I am interested to know how it compares to standard SDS-PAGE/Western Blotting procedures in terms of sensitivity (using grey x-ray films), separation quality, handling and overall costs. Any critical issues?
Hi Ehlert,
Both experiments are sensitive, but W.B is much more sensitive and specific. With the W.B you can detect your protein molecular weight and expression level, unlike SDS unless you know where it is. In term of separation, they are good enough if they are run in a proper waye. They are not expensive specially if you prepare the gel and some other reagents in your lab. HOPE, that would help. Good luck
From my experiences, I have not gotten a big issue with these experiments.
The cost is similar, though you are made more aware of it with the Simple Wes because of the upfront cost of the kit. You can dilute antibodies quite a bit more with the Wes than SDS-PAGE, so you save a good deal of money there. One disadvantage of the Wes is that samples are run in individual capillaries, thus there can be slight variability between capillaries-- unlike on an SDS gel where all samples are run within the same gel and transferred to the same membrane. Another disadvantage is that the kits they sell only have mouse and rabbit secondaries, so a secondary from any other organism will take some time to troubleshoot. Handling isn't overly complicated, and the Simple Wes definitely saves you time.
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