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Pippuri
Member since: Jul 12, 2009
From: Georgia, United States
Status: Protein Chemistry & Glycobiology Moderator
My points: 1069    what's this
Name: [Privacy]
 


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Mar 24, 2015    12:48 AM

Replied to topic in Biochemistry forum   Determining Enzyme Kinetics

Tara-linth, To plot Michealis Menten, the x-axis is substrate concentration and y-axis is velocity, Hence, you need to calculate your enzyme velocity (rate at which product is formed). If you have a standard curve where you know the absorbance of your product, you should be able to back calcu ...

Mar 18, 2015    04:32 PM

Replied to topic in DNA (PCR, Real Time PCR) forum   Primers

one additional side note to what creativeproteomics has touched on- I preferred to prepare the working solution of primers with ddH2O. Since the DNA polymerase reaction is Mg-dependent, by doing the 1/10 dilution with ddH2O can ensure the amount of EDTA in the final PCR reaction become insignific ...

Mar 4, 2015    04:46 PM

Replied to topic in Diagnostics (Molecular, Pathological) forum   DNA dilution

Hi jem, Since you start with 1000nmol per 1 ml (or 1000 nmol per 1000 ul), the concentration = 1 nmol per ul. Hence, by taking 5 ul from your stock, it will contain 5 nmole of your oligo.

Feb 12, 2015    07:28 PM

Replied to topic in Cell Culture and Tissue Culture forum   Cell loss during annexin staining/flow cytometry

Hi Sophie, what is the source of neutrophil you use in your experiment? Did you observe any clumps while handling them?

Feb 5, 2015    08:12 PM

Replied to topic in Cell Culture and Tissue Culture forum   does MG132 dissolve in culture media? pls im desperate for help

Hi Annafreud, There are couple possibilities- How did you prepare MG132? While MG132 is very stable in organic solvent, it is rather unstable in aqueous condition. It has to be prepare fresh in the aqueous solution (ie. media) on the same day of experiment. The cells that you are treating M ...

Feb 5, 2015    05:43 PM

Replied to topic in Cell Culture and Tissue Culture forum   does MG132 dissolve in culture media? pls im desperate for help

Hi Annafreud, It isn't clear to me the problem that you are encountering is either the irreproducible results or the cells are dying (or not dying). Can you please elaborate?

Feb 4, 2015    10:47 PM

Replied to topic in Biochemistry forum   Sequence specific pulldown of dsDNA

Hi Baconman, In theory, it should work. I guess it is just a matter of optimization, which requires patience of a saint! :-) Good luck.

Nov 26, 2014    08:06 PM

Replied to topic in Protein Detection (Western blots, gels, IP) forum   Detecting small MW protein via SDS-page / Western

Loulou34, How much methanol or ethanol you have in your transfer buffer?

Oct 20, 2014    05:26 PM

Replied to topic in Neuroscience forum   PC12 and voltage gated calcium channels

Hi Domino, You can use ω-Conotoxin MVIIC for blocking N and P/Q type Cav channel. Good luck.

Aug 28, 2014    04:36 PM

Replied to topic in Cancer forum   Western blot analysis

Hi Britjade91, What is the blot corresponding to the imageJ histogram? phospho-ERK? Does the blot give you one or two bands? Depends on the source of your sample, some time you see one band, some time you see two... If you are getting two bands and both of them are your target, then I would gr ...

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