northernlad2690

I'm using it for the calculation of an enzyme assay when calculating the enzyme activity of a recombinant protein. It indicates umol of substrate catalysed per minute, per umol of recombinant enzyme added to the assay. - first time I've ever used this specific methods for calculating enzyme activity but is being used prior to calculating katal.

Hello, I'm just wanting someone to give me a little bit of guidance on the following conversion, i'm trying to convert my reading of umol/min/umol to umol/min/mg- i think i have the correct answer but i need my method validated if someone could possibly help? i have a spectrophotometric assay of a purified recombinant enzyme, Spy057, against p-nitrophenyl N-acetyl-β-glucosaminide substrate (total reaction volume, 2 mL; volume of Spy05783 added, 50 µL): - I have calculated the umol/min/umol using beer lamberts law to 289.63 - the Mr of spy057 is 43507g/mol - i then calculated that there is 43.507mg/umol - there is 1.899 x 10E-4 umol added reaction volume so using this i then calculated 43.507x(1.899E-4) = 8.272 x 10E-3 using data obtained previously i knew that the umol/min of p-nitrophenol was 0.055 so this was input into the final equation 0.055/8.272 x 10E-3 giving me 6.657umol/min/mg does this look correct, ive done everything to try and find a protocol but cant find anything

Hello, I'm just wanting someone to give me a little bit of guidance on the following conversion, i'm trying to convert my reading of umol/min/umol to umol/min/mg- i think i have the correct answer but i need my method validated if someone could possibly help? i have a spectrophotometric assay of a purified recombinant enzyme, Spy057, against p-nitrophenyl N-acetyl-β-glucosaminide substrate (total reaction volume, 2 mL; volume of Spy05783 added, 50 µL): - I have calculated the umol/min/umol using beer lamberts law to 289.63 - the Mr of spy057 is 43507g/mol - i then calculated that there is 43.507mg/umol - there is 1.899 x 10E-4 umol added reaction volume so using this i then calculated 43.507x(1.899E-4) = 8.272 x 10E-3 using data obtained previously i knew that the umol/min of p-nitrophenol was 0.055 so this was input into the final equation 0.055/8.272 x 10E-3 giving me 6.657umol/min/mg does this look correct, ive done everything to try and find a protocol but cant find anything

i wonder if anyone could offer some subtle insight, I have a set of standards of substance X with an appropriate standard curve constructed from this info. The sample of unknown concentration is diluted 10X and thus has produced a much smaller absorbance reading as expected, but I'm stuck on how to correct for this dilution as i'm not sure whether to; A) multiply the absorbance reading by 10 or B) multiply by 10 the figure from the point on the graph where the diluted absorbance reading is.

i wonder if anyone could offer some subtle insight, I have a set of standards of substance X with an appropriate standard curve constructed from this info. The sample of unknown concentration is diluted 10X and thus has produced a much smaller absorbance reading as expected, but I'm stuck on how to correct for this dilution as i'm not sure whether to; A) multiply the absorbance reading by 10 or B) multiply by 10 the figure from the point on the graph where the diluted absorbance reading is.

thanks for the reply, I got there eventually.

im assuming you are looking at Nickel affinity chromatography.

What is the general principle of PCR-SSP? I get it uses sequence-specific primers to bind only to selected alleles but isn't this the principle in all PCR techniques?