Monoclonal (mAb) and polyclonal (pAb) antibodies were used to detect the following cellular proteins: rabbit mAb-lamin A/C (EPR4100, Abcam), mouse mAb-IE1p72 (P63-27), mouse mAb-pp28, and mouse mAb-UL53.01 (kindly provided by Stipan Jonjic and Tihana Lenac Rovis, University of Rijeka, Croatia). Alexa Fluor 555- and 647-conjugated antibodies were used as secondary antibodies for indirect immunofluorescence staining (Molecular Probes, Eugene, OR, USA).
Epr4100
Manufactured by Abcam
Sourced in United Kingdom
The EPR4100 is an electron paramagnetic resonance (EPR) spectrometer designed for research applications. It provides a stable and reliable platform for the detection and characterization of paramagnetic species, such as free radicals, transition metal ions, and other unpaired electron systems. The EPR4100 offers high-sensitivity measurements and versatile operating modes to support a wide range of scientific investigations.
Automatically generated - may contain errors
Lab products found in correlation
Bca protein assay kit, by Thermo Fisher Scientific (2 mentions)
Normal goat serum (ngs), by Jackson ImmunoResearch (1 mentions)
Axiovision windows software version 4, by Zeiss (1 mentions)
Ne per nuclear and cytoplasmic extraction kit, by Thermo Fisher Scientific (1 mentions)
A3682, by Merck Group (1 mentions)
A9169, by Merck Group (1 mentions)
Ecl reagent, by Bio-Rad (1 mentions)
Iblot2, by Thermo Fisher Scientific (1 mentions)
Nupage 4 12 bis tris mini protein gel, by Thermo Fisher Scientific (1 mentions)
Anti gapdh, by Merck Group (1 mentions)
Protease inhibitor cocktail, by Roche (1 mentions)
4 protocols using epr4100
1
Antibody Detection of Cellular Proteins
2
Tracking Administered SHED in Kidney
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To evaluate the tracking of administered SHED, kidney samples were collected on days 2, 7, and 14. Paraffin sections (1μm thick) were incubated with rabbit anti-human Lamin A/C antibody (EPR4100; Abcam plc, Cambridge, UK) subsequent to blocking with 10% normal goat serum (Jackson Immuno-Research Laboratories, Inc., USA) for 30 min. The primary antibody was visualized using the Histofine Simple Stain PO(R) kit (Nichirei, Tokyo, Japan) according to the manufacturer’s instructions. Slides were counterstained with hematoxylin [23 (link)]. Tissue images were taken with a Zeiss Z1 microscope and Axiovision Windows software version 4.4 (Carl Zeiss, Oberkochen, Germany).
3
AhR Translocation and Target Engagement Assay
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Hep G2 cells were treated with 100 nM TCDD in the absence (negative control) or presence of BAY 2416964 for 4 hours. Cytoplasmic and nuclear protein fractions were extracted using the NE-PER Nuclear and Cytoplasmic Extraction Kit (Thermo Fisher Scientific). Agonist-induced translocation of AhR from the cytoplasm to the nucleus was analyzed by western blotting using primary antibodies selective against AhR (Clone EPR7119(N)(2)), alpha tubulin (DM1A), and lamin A/C (EPR4100) (all from Abcam) and quantified via densitometric analysis using the ImageJ software. Target engagement was determined by cellular thermal shift assay (CETSA) using MDA-MB-231 cells treated with increasing concentrations of BAY 2416964.
4
Immunoblotting Protocol for Protein Analysis
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For immunoblotting, RIPA buffer (150 mM NaCl, 50 mM Tris HCl, pH 8, 0.1% SDS, 1% Nonidet P-40, 0.5% sodium deoxycholate, and protease inhibitor cocktail [Roche]) was added to cell fractions or whole cells in lysis buffer for 30 min on ice and pelleted. Protein concentration was determined using a BCA protein assay kit (Thermo Fisher Scientific). Proteins were separated in NuPAGE 4%–12% Bis-Tris mini protein gels (Thermo Fisher Scientific) and electro-transferred onto PVDF membranes using iBlot 2 (Thermo Fisher Scientific). Membranes were blocked with 5% milk in TBS with Tween-20 for 2 h and incubated overnight at 4°C with primary antibodies—anti-ELAVL1 (3A2, sc-5261, Santa Cruz Biotechnology), anti-GAPDH (G8795, Sigma-Aldrich), anti-Lamin A + Lamin C (EPR4100, Abcam), anti-PATZ1 (sc390577, Santa Cruz Biotechnology), and anti-MAP3K5 (SAB4300398, Sigma-Aldrich). Membranes were washed and incubated with horseradish peroxidase-conjugated goat antimouse or antirabbit antibodies (A3682 and A9169, Sigma-Aldrich). After washing, proteins were detected with ECL reagent (Bio-Rad), according to the manufacturer's protocol.
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