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Pippuri
Member since: Jul 12, 2009
From: Georgia, United States
Status: Protein Chemistry & Glycobiology Moderator
My points: 1033    what's this
Name: [Privacy]
 


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Feb 15, 2012    11:56 AM

Replied to topic in Analytical Chemistry forum   mass spectrometry

ghazaleh842000, I am not sure I understand fully your question. At the full ms level, there is no special "look" of each species. But if your full ms has multiple species on one spectrum, you should be able to back calculate for all of them share the same value of M, but vary in n value ...

Feb 15, 2012    07:07 AM

Replied to topic in Analytical Chemistry forum   mass spectrometry

Hi ghazaleh842000, For single charge species, the m/z value = [M+H], M being the mass of your target (says peptide, but can be anything), and H being mass of the hydrogen atom (1.0078). For doubly charge species, the m/z value = [M+2H]/2, M being the mass of your target (says peptide, but can ...

Feb 13, 2012    08:23 AM

Replied to topic in Protein Chemistry forum   mass spectrometry on Factor XIII

Hi ghazaleh842000, You need to collect MS/MS spectra (which contain information of b/y ion series specific for your peptides) for such assignment. I am not very familiar with MALDI system- for what I understand, since your sample is mixed with matrix prior to TOF-MS/MS analysis, there is no L ...

Feb 12, 2012    08:57 AM

Replied to topic in Protein Chemistry forum   mass spectrometry on Factor XIII

Hi ghazaleh842000, How long did you perform pepsin digestion? Have you tried to reduce the digestion time for B2?

Feb 12, 2012    08:57 AM

Replied to topic in Protein Chemistry forum   mass spectrometry on Factor XIII

Hi ghazaleh842000, How long did you perform pepsin digestion? Have you tried to reduce the digestion time for B2?

Feb 9, 2012    06:51 AM

Replied to topic in Protein Chemistry forum   mass spectrometry on Factor XIII

Hi ghazaleh842000, Can you pleasebe more specific about your approach? For instance, are you using MALDI or ESI based method? Are the sample analyzed on the whole protein level or have it been digested? How does the calibration/tuning conditions on the instrument? Cheers.

Feb 7, 2012    09:40 AM

Replied to topic in General forum   Isozyme electrophoresis

Hi AnjaliKrsih, It depends on how big difference are there between all these isozymes. If they have indeed very similar in size, 1D-PAGE with regular running condition is almost impossible to separate them. While increasing the electrophoresis time, you are indeed also increase the resolving powe ...

Feb 7, 2012    09:35 AM

Replied to topic in Biochemistry forum   calculating the kcat

HI namniow123, Divide your vmax (3.63 umole/min.ml) by 60 will transform its unit into umole/sec.ml. Then you are on the right track to use the equation for your kcat calculation. Good luck.

Jan 31, 2012    03:04 PM

Replied to topic in Protein Chemistry forum   His-Tag Recombinant Protein purification

Hi qpwoei4756, I suggest you to pass your protein through a second column, preferentially a size-exclusion chromatography. It should give you a much cleaner sample. Good luck.

Jan 31, 2012    10:58 AM

Replied to topic in Protein Detection (Western blots, gels, IP) forum   Question regarding Quantity One for semiquantification of western

Hi Isadora, I am still baffled by the negative value, but one thing I "sort of" figure out is to acquire the image under the UV light instead of the under the white light mode- that seems to somehow reduce the chance of getting negative value.... For the situation in which I couldn' ...

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