Does any one have the Bradford method for determination of protein in bacterial cell culture?
BCA protein quantification method is more accurate. Bradford method can be used also. But using bradford kit (i.e. sigma ) is better than preparing the Bradford reagent in the lab. Preparing fresh standard is also good to get better result everytime.
Thanks Dr. Haque for your answer. Could you attach the reference of BCA protein quantification method.
Thanks
The Bradford reagent can be easier and cheaper to use especially if you are testing culture supernatant. Be advised, the BCA kits can have significant interference with culture media. I haven't run into interference issues using the Bradford reagent.
Thanks Dr. Mann for your answer. Actually I am testing culture supernatant. By the way, can measuring protein in culture supernatant represent the bacterial growth?
Dr. Hayakawa,
Thanks for your answer. Actually the spectrophotometric-dependent analytical methods are more practical than HPLC, as spectrophotometer is present in nearly all labs, at least here in my country, I do not know the case in Japan.
Any way I will try your methods, thanks
Thank you for your kind answer. I surely agree that HPLC system may be a high price machine, but it is not so high as compared to such expensive machine as MALDI-TOF-MS.
Thus, I recently have used the rental HPLC system from a rental company of the Orix Co., Japan; i.e., rental fee was about 120,000 yen or 1,000 USD per month.
Short Diol-type SEC column (Develosil 50 Diol-5 35 x 8 mm I.D., 5 nm pore-diameter, 5 micro-meter particle size) was purchased from the Nomura Chemical Co., Seto-city, Aichi, Japan, for about 40,000 yen or 320 USD.
URL of the Nomura Co. is as follows,
http://www.develosil.net/english/index.html
Thanks Dr Hayakawa. It really appears cheaper if I use your idea for renting the HPLC system, but I have a question, is it the same price out of Japan?
I searched for ORIX rentec Co., and I found that Thailand is most near to your country.
http://www.orix.co.th/business-unit/test-measurament-equipment.asp
However, similar rental company may be present in your country (Egypt).
Please, ask the rental price to the adequate rental company.
I am sad to inform you the important issue that only the non-ionic detergent of
polyethylene-glycol 1000 monocetylether (PGC) of Wako (Osaka, Japan) can give a linear calibration line inserting to zero. But, Brij-58 (PGC) of Sigma gives a linear calibration line not inserting to zero; i.e., two points calibration is required. This is
possibly due to the differences in the molecular-weight distributions of PGCs; i.e., Brij-58 may have a wider range of molecular-weight (Mr) distribution as compared to Wako's PGC, and PGC of European product may be OK.
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