Table 2 Merits and demerits of the techniques of western blotting
From: Emerging techniques of western blotting for purification and analysis of protein
Methods | Merits | Demerits | References |
|---|---|---|---|
Capillary and microchip electrophoresis (MCE) | Increased sensitivity and better precision | Time-consuming and usually identifying only one protein at a time | |
DigiWest | Enhanced sample quality, lower lysate requirements/target monitoring, and low antibody quantity required | These include the alteration of target proteins by biotinylation, the need for advanced reagents and facilities, the initial phase costs, issues about conversion of digital DigiWest test to Western blot mimics | [14] |
Micro-loader | Simple and efficient It requires very small numbers of filling samples for the identification of signals | Needs a micro-loader device that installs samples into a funnel-like framework | [1] |
Thin-film direct coating with suction (TDCS) | An exceptionally sensitive and fast control system for quantitative studies of multiple antigen–antibody interactions needs a shorter running time | Still in installation would only be a few years before any laboratory uses it | [15] |
Polyacrylamide gel improvement | Better shelf life than pre-manufactured gels | - | [38] |
Retention improvement of hormones of peptide on the membrane of blotting | By use of a citrate buffer enhanced the insulin signal significantly | – | [38] |
Blotting of diffusion | They're a quick and easy step to generate multiple blots by diffusion. Multiple proteins can be analyzed on specific blots formed by the specific gel | – | [39] |
Analysis of western blot utilizing molecular weight marker detecting secondary antibody | Provide fast and effective testing of protein size | The application of this class of markers is limited to fact that these can be used for accurate molecular mass measurement in non-denaturable conditions | |
Total and target protein co-detection by immunoblotting of fluorescent ECL and labelling of Cydye | At the same time, this process allows the immune identification of unique target proteins as well as the optimal protein expression | While modification of CyDye proteins will have an effect on antibody-antigen interconnections, the mechanism is highly sensitive | |
Micro western | Further used diagnose the problem of gp120 protein purification of blood plasma | Although not yet commonly available, the Micro-Western is likely a suitable choice to exploit as a diagnostic tool for cancer biology and infectious diseases | [48] |
System of iBind western | Using a comparatively inexpensive semi-automated western blot analysis system that allocates reagents and antibodies to different compartments using sequential flow technology | Major concerns this method include poor performance of computer and the need for advanced materials including a Bind Card | [49] |
Blot cycler | Are reduced prerequisites for antibodies than standard blots, the primary antibody is stored and the utilization increases instantly | [50] | |
System of SNAP id | It takes around 30 min for washing, incubation of antibody, and blocking | However, the signal-to-noise ratio of bands observed was not as high as the traditional Western blot approach | [51] |