Functional Molecular Lumino-Materials to Probe Serum Albumins: Solid Phase Selective Staining Through Noncovalent Fluorescent Labeling

Abstract

Selective staining of human serum albumin protein in gel electrophoresis over wide range of other protein(s) is extremely important because it contains more than 60% volume of serum fluid in human body. Given the nonexistence of suitable dye materials for selective staining of serum albumins in gel electrophoresis, we report a new class of easy synthesizable and low molecular weight staining agents based on 3-amino-N-alkyl-carbazole scaffold for selective staining of serum albumins in solid phase. A detailed structure-efficiency relationship (SER) study enabled us to develop two such potent functional molecular probes which stain both human and bovine serum albumin selectively in gel electrophoresis in the presence of other proteins and enzymes. The present gel staining process was found to be very simple and less time-consuming as compared to the conventional coomassie blue staining which in turn makes these probes a new class of serum albumin-specific staining materials in proteome research. Moreover, these molecular lumino-materials can detect serum albumins at subnanomolar level in the presence of broad spectrum of other proteins/enzymes in aqueous buffer (99.9% water, pH = 7.3) keeping the protein secondary structure intact. Our experimental and the docking simulation results show that these probes bind preferentially at 'binding site I' of both the serum proteins.

Host–Host Interactions Control Self‐assembly and Switching of Triple and Double Decker Stacks of Tricarbazole Macrocycles Co‐assembled with anti‐Electrostatic Bisulfate Dimers

“…The crude solid mixture was recrystallized from hexanes to give an orange solid product (1.64 g, 5.55 mmol, 94 % yield). The 1 H NMR spectrum was identical to previous reports …”

“…The 1 HNMR spectrum was identical to previous reports. [34] 9-Nonyl-3-nitrocarbazole (C 9 -3):N itro C 9 -3 was prepared as an orange solid following the same procedure as C 6 -3 in 92 %y ield from 143.48, 141.57, 140.50, 127.30, 122.79, 122.49, 121.56, 120.94, 120.67, 117.30, 109.64, 108.20, 43.58, 31.74, 29.37, 29.28, 29.14, 28.86, 27.19, 22.57, 14.03. HRMS (EI): 339.1994 [M+ +H] + ;f ound:339.2070.…”

Hydrophobicity-induced prestaining for protein detection in polyacrylamide gel electrophoresis

“…These phenomena might be attributed to the differences in the microenvironment between a-La and BSA/LDH. 17 Finally, a controlled gel experiment was carried out by poststaining with CBB, which had a higher detection limit with the background signal (Fig. 5B), clearly demonstrating the feasibility of the developed fluorescence prestaining method for the detection of proteins.…”

p‐TsOH‐mediated, Versatile, and Efficient Approach for the Synthesis of Triazolyl‐Carbazoles from Nitrovinylcarbazoles and Azide via 1, 3‐Dipolar Cycloaddition

“…As shown in Scheme , the commercially available carbazole ( 1 ) was treated with different alkylating agents such as methyl iodide, ethyl bromide, and butyl bromide in acetone at RT using K 2 CO 3 as a base to give the corresponding N ‐alkylated carbazoles ( 2 ) . This on Vilsmeier–Haack formylation at RT for 1 h resulted in the formation of carbazole‐3‐carbaldehyde ( 3 ) .…”