Brief Protocol Equilibrium dialysis is the most widely accepted method for evaluating plasma protein binding because non-specific binding effects are minimized compared to other methods (such as ultrafiltration). The semipermeable membrane separates the protein-containing compartment from the protein-free com...
Bradford assay protocol For use with AAT Bioquest kit #11118 and #11119 Quick summary Prepare Bradford working solution (90 µL). Add BSA standards or test samples (10 µL). Incubate at room temperature for 5 to 15 minutes. Measure the absorbance at 595 nm and 460 nm; calculate the...
Sensitive—detect down to 5 μg/mL with enhanced protocol No more serial dilutions—kit options include Dilution-Free BSA Protein Standards which eliminate the need to perform serial dilutions when generating a standard curve How the assay works The BCA Protein Assay combines the well-known reduct...
However, its implementation in an assay protocol is complicated by pipetting and dilution steps, evaluation of replicates, blank-corrections and other factors. These steps frequently cause confusion with regard to the calculations that are...
SRM have been used for the analysis of proteins and peptides in body fluids such as blood plasma/serum[154–157]or urine[158,159]. It has been successfully applied to the analysis of Hb variants relevant in hemoglobinopathies[154,155]. Moat et al. developed a protocol for new-born blood...
Scan the array and extract the data as mentioned in the previous small-molecule CSI protocol section (steps 5 and 6). Show moreView chapter Book series 2011, Methods in EnzymologyJoshua R. Tietjen, ... Aseem Z. Ansari Chapter Lactose Repressor Protein: Functional Properties and Structure II ...
Minimum detection level of 5 µg/ml with the enhanced protocol Adaptable to microplates Less protein-to-protein variation than dye-binding methods BCA Protein Assay Applications: Studying protein:protein interactions Measuring column fractions after affinity chromatography Estimating percent recovery...
Residual Protein A Protocol This protocol is intended as a guide only, for full experimental details please read the reference provided. Introduction Duringbioprocessing, most therapeuticmonoclonal antibodies (mAbs)are isolated from cell culture media using Protein A chromatography.1During purification, ...
In vitro binding assay In vitro binding assays were performed following the RNA Bind-n-Seq protocol72with the following modifications. Recombinant Cp was immobilized on Strep-TactinXT coated magnetic beads (20 pmole protein/μl bead slurry) in buffer w (100 mM Tris-Cl pH 8.0, 150 mM ...
Oligonucleotides for affinity purification were ordered as custom-synthesized oligos from Integrated DNA Technologies (IDT; Supplementary Table1). DNA and nucleosome affinity purifications were performed using a filter plate-based workflow described first by Hubner et al18. The essential protocol is descri...