UltraflextremeTOF from Brucker is the MALDI-TOF instrument and matrix Sinapinic acid (10 mg/ml), TA 50(50:50 ACN: H2O 0.1% TFA ) was used to record higher molecular weight protein like BSA, but the mass spectra do not look good.
Hi Reddy,
if you intend to detect proteins in the mass range of 60kDa, I would increase the SA concentration up to 40 mg/mL, and use a sandwhich prep (Bogdan Munteanu et al., Anal. Bioanal. Chem., 2012.). You need also to optimize your parameters on the Ultraflex e.g. delay time go up to 600 ns (need to be optimized).Further you would need to optimze the voltages on your source.
I would use BSA as a standard and optimze the parameters on it. With an Ultraflextreme you can detect Antibodies with a "nice" resolution.
best regards
Bogdan
Hi Neelesh,
It's also helpful to desalt and concentrate the protein by C4 ZipTip. They are very helpful to give better spectrum with good resolution. Also as Bogdan said prepare double layered matrix. Apply 1 uL of saturated SA on the plate. Let it completely dry under RT condition (no fast drying). Then mix your desalted sample in dilution series with SA dissolved in TA30 and apply 1 uL. This might work better for you! ALl the best
Hello
Other little idea is to increase the number of spectra collected and to set manually acquisition, so you could chose the areas where you see better response and adjusting the laser power based in better crystallization.
Hello,
I also suggest you to optimize Lens voltage parameter of your Ultraflex source - should be increased with higher mass range, to 9 or 10 kV to BSA mass range.
Remember that you will be working in TOF linear mode, so your resolution won´t be high (about 5000 FWHM at 1000 m/z).
In our assays we got 3000 FWHM resolution for BSA, using the good tips suggested above by Munteanu and Palei.
Good luck.
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