Western blotting was performed as previously described [37 (link)]. In addition, cytoplasmic and nuclear extracts were separated and prepared using NE-PER™ Nuclear and Cytoplasmic Extraction Reagents (Thermo Fisher Scientific, USA) according to the manufacturer’s instructions and a previous study [36 (link)]. Primary antibodies (anti-E-cadherin [Cat# ab15148, 1:500], anti-N-cadherin [Cat# ab18203, 1:1000], anti-ZEB1 [Cat# ab124512, 1:1000], anti-ZEB2 [Cat# ab138222, 1:1000], anti-peroxiredoxin 1/PAG [Cat# ab109498, 1:10,000] and anti-GAPDH [Cat# ab181602, 1:1000] [Abcam, USA]; anti-Vimentin [Cat# D21H3, 1:1000, CST, USA]; anti-H3 [Cat# AH433, 1:1000, Beyotime, China]; and anti-HA Tag [Cat# 26183, 1:10000, Thermo Fisher Scientific]) were used, as well as anti-mouse and anti-rabbit secondary antibodies (IR Dye-labeled secondary antibodies [1:10000, Sigma, USA] and HRP-labeled secondary antibodies [1:10000, CST]). The signals were visualized using an Odyssey Infrared Imaging System (LI-COR Biosciences, USA) or with ECLUltra (New Cell and Molecular Biotech, Suzhou, China).
Anti h3
Manufactured by Beyotime
Sourced in United States
Anti-H3 is a laboratory equipment product designed for research applications. It is used to detect and analyze the presence of H3 histone proteins in biological samples. The core function of Anti-H3 is to provide a reliable and specific method for the identification and quantification of H3 histones, which are important components of chromatin structure and play a crucial role in gene regulation and epigenetic processes.
Automatically generated - may contain errors
Lab products found in correlation
Ne per nuclear and cytoplasmic extraction reagent, by Thermo Fisher Scientific (1 mentions)
Anti e cadherin, by Abcam (1 mentions)
Anti ha tag, by Thermo Fisher Scientific (1 mentions)
Anti n cadherin, by Abcam (1 mentions)
Anti gapdh, by Abcam (1 mentions)
Odyssey infrared imaging system, by LI COR (1 mentions)
Anti zeb2, by Abcam (1 mentions)
Anti zeb1, by Abcam (1 mentions)
Anti phosphoserine, by Merck Group (1 mentions)
Hrp conjugated goat anti mouse igg, by CWBIO (1 mentions)
Peroxidase conjugated goat anti mouse igg, by Merck Group (1 mentions)
Complete protease inhibitor cocktail, by Roche (1 mentions)
Anti myc, by Roche (1 mentions)
Igg affinity chromatography column, by GE Healthcare (1 mentions)
Anti actin, by CWBIO (1 mentions)
2 protocols using anti h3
1
Comprehensive Protein Analysis Protocol
2
Arabidopsis Protein Extraction and Immunoblot Analysis
Check if the same lab product or an alternative is used in the 5 most similar protocols
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Arabidopsis tissues were ground in liquid nitrogen and total proteins were extracted with extraction buffer (50-mM Tris-HCl, pH 7.5, 150-mM NaCl, 10% [v/v] glycerol, 1% [v/v] Nonidet P-40, and 1 × complete protease inhibitor cocktail [Roche]). Cytoplasmic and nuclear fractions were extracted as described previously (Ryu et al., 2007) . Three percent of the cytoplasmic fractions and 10% of the nuclear fractions were used for immunoblot assays. Immunoblotting was performed following standard procedures. The antibodies used were as follows: anti-C3H14 (custom made), anti-Myc (Roche, catalog no. 10653800), anti-phosphoserine (Sigma-Aldrich, catalog no. P3430), anti-MPK4 (Abcam, catalog no. ab49780), anti-ACTIN (Cwbio, catalog no. CW0264M), anti-H3 (Beyotime, catalog no. AF0009), peroxidase-conjugated goat anti-mouse IgG (Sigma-Aldrich, catalog no. A2304), and HRP-conjugated goat anti-mouse IgG (Cwbio, catalog no. C20102S). The anti-C3H14 polyclonal antibody was produced in mice against its full-length protein and purified using an IgG-affinity chromatography column (GE Healthcare) (Institute of Genetics and Developmental Biology, Chinese Academy of Sciences).
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