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Anti ets2

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Anti-ETS2 is a laboratory reagent used to detect and quantify the ETS2 protein in biological samples. ETS2 is a transcription factor that plays a role in various cellular processes. Anti-ETS2 can be used in techniques such as Western blotting, immunohistochemistry, and ELISA to analyze the expression and localization of ETS2 in different cell types and tissues.

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Lab products found in correlation

Anti gfp, by Thermo Fisher Scientific (4 mentions) Ez chip kit, by Merck Group (2 mentions) Anti lamin a c, by Cell Signaling Technology (2 mentions) Trans blot turbo blotting system, by Bio-Rad (2 mentions) Criterion gel, by Bio-Rad (2 mentions) Odyssey clx imaging system, by LI COR (2 mentions) Anti myc, by Cell Signaling Technology (2 mentions) Anti nfatc1, by Thermo Fisher Scientific (2 mentions) Anti rcan1, by Merck Group (2 mentions) Anti mkp3, by Santa Cruz Biotechnology (2 mentions) Anti erk1 2, by Cell Signaling Technology (2 mentions) Anti flag, by Merck Group (2 mentions) Anti gapdh, by Biosynth (2 mentions) Anti kras, by Cell Signaling Technology (1 mentions) Anti lyn, by Cell Signaling Technology (1 mentions) Anti actb β actin, by Merck Group (1 mentions) Anti plk1, by Cell Signaling Technology (1 mentions) Anti lc3b, by Cell Signaling Technology (1 mentions) A32961, by Thermo Fisher Scientific (1 mentions) Anti phospho erk1 2, by Cell Signaling Technology (1 mentions)

7 protocols using anti ets2

1

Western Blot Quantification Protocol

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Immunoblotting was performed as previously reported34 (link),77 (link). Antibodies used in this study include: anti-LC3B (Cell Signaling Technology®, 1:1000), anti-ACTB (β-actin) (Sigma-Aldrich, 1:5000) anti-KRAS, anti-LYN (Cell Signaling Technology®, 1:1000), anti-HCLS1 (Cell Signaling Technology®, 1:1000), anti-PLK1 (Cell Signaling Technology®, 1:1000), and anti-ETS2 (Santa Cruz Biotechnology, Inc., 1:200). Quantification of protein brand intensities was performed using Image J as described previously34 (link). The band intensities of proteins of interest were first normalized to that of actin (ACTB), yielding relative intensities. The relative intensities of proteins of interest in candidate shRNA-treated cells were further normalized with that in NS shRNA-treated cells.
Guo S., Pridham K.J., Virbasius C.M., He B., Zhang L., Varmark H., Green M.R, & Sheng Z. (2018). A large-scale RNA interference screen identifies genes that regulate autophagy at different stages. Scientific Reports, 8, 2822.
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2

Chromatin Immunoprecipitation for Transcription Factor Binding

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Chromatin immunoprecipitation (ChIP) was performed using the EZ-ChIP kit (17-10086; Millipore). Briefly, cells were cross-linked with 1% formaldehyde for 10 minutes and harvested in SDS lysis buffer, followed by sonication to shear chromatin. Cross-linked protein/DNA was immunoprecipitated with anti-ETS2 (365666X; Santa Cruz Biotechnology), anti-GFP (A-11120; Invitrogen), or control immunoglobulin G. Promoter binding was determined by quantitative PCR.
Luo Y., Jiang N., May H.I., Luo X., Ferdous A., Schiattarella G.G., Chen G., Li Q., Li C., Rothermel B.A., Jiang D., Lavandero S., Gillette T.G, & Hill J.A. (2021). Cooperative Binding of ETS2 and NFAT Links Erk1/2 and Calcineurin Signaling in the Pathogenesis of Cardiac Hypertrophy. Circulation, 144(1), 34-51.
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3

Protein Fractionation and Western Blot Analysis

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Proteins were prepared with radioimmunoprecipitation assay buffer (50 mmol/L Tris-HCl, pH 7.4, 150 mmol/L NaCl, 1 mmol/L EDTA, 0.25% sodium deoxycholate, 0.1% SDS) supplemented with protease and phosphatase inhibitors (A32961; Thermo Fisher). Nuclear and cytoplasmic fractions were isolated using a kit according to the manufacturer’s instructions (78835; Thermo Fisher). A total of 20 to 40 µg of proteins were separated using Criterion gels (Bio-Rad) and transferred to nitrocellulose membranes using the Trans-Blot Turbo Blotting System (Bio-Rad). The membranes were then blocked with 5% nonfat milk or bovine serum albumin, followed by incubation with primary antibodies overnight at 4°C. After 3 washes, the membranes were incubated with secondary antibody and scanned with an Odyssey CLx Imaging system (LI-COR Biosciences). The following antibodies were used: anti-ETS2 (sc-365666; Santa Cruz Biotechnology), anti-phospho-ETS2 (44-1105G; Thermo Fisher), anti-GAPDH (10R-G109a; Fitzgerald), anti-RCAN1.4 (D6694; Sigma), anti-MKP3 (sc-137246; Santa Cruz Biotechnology), anti-lamin A/C (2032; Cell Signaling), anti-NFATc1 (nuclear factor of activated T cells 1; MA3-024; Thermo Fisher), anti-GFP (green fluorescent protein; A-11120; Invitrogen), anti-FLAG (7425; Sigma), anti-Myc (2276; Cell Signaling), anti-Erk1/2 (9107; Cell Signaling), and anti-phospho-Erk1/2 (9101; Cell Signaling).
Luo Y., Jiang N., May H.I., Luo X., Ferdous A., Schiattarella G.G., Chen G., Li Q., Li C., Rothermel B.A., Jiang D., Lavandero S., Gillette T.G, & Hill J.A. (2021). Cooperative Binding of ETS2 and NFAT Links Erk1/2 and Calcineurin Signaling in the Pathogenesis of Cardiac Hypertrophy. Circulation, 144(1), 34-51.
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4

Comprehensive ChIP Assay Antibody Panel

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Antibodies used for ChIP assays were obtained commercially as followed: anti-H3 (ab1791, 1 μg), anti-H3K4me1 (ab8895, 1 μg), and anti-H3K427ac (ab4729, 1 μg) from Abcam; anti-RNA Pol II (N20, sc899, 2 μg), anti-p53 (FL393, sc6243, 2 μg), anti-ETS2 (sc351, 2 μg), anti-p65 (sc372x, 2 μg), and anti-IgG (sc2027, 2 μg) from Santa Cruz Biotechnology. Antibodies used for immunoblotting were obtained as followed: anti-p53 (DO1, sc126, 1:2000 dilution) and anti-β-Actin (sc47778, 1:2000 dilution) from Santa Cruz Biotechnology, and anti-p65 (ab7970, 1:1000 dilution) from Abcam.
Rahnamoun H., Lu H., Duttke S.H., Benner C., Glass C.K, & Lauberth S.M. (2017). Mutant p53 shapes the enhancer landscape of cancer cells in response to chronic immune signaling. Nature Communications, 8, 754.
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5

Western Blot Analysis of ESCC Proteins

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The total proteins of ESCC cells and tissues were extracted with Total Protein Extraction (Sangon Biotech, China). The protein extracts were separated with SDS-PAGE after their concentrations were determined using Bradford protein assay kit (Sangon Biotech, China) and then transferred to polyvinylidine difluoride microporous (PVDF) membranes (Millipore, USA). The blotted membranes were treated with 5% (w/v) skimmed milk in TBST buffer (10 mmol/L Tris, 150 mmol/L NaCl, and 0.2% Tween 20), and incubated for 1 h at room temperature with primary antibody anti Ets2 (Santa Cruz Biotechnology, USA), p-p70S6K (Cell Signaling Technology, USA), Peroxiredoxin 1 (Cell Signaling Technology, USA) and caspase-3 (Abcam, USA), E-cadherin (Abcam, USA) and GAPDH (Cell Signaling Technology, USA). They were then washed and incubated for 1 h with a horseradish peroxidase-linked anti-rabbit secondary antibody (Sangon Biotech, China). Finally, the bands of specific proteins on the membranes were examined by Enhanced Chemiluminescence Kit (Beijing ComWin Biotech Co., Ltd, China) and densitometric analysis was performed with Image J software (NIH).
Li Q., Yang L., Han K., Zhu L., Zhang Y., Ma S., Zhang K., Yang B, & Guan F. (2016). Ets2 knockdown inhibits tumorigenesis in esophageal squamous cell carcinoma in vivo and in vitro. Oncotarget, 7(38), 61458-61468.
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6

Western Blot Analysis of Protein Signaling

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Proteins were prepared with radio-immunoprecipitation assay (RIPA) buffer (50 mM Tris-HCl PH 7.4, 150 mM NaCl, 1mM EDTA, 0.25% sodium deoxycholate, 0.1% SDS) supplemented with protease and phosphatase inhibitors (Thermo Fisher, A32961). Nuclear and cytoplasmic fractions were isolated using a kit according to the manufacturer’s instructions (Thermo Fisher, 78835). 20–40 μg of proteins were separated using Criterion gels (Biorad) and transferred to nitrocellulose membranes using the Trans-Blot® Turbo™ Blotting System (Biorad). The membranes were then blocked with 5% non-fat milk or BSA, followed by incubation with primary antibodies overnight at 4°C. After 3 washes, the membranes were incubated with secondary antibody, and scanned with an Odyssey CLx Imaging system (LI-COR). The following antibodies were used: anti-ETS2 (Santa Cruz Biotechnology, sc-365666), anti-p-ETS2 (Thermo Fisher, 44–1105G), anti-GAPDH (Fitzgerald, 10R-G109a), anti-Rcan1.4 (Sigma, D6694), anti-MKP3 (Santa Cruz Biotechnology, sc-137246), anti-Lamin A/C (Cell Signaling, 2032), anti-NFATc1(Thermo Fisher, MA3–024), anti-GFP (Invitrogen, A-11120), anti-flag (Sigma, 7425), anti-myc (Cell Signaling, 2276), anti-Erk1/2 (Cell Signaling, 9107), anti-p-Erk1/2 (Cell Signaling, 9101).
Luo Y., Jiang N., May H.I., Luo X., Ferdous A., Schiattarella G.G., Chen G., Li Q., Li C., Rothermel B.A., Jiang D., Lavandero S., Gillette T.G, & Hill J.A. (2021). Cooperative Binding of ETS2 and NFAT Links Erk1/2 and Calcineurin Signaling in the Pathogenesis of Cardiac Hypertrophy. Circulation, 144(1), 34-51.
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7

ChIP Assay for ETS2 and GFP

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ChIP was performed using the EZ-ChIP kit (Millipore, 17–10086). Briefly, cells were cross-linked with 1% formaldehyde for 10 minutes and harvested in SDS lysis buffer, followed by sonication to shear chromatin. Cross-linked protein/DNA was immunoprecipitated with anti-ETS2 (Santa Cruz Biotechnology, 365666X), anti-GFP (Invitrogen, A-11120) or control IgG. Promoter binding was determined by quantitative PCR.
Luo Y., Jiang N., May H.I., Luo X., Ferdous A., Schiattarella G.G., Chen G., Li Q., Li C., Rothermel B.A., Jiang D., Lavandero S., Gillette T.G, & Hill J.A. (2021). Cooperative Binding of ETS2 and NFAT Links Erk1/2 and Calcineurin Signaling in the Pathogenesis of Cardiac Hypertrophy. Circulation, 144(1), 34-51.
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