Has anybody tried western blotting related buffers, antibodies etc from Sigma? How do they compare with products from abcam?
I've used a monoclonal antibody from sigma (anti phospho ERK). It is very good, I used 1:1000 dillution and it worked all the time. However antibody quality depends on the used clone and other parameters. You should check the literature for your specific antibody of choice. For buffers and reagents Sigma offers high quality solutions so I believe you wouldn't have any problems with these kind of products.
Thank you Andre. That helps. Also Do you think pre-cast PAGE gels are worth the cost?
Honestly, It depends on your experiment and the result you are looking for. I'm from Brazil so money is an important issue. Pre cast gels are a little expensive here so I would perform my padronization (antibody and sample titration, time lapse determination) using normal gels. Once I had established all parameters (and if I wans't able to get a nice home made gel for publicatio) then I would go for the pre cast one. The salesman alway say these gels have expiration dates but if stored properly they last way longer than expected. If I were you I would only buy pre cast gel if absolutely necessary and if funding is available. I prefer spending a little extra in a higher quality antibody than in buffers and pre cast gels. If you use a good blocking agent (I've used non fat dry milk), a good BSA (sigma BSA is very good) and always sterilize your buffers by filtration you wont't have any problem. But again this is the perspective from someone who would always have to consider the financial issue.
thank you for the advice Andre. All these points are very useful for me. Money is an important consideration in Pakistan as well
With Sigma, you'll be paying a lot extra for the brand name and sales people's salaries. I for one can't see their products being any better than what you'd get from smaller companies for half the price. Genscript for example has very cost effective pre-cast gels (if you buy a lot of them, due to shipping). Antibodies I'd look at biolegend, ebioscience. Also use price-comparison tools like Quartzy, p212121. Those are going to kill big companies' margins.
I agree with John. Sigma tends to charge top dollars for similar quality reagents that are sometimes available cheaply from other companies. Tania, to be honest with you, I see no reason in buying buffers or pre-cast gels from commercial sources unless absolutely neccessary. Making reagents is part of our job and lots of fun. Being an avid angler I find as much joy in preparing my gear as I do in reeling out the fish. Reagents that can be prepared cost effectively in the lab should be made in the lab. As far as antibodies are concerned, try to purchase antibodies from sources that provide images of western blots. Some times they dont but in that case antibodies that have been referenced in publications are your best bet. We make our own antibodies so this does not bother me a lot.
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