Rabbit polyclonal antibodies against H2B (abcam, ab1790, 1:2000 dilution for WB), H3K9ac (Merck, 07-352, 1:1000 dilution for WB), H3K18ac (abcam, ab1181, 1:1000 dilution for WB), and pan-Kma (PTM-Biolabs, PTM-901, 1:1000 dilution for WB), a rabbit monoclonal antibody against p300 (Cell Signaling Technology, 54062, 1:1000 dilution for WB), and mouse monoclonal antibodies against H2BK120ac (1 μg/mL for WB) and H3K27ac (Merck, 05-1334, 1:1000 dilution for WB) were used for western blot and ChIP assay. The monoclonal antibody against H2BK120ac was generated as described previously15 (link). Secondary antibodies, anti-rabbit HRP (Cell Signaling Technology, 7074S, 1:5000 for WB) and anti-mouse HRP (GE Healthcare Life Sciences, NA931V, 1:10000 for WB or Jackson ImmunoResearch, 515-035-003, 1:10000 for WB) were also used for western blot. Normal rabbit IgG (SantaCruz, sc-2027) and Normal mouse IgG (SantaCruz, sc-2025) were also used for ChIP assay. The amounts of antibodies used for ChIP assay were indicated in Supplementary Methods section .
Ptm 901
Manufactured by PTM Biolabs
Sourced in China
The PTM-901 is a high-performance laboratory centrifuge designed for a variety of applications. It features a brushless motor and digital display for precise speed and time control. The PTM-901 can achieve speeds up to 14,000 RPM, making it suitable for a range of sample separation and preparation tasks.
Automatically generated - may contain errors
Lab products found in correlation
Na931v, by Jackson ImmunoResearch (1 mentions)
Normal mouse igg, by Santa Cruz Biotechnology (1 mentions)
Sc 2027, by Santa Cruz Biotechnology (1 mentions)
Normal rabbit igg, by Santa Cruz Biotechnology (1 mentions)
Horseradish peroxidase conjugated goat anti rabbit igg, by Thermo Fisher Scientific (1 mentions)
Lotus tetragonolobus lectin, by Vector Laboratories (1 mentions)
H 3300, by Vector Laboratories (1 mentions)
Alexa fluor 647 anti rabbit igg, by Thermo Fisher Scientific (1 mentions)
Paraformaldehyde (pfa), by Electron Microscopy Sciences (1 mentions)
Ptm 1151, by PTM Biolabs (1 mentions)
Ptm 501, by PTM Biolabs (1 mentions)
Ptm 301, by PTM Biolabs (1 mentions)
Ptm 401, by PTM Biolabs (1 mentions)
Ab93678, by Abcam (1 mentions)
Alexa fluor 488 goat anti rabbit igg green, by Thermo Fisher Scientific (1 mentions)
P36931, by Thermo Fisher Scientific (1 mentions)
Dp 70 digital microscope, by Olympus (1 mentions)
H1009, by Merck Group (1 mentions)
5 protocols using ptm 901
1
Antibodies for Histone Modifications Analysis
2
Detection of Malonylated Proteins
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Proteins (20 μg) of tachyzoites were electrophoresed on 12% SDS-PAGE and transferred to PVDF membrane. TBST buffer (25 mM Tris–HCl, 150 mM NaCl, 0.1% Tween20, pH 8.0) with 5% BSA was used to block the membrane for 60 min. Then, the membrane was incubated with anti-malonyllysine antibody (1:500, catalog no. PTM-901; PTM Biolabs, Hangzhou, China) in TBST with 2.5% BSA overnight at 4°C. The membrane was washed three times with TBST, followed by incubation with horseradish-peroxidase-conjugated Goat anti-Rabbit IgG (1:5000; Thermo) for 60 min at room temperature. After washing the membrane three times, an ECL kit was used for protein visualization.
3
Kidney Malonylation Analysis via Immunofluorescence
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Kidney sections were fixed in 4% paraformaldehyde (Electron Microscopy Sciences) overnight at room temperature. Sections were paraffin embedded and cut into 3-μm sections for analysis. Slides were deparaffinized and incubated at 95 °C in antigen-unmasking solution (H-3300, Vector laboratories) for 2 h. Sections were blocked and permeabilized with 10% donkey serum and 0.5% Tween-20, respectively. Sections were incubated with pan anti-malonyllysine (PTM-901, PTM-biolabs) and Lotus Tetragonolobus Lectin tagged with Fluorescein (FL-1321-2, Vector laboratories) overnight at 4 °C. Sections were then incubated with anti-rabbit IgG Alexa Fluor 647 (A-31573, Thermo Fisher). Resulting sections were visualized with Leica SP5 Confocal TCS Microscope. Semiquantitation of immunofluorescence malonylation signals was analyzed with Leica Application Suite Lite 2.6.3.
4
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).
5
Immunohistochemistry of Kidney Tissue
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Formalin-fixed, paraffin-embedded 2- to 3-micron sections were obtained from renal biopsies in the archives of the University of Michigan Department of Pathology. Human kidney biopsies were dewaxed, rehydrated, and subjected to heat-induced antigen retrieval by incubating in 0.1 M citrate buffer (pH 6.0, ab93678, Abcam) using the 2100 Antigen Retriever. Endogenous peroxidase was blocked with 0.3% hydrogen peroxide (H1009, Sigma) in Tris-buffered saline (TBS, pH 7.4) for 30 min. Sections were blocked with 10% normal goat serum in 5% bovine serum albumin at room temperature for 2 h. Sections were incubated with primary antibodies, Kmal (1:150, PTM-901, PTM Biolabs) or Sirt5 (1:250, PA5-31029, Invitrogen), diluted in Da Vinci Green (PD900L, Biocare Medical) at 4 ˚C overnight. Antibody bindings were detected by using secondary antibody, Alexa Fluor 488 goat anti-rabbit IgG (green, 1:300, A11034) at room temperature for 1 h. The sections were mounted with gold antifade reagent with DAPI (P36931, Invitrogen). An Olympus DP70 Digital Microscope was used to take pictures by ×20 magnification lenses. Fluorescent signal was quantified with ImageJ.
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