Purified proteins of mbE1E2, sE1E2.LZ, and sE2 were separated by a precast, 4-20% Mini-PROTEAN TGX stain-free gels on a Mini-PROTEAN Tetra cell electrophoresis instrument (Bio-Rad). In reducing conditions, each sample was incubated with loading dye (4× Laemmli buffer + 10% β-mercaptoethanol) (Bio-Rad) and heated to 95 °C. In nonreducing conditions, each sample was incubated with Laemmli buffer and heated to 37 °C. For Western blot detection, the purified protein samples on SDS/PAGE were transferred onto Trans-Blot Turbo Mini nitrocellulose membranes (Bio-Rad). The membranes were then probed using the anti-HCV E2 mAb HCV1 at 5 μg/mL and anti-HCV E1 mAb H-111 at 10 μg/mL followed by detection using a secondary goat anti-human IgG-HRP conjugate (Invitrogen) at a 1:5,000 dilution and the Western ECL substrate (Bio-Rad). All gels were imaged using the ChemiDoc system (Bio-Rad).
Trans blot turbo mini nitrocellulose membranes
Manufactured by Bio-Rad
Sourced in United States
The Trans-Blot Turbo Mini nitrocellulose membranes are a type of laboratory equipment used for protein transfer and blotting applications. They provide a surface for the immobilization of proteins, enabling the detection and analysis of specific proteins within a sample.
Automatically generated - may contain errors
Lab products found in correlation
Pierce western blotting substrate plus, by Thermo Fisher Scientific (2 mentions)
Mini protean tgx gel, by Bio-Rad (2 mentions)
Anti rabbit igg hrp linked antibody, by Cell Signaling Technology (2 mentions)
Exoquick, by System Biosciences (2 mentions)
Loading dye, by Bio-Rad (1 mentions)
Goat anti human igg hrp conjugate, by Thermo Fisher Scientific (1 mentions)
Western ecl substrate, by Bio-Rad (1 mentions)
Chemidoc system, by Bio-Rad (1 mentions)
Image studio lite software, by LI COR (1 mentions)
Odyssey infrared imaging system, by LI COR (1 mentions)
Trans blot turbo transfer system, by Bio-Rad (1 mentions)
Anti cd63, by Santa Cruz Biotechnology (1 mentions)
Anti c met, by Cell Signaling Technology (1 mentions)
Anti macc1, by Santa Cruz Biotechnology (1 mentions)
Anti halotag, by Promega (1 mentions)
Anti β actin, by Cell Signaling Technology (1 mentions)
4 protocols using trans blot turbo mini nitrocellulose membranes
1
HCV Envelope Protein Analysis
2
Western Blot Analysis of Pancreatic Tumor
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Tumor lysates were made using normal human pancreas tissue and the pancreatic cancer cell line BxPC-3-GFP xenograft. Western blotting was performed as previously described.18 (link) Protein samples were isolated and transferred to Trans-Blot Turbo mini nitrocellulose membranes (Bio-Rad, Hercules, CA, Cat. 1704270) using the Bio-Rad Trans-Blot Turbo transfer system. The membrane was then incubated with the IRDye700DX-conjugated antibody at 4 °C overnight. The membrane was scanned with an Odyssey infrared imaging system (model 9120; LI-COR), and detection and quantification of band intensities were conducted using Image Studio Lite software (version 5.2; LI-COR). β-Actin was used as standard.
3
Exosomal Protein Characterization by Western Blot
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The pelleted exosomes extracted using ExoQuick (System Biosciences) were resuspended in phosphate-buffered saline following the protein analysis. The proteins were loaded on Mini-PROTEAN® TGX™ gel (Bio-Rad Laboratories, Hercules, CA, USA) and electrophoretically transferred onto Trans-Blot® Turbo™ Mini Nitrocellulose membranes (Bio-Rad Laboratories). The membranes were blocked and incubated overnight at 4°C with the primary antibody anti-CD63 (1:200; Santa Cruz Biotechnology, Fremont, CA, USA). Proteins were visualized with horseradish peroxidase-conjugated secondary antibodies (anti-rabbit IgG, HRP-linked antibody; Cell Signaling Technology, Tokyo, Japan) followed by chemiluminescence detection (Pierce Western Blotting Substrate Plus; Thermo Scientific, Tokyo Japan).
4
Western Blot Analysis of MACC1, c-Met, and β-Actin
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Extraction Buffer (GE Healthcare Ltd., Buckinghamshire, Uk). The protein concentration was determined using a Pierce Protein 660 nm Protein Assay (Thermo Fisher Scientific k.k., kanagawa, Japan). Equal amounts of protein were separated on Mini-PROTEAN ® TGX™ gel and electrophoretically transferred onto Trans-Blot ® Turbo™ Mini Nitrocellulose membranes (both from Bio-Rad Laboratories, Inc., Hercules, CA, USA). The membranes were blocked and incubated overnight at 4˚C with primary antibody; anti-MACC1 (1:200; Santa Cruz Biotechnology, Inc., Santa Cruz, CA, USA), anti-c-Met (1:1,000), anti-β-actin (1:500) (both from Cell Signaling Technology Japan k.k.) or anti-HaloTag ® (1:1,000; Promega k.k.). Proteins were visualized with horseradish peroxidase-conjugated secondary antibodies (anti-rabbit IgG, HRP-linked antibody, Cell Signaling Technology Japan k.k.; anti-goat IgG-HRP, Santa Cruz Biotechnology, Inc.) followed by chemiluminescence detection (Pierce Western Blotting Substrate Plus; Thermo Fisher Scientific k.k.).
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