示在染色体 10(上图)内的结合,包括 FGFR2 基因(下图)。
CUT and RUN Image 3: GATA-6 (D61E4) XP® Rabbit mAb Expand Image
使用 CUT&RUN Assay Kit #86652,通过 Caco-2 细胞和 GATA-6 (D61E4) XP® Rabbit mAb 或 Rabbit (DA1E) mAb IgG XP® Isotype Control (CUT&RUN) #66362 进行 CUT&RUN。使用 人源 HNMT 上游引物和 SimpleChIP® Human α Satellite Repeat Primers #4486 进行实时 PCR 来对富集的 DNA 进行定量。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 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).