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amount of proteins in urine


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13 replies to this topic

#1 ujla80

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Posted 05 July 2009 - 09:10 AM

hi all

can anybody tell me the amount of proteins excreted in urine of normal beings? is lowry a good method for detecting urinary proteins?
help me please.... :P
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#2 HomeBrew

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Posted 05 July 2009 - 12:52 PM

Accoring to here, the normal range of total protein on urinalysis is 6.3-8.2 gm/dL. But, according to here, the normal value is "negative". I suppose it depends on the sensitivity of the assay used.

BTW, 63 - 82 g/L seems pretty high to me...

#3 hobglobin

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Posted 05 July 2009 - 12:55 PM

Accoring to here, the normal range of total protein on urinalysis is 6.3-8.2 gm/dL. But, according to here, the normal value is "negative". I suppose it depends on the sensitivity of the assay used.

But the in first link also gives negative-trace Protein in Urinalysis...I guess normally a test strip (showing different intensity of lilac staining) is used for routine analysis...don't know how sensitive this method is.
But after stress such as sports etc. the protein concentration is raised at least temporarily.

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...except casandra's that belong to the funniest, most interesting and imaginative (or over-imaginative?) ones, I suppose.


#4 ujla80

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Posted 05 July 2009 - 06:12 PM

thanx ;)

i am looking for proteinuria in mice urine samples. with lowry i get very high protein levels 160mg/dl-3000mg/dl in controls(normal healthy mice)...whereas when i do strip test i get lower amounts i.e trace-100mg/dl....what could be the reason of such high values with lowry?
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#5 Gerard

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Posted 06 July 2009 - 05:45 AM

Is it possible your using a teststrip that only detects albumin? :)
Ockham's razor
Pluralitas non est ponenda sine necessitate
-- "You must assume no plural without necessity".

#6 mdfenko

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Posted 06 July 2009 - 07:57 AM

you are probably above the useful range of the lowry assay.

clinical labs use the biuret protein assay. it is useful for greater amounts of protein than lowry.
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#7 ujla80

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Posted 06 July 2009 - 08:16 AM

the test strip is more sensitive to albumin than other proteins...

m going to use biurett method and see the results. :)
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#8 DRT

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Posted 06 July 2009 - 07:21 PM

Wouldn’t the ammonium/urea type compounds in urine cause problems for both the Biuret and Lowry assays or are the concentrations too low to worry about?
Is it possible that the pH was different between the standards and samples for the Lowry assay?

#9 mdfenko

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Posted 07 July 2009 - 08:02 AM

Wouldn't the ammonium/urea type compounds in urine cause problems for both the Biuret and Lowry assays or are the concentrations too low to worry about?
Is it possible that the pH was different between the standards and samples for the Lowry assay?

urea won't significantly affect the assay. biuret has a peptide-like bond which is detected by the assay, urea doesn't (although, there may be some biuret in the urea and that will be detected).

the pH of the samples won't have a significant effect on the lowry (or biuret). the reagent has enough naoh to maintain a basic environment.
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#10 sgt4boston

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Posted 17 August 2009 - 11:27 AM

Are you looking for particular proteins? Gammopathies?

#11 chdd

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Posted 20 August 2009 - 12:26 PM

i had used Bradford method(Coomassie brilliant blue G-250) to assay proteins in urine,it seems good.

#12 ujla80

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Posted 22 August 2009 - 10:38 PM

no not specific proteins. i am just looking for proteinuria...


Are you looking for particular proteins? Gammopathies?


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#13 ujla80

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Posted 22 August 2009 - 10:41 PM

hi :lol:

did u dilute the urine sample if yes then how much dilution?


i had used Bradford method(Coomassie brilliant blue G-250) to assay proteins in urine,it seems good.


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#14 Zagami Francesco

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Posted 08 June 2015 - 10:46 AM

I measure microalbuminuria by means a screening spot test (SST) on cellulose acetate employing 2 μl of random fresh emitted (<2 h) or stored at 4 °C urine specimens that are spotted on dry cellulose acetate and allow to stay for 2-3 min until spot area is completely dry. Immerse cellulose acetate film in dye CBB R-250 solution (dissolve in glass amber bottle, 100 mg of Coomassie R-250 in 22.5 ml of methanol, mixing from time to time for inversion for then minutes, and add 5.0 ml acetic acid, mix as above, and finally add 22.5 ml of dH2O, mix as above and allows to rest for at least 5 hrs before use. Stable 1 year at room temperature) and incubate for 4 min. at RT. Draw and drain the film and to set it in destain solution (5% acetic acid aqueous solution) for 3 cycles of 10 min. Spot positive is underlined by different intensity blue color proportional to albuminuria concentration, while the uncolored spot area identified the specimen as normal value. Calibration is carried out using human serum albumin saline solutions ranging from 20 to 200 mg/L. Starting from albumin standard 60 g/L (60000 mg/L) dilute 1:100 (100 μl + 9.9 ml) with normal saline solution (NSS - 0.9% NaCl) obtaining standard 600 mg/L and from this made decreasing serial dilution always in normal saline as below indicated :
300 mg/L : 500 μl 600 mg/L ST + 500 μl NSS; 200 mg/L : 400 μl 300 mg/L + 200 μl NSS; 150 mg/L : 300 μl 200 mg/L ST + 100 μl NSS; 100 mg/L : 300 μl 150 mg/L + 150 μl NSS; 75 mg/L : 300 μl 100 mg/L + 100 μl NSS; 50 mg/L : 200 μl 75 mg/L ST + 100 μl NSS; 30 mg/L : 150 μl 50 mg/L + 100 μl NSS; 20 mg/L : 100 μl 30 mg/L + 50 μl NSS. For test screening compare the color intensity spot of the sample with that of the serial calibration range.




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