Treated cells were harvested and washed with ice-cold phosphate-buffered saline. Cell lysates were prepared in RIPA buffer with 1 X ProteoBlock protease inhibitor cocktail (Fermentas, Glen Burnie, MD, USA) and 1 mmol·L−1 PMSF. For separation of nuclear and cytoplasmic fractions for extraction, the NE-PER nuclear and cytoplasmic extraction kit (Thermo Fisher Scientific Inc., Rockford, IL, USA) was utilized. Protein samples (20 μg) were separated by SDS-PAGE. Primary anti-hnRNP C1/C2 (sc-32308; Santa Cruz Biotechnology, Santa Cruz, CA, USA) and secondary antibodies were used at 1∶1 000. I-Block™ blocking reagent (Applied Biosystems, Foster City, CA, USA) was used during incubation steps.
I block blocking reagent
Manufactured by Thermo Fisher Scientific
Sourced in United States
The I-Block™ blocking reagent is a protein-based solution designed to minimize non-specific binding in immunoassays and other research applications. It functions by occupying non-specific binding sites on surfaces, thereby reducing the background signal and improving the signal-to-noise ratio.
Automatically generated - may contain errors
2 protocols using i block blocking reagent
1
Western Blot Analysis of hnRNP C1/C2
2
Extraction and Separation of Cell Fractions
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