Ptm 301

Manufactured by PTM Biolabs

Sourced in China

The PTM-301 is a high-performance laboratory centrifuge designed for a variety of applications. It features a compact and durable construction, and can accommodate a range of rotor sizes and sample volumes.

Automatically generated - may contain errors

Lab products found in correlation

Ptm 501, by PTM Biolabs (4 mentions) Anti crotonyllysine, by PTM Biolabs (2 mentions) Odyssey system, by LI COR (1 mentions) Ecl substrate, by Thermo Fisher Scientific (1 mentions) Goat anti rabbit igg hrp, by Santa Cruz Biotechnology (1 mentions) Alexa 488, by Thermo Fisher Scientific (1 mentions) Dapi dye, by Vector Laboratories (1 mentions) Anti h3 ab1791, by Abcam (1 mentions) Epiquik total histone extraction kit, by Epigentek (1 mentions) Ptm 901, by PTM Biolabs (1 mentions) Ptm 1151, by PTM Biolabs (1 mentions) Ptm 401, by PTM Biolabs (1 mentions) Anti h3k9ac, by Thermo Fisher Scientific (1 mentions) Ab1791, by Abcam (1 mentions) Ptm 419, by PTM Biolabs (1 mentions) Ptm 105, by PTM Biolabs (1 mentions) Hiseq 2500, by Illumina (1 mentions)

6 protocols using ptm 301

Histone H4 Acetylation Assay

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1 μg of human recombinant histone H4 (New England BioLabs, Cat# M2504S) was incubated with 50 μM n-butyryl- or isobutyryl-CoA and 0.2 μM of HAT1 at 30°C for 1 h. The reaction mixture was boiled in SDS-PAGE gel loading buffer and resolved on a 15% polyacrylamide gel followed by wet membrane transfer to a nitrocellulose (NC) membrane. The NC membrane was blocked with 5% non-fat milk in Tris-buffered saline + 0.1% Tween-20 (TBST) for 1 h at room temperature. Anti-butyryllysine antibody (PTM BioLabs, PTM#301) at 1:2000 dilution was incubated with the membrane overnight at 4°C. The membrane was washed with TBST buffer for three times and incubated with the goat anti-rabbit IgG-HRP (Santa Cruz Biotechnology, Cat# sc-2004) with 1:3000 dilution at room temperature for 1 h. The membrane was then washed and subjected to chemiluminescence detection with the ECL substrate (ThermoFisher, Cat# 32209) on a LI-COR Odyssey system (LI-COR Biosciences).

Zhu Z., Han Z., Halabelian L., Yang X., Ding J., Zhang N., Ngo L., Song J., Zeng H., He M., Zhao Y., Arrowsmith C.H., Luo M., Bartlett M.G, & Zheng Y.G. (2020). Identification of lysine isobutyrylation as a new histone modification mark. Nucleic Acids Research, 49(1), 177-189.

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Rice Histone Protein Analysis

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Western blotting was performed as previously described (Liu et al. 2018 (link)). Immunofluorescence analysis was performed using the method described by Gong et al. (Gong et al. 2009 (link)). The rice histone proteins were separated electophoretically in denaturing gels by SDS-PAGE (5%/12%). The antibodies used in this study were rabbit pan anti-Kbu antibody (1:5000; PTM BioLabs, HangZhou China, PTM-301) and rabbit anti-H3 antibody (1:10,000; PTM BioLabs, HangZhou China, PTM-1001); the goat secondary anti-rabbit antibody is conjugated with Alexa 488 (Invitrogen, A11008). Chromosomes were counterstained with DAPI dye (Vector Laboratories, H-1200).

Liu S., Liu G., Cheng P., Xue C., Zhou Y., Chen X., Ye L., Qiao Z., Zhang T, & Gong Z. (2019). Genome-wide Profiling of Histone Lysine Butyrylation Reveals its Role in the Positive Regulation of Gene Transcription in Rice. Rice, 12, 86.

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Histone Extraction and Quantification in Plants

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Histone-enriched fractions were extracted from seedlings of Arabidopsis, rice, maize, and tobacco plants using EpiQuik Total Histone Extraction Kit (Epigentek USA, OP-0006-100). The histone-enriched fractions were used for immunoblot analysis; antibodies used in this study were anti-H3 (ab1791; Abcam), anti-H3K9ace (07-352; Millipore), anti-butyryllysine (PTM-301; PTM Biolabs), and anti-crotonyllysine (PTM-501; PTM Biolabs). Immunoblotting results were quantified using ImageJ (v1.6.0_24).

Lu Y., Xu Q., Liu Y., Yu Y., Cheng Z.Y., Zhao Y, & Zhou D.X. (2018). Dynamics and functional interplay of histone lysine butyrylation, crotonylation, and acetylation in rice under starvation and submergence. Genome Biology, 19, 144.

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Histone Post-Translational Modification Profiling

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2 μg of full-length acylated histones H3 or H4 from in vitro reactions were spotted onto a nitrocellulose membrane. The membrane was blocked with either 5% BSA or 5% nonfat milk and incubated with the primary antibodies at 1:1,000 dilution (pan anti-crotonyl-lysine: PTM-501, pan anti-propionyl-lysine: PTM-201, pan anti-malonyl-lysine: PTM-901, pan anti-succinyl-lysine: PTM-401, pan anti-butyryl-lysine: PTM-301, pan anti-glytaryl-lysine: PTM-1151, all purchased from PTM-Biolabs, Hangzhou, China) according to the manufacturers’ instructions overnight at 4 °C. The membrane was washed with TBS-T three times for 10 min each, incubated with secondary antibody (Goat anti-Rabbit IgG Fc, Pierce 31463) at a 1:10,000 dilution for 60 min at room temperature and then probed with ECL Western Blot Substrate (Pierce).

Simithy J., Sidoli S., Yuan Z.F., Coradin M., Bhanu N.V., Marchione D.M., Klein B.J., Bazilevsky G.A., McCullough C.E., Magin R.S., Kutateladze T.G., Snyder N.W., Marmorstein R, & Garcia B.A. (2017). Characterization of histone acylations links chromatin modifications with metabolism. Nature Communications, 8, 1141.

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Epigenetic Chromatin Modifications Analysis

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Two grams of rice seedling leaves was cross-linked by 1% formaldehyde and used for chromatin extraction. After sonication, chromatin fragments were incubated with antibody (anti-H3K9ac, anti-Kbu, and anti-Kcr)-coated beads (Invitrogen/Life Technologies; 10001D) overnight. Specificity of anti-Kbu and anti-Kcr was tested by dot blots (Additional file 1: Figure S8). After extensive washing, immunoprecipitated chromatin was de-cross-linked and retrieved for qPCR or sequencing. anti-H3K9ace (07-352; Millipore), anti-butyryllysine (PTM-301; PTM Biolabs), and anti-crotonyllysine (PTM-501; PTM Biolabs) antibodies were used.

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ChIP-Seq Profiling of Histone Modifications

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Epi-ID was performed as described^{23 (link), 25 (link)}, except that for ChIP the following antibodies were used: anti-pan-K-acetylation (PTM-105; PTM-Biolabs), anti-pan-K-crotonylation (PTM-501;PTM Biolabs), anti-pan-K-butyrylation (PTM-301; PTM Biolabs), anti-pan-K-succinylation (PTM-419; PTM Biolabs) and anti-H3 (ab1791; Abcam). Deep-sequencing was performed on a single-end flowcell Illumina Hi-Seq2500.

Kollenstart L., van der Horst S.C., Vreeken K., Janssen G.M., Martino F., Vlaming H., van Veelen P.A., van Leeuwen F, & van Attikum H. (2021). Epigenetics Identifier screens reveal regulators of chromatin acylation and limited specificity of acylation antibodies. Scientific Reports, 11, 12795.

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