sod assay

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banda1210
banda1210's picture
sod assay

HI every one.,
I have few doughts regarding sod assay.,
Why we need to expose the reaction mixture to flouresent lamp only? What if i use normal incandesent lamp and how the flouresent lamp would aid in the reaction to occur.,?
kindly suggent me-thank u

Satinder
Satinder's picture
fluoroscent lamp emits the

fluoroscent lamp emits the monochromatic wavelength in the fluoroscent range where as incandescent light uses heat driven light emission.

it is more regulated and uniform for the Former then compare to the incandescent light source.

banda1210
banda1210's picture
hi sir.,

hi sir.,
i have extracted the crude enzyme of sod ,cat,apx ., i'm following the giannopolitis and ries 1977 method to assay the particular enzymes., i did not understod how to quantify the protein., (1 unit of sod)., should i require all the three enzymes for the assay of sod, apx,cat i mean.,

           kindly suggest me sir.,
          Thank u .....

fatma_elzahraa
fatma_elzahraa's picture
dear,

dear,

to calculate the specific activity of sod,,, u should first construct standard curve of sod to determine its concentration at which 50% inhibition of pyrogallol auto-oxidation occurs...
then 1 unit of sod is equivalent to (that concentration at 50% inhibition)
so u can divide the concentration of sod u get in ur sample by that concentration equivalent to 50% inhibition then divide it again by the protein concentration in this sample and adjust the units to be U/mg protein

i hope u can understand me...

don't hesitate to contact if u need any further info.

best regards  

Vedant Kevlani
Vedant Kevlani's picture
SOD activity

Hellow mami performed SOD assay using method of mishra and fridovich, 1972Which is based on the ability of SOD to inhibit the auto-oxidation of epinephrine to adrenochrome at alkaline pHBriefly,thesupernatant(25 ml) is  added to a mixture of epinephrine(0.1mM) in carbonate buffer(pH10.2)in a totalvolume of 1ml and the formation of adrenochrome wasmeasured at 295nm.Readings were taken at 0 and 60 secNow i want to calculate SOD (u/mg protien) using standard curveCan you tell me how to prepare standard curve and how to calculate activity?Thanks 

Vedant Kevlani
Vedant Kevlani's picture
SOD activity

SOD activityHellow mami performed SOD assay using method of mishra and fridovich, 1972Which is based on the ability of SOD to inhibit the auto-oxidation of epinephrine to adrenochrome at alkaline pHBriefly,thesupernatant(25 ml) is  added to a mixture of epinephrine(0.1mM) in carbonate buffer(pH10.2)in a totalvolume of 1ml and the formation of adrenochrome wasmeasured at 295nm.Readings were taken at 0 and 60 secNow i want to calculate SOD (u/mg protien) using standard curveCan you tell me how to prepare standard curve and how to calculate activity?Thanks  

    rozakuku
    rozakuku's picture
    SIR I WANT TO FUL INFORMATION

    SIR I WANT TO FUL INFORMATION TO CALCULATE THE SOD ACTIVITY FROM BLOOD SAMPLE