ProteinSimple Jess (biotechne) was used for low-input digital capillary Western blot-like protein analysis. FACS-purified iNs were plated on Geltrex-coated wells and harvested with PBS. Pellets were then resuspended in RIPA Lysis and Extraction Buffer (Thermo Fisher) containing the cOmplete EDTA-free Protease Inhibitor Cocktail and PhosSTOP phosphatase inhibitors (Sigma Aldrich). Samples were pooled from 9 CTL and AD lines and analyzed using the 12–230 kDa Jess Separation Module. Concentrations of primary antibodies (CDKN2A 1:100) were adjusted to the technology. All assays were run using the standard default settings provided by ProteinSimple Inc., with the exception of an increased 60-min incubation time for the primary antibodies. Data analysis was performed using Compass software (biotechne).
Proteinsimple jess
Manufactured by Bio-Techne
Sourced in United States
The ProteinSimple Jess is a lab equipment product that enables automated protein analysis. It is designed to provide consistent and reliable results for researchers in various fields.
Automatically generated - may contain errors
Lab products found in correlation
Ripa lysis and extraction buffer, by Thermo Fisher Scientific (3 mentions)
Phosstop phosphatase inhibitor, by Merck Group (2 mentions)
Complete edta free protease inhibitor cocktail, by Merck Group (1 mentions)
Edta free protease inhibitor cocktail, by Merck Group (1 mentions)
Phosstop, by Merck Group (1 mentions)
Anti pdh antibody, by Santa Cruz Biotechnology (1 mentions)
Cytation 5, by Agilent Technologies (1 mentions)
Halt protease and phosphatase inhibitor, by Thermo Fisher Scientific (1 mentions)
4 protocols using proteinsimple jess
1
Digital Capillary Western Blot for Low-Input Neuronal Samples
2
Quantifying PDH Protein in iNs
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PDH protein was quantified in lysed pellets of FACS-purified iNs using the ProteinSimple Jess (Biotechne) and the 12-230 kDa Jess Separation Module in the NIR channel. Pellets were lysed in RIPA Lysis and extraction Buffer (ThermoFisher) containing cOmplete EDTA-free Protease Inhibitor Cocktail (Merck, 11873580001) and PhosSTOP (Merck, 4906845001). Anti-PDH antibody (Santa Cruz, sc-377092, 1:50) was incubated for 60 minutes, followed by standard default run settings provided by ProteinSimple. Data analysis was performed using Compass software.
3
Quantifying Tau Proteomic Profiles in iNeurons
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ProteinSimple Jess (biotechne) was used for low-input digital capillary western blot-like protein analysis. FACS-purified iNs were plated on Geltrex-coated wells and harvested with PBS. Pellets were then resuspended in RIPA Lysis and Extraction Buffer (Thermo Fisher) containing the cOmplete EDTA-free Protease Inhibitor Cocktail and PhosSTOP phosphatase inhibitors (Sigma Aldrich). Samples for tau analysis were pooled from 3 control and AD lines and analyzed using the 12-230 kDa Jess Separation Module. Concentrations of primary antibodies (ACLY 1:50, beta-tubulin 1: 200, PKM2 1:50, P-PKM2 1:100) were adjusted to the technology. Different antibody dilutions (indicated in the figure) were used for TG5, PHF1, Alz50 (kindly provided by Dr. Peter Davis), and AT100. All assays were run using the standard default settings provided by ProteinSimple Inc., with the exception of an increased 60-min incubation time for the primary antibodies. Data analysis was performed using Compass software (biotechne).
4
Protein Quantification and Analysis
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Cells and retina proteins were collected in RIPA buffer with 1% HALT protease and phosphatase inhibitors (Thermo Fisher Scientific, Waltham, MA, USA). The total protein concentration was measured using the Bicinchoninic Acid Assay Kit (Pierce, ThermoFisher Scientific) and read on a Cytation 5 (Biotek, Winooski, VT, USA) plate reader. Proteins were analyzed by capillary tube-based electrophoresis immunoassay using the ProteinSimple Jess instrument (Bio-Techne, Minneapolis, MN, USA) and each protein was normalized to total protein in the sample capillary. Antibodies used for protein analysis are listed in Table 1 .
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