Immunoquantitation of aldose reductase in human tissues

Analytical Biochemistry
C E Grimshaw, E J Mathur

Abstract

Rabbit antibodies raised against bovine kidney aldose reductase (ALR2) were shown to be monospecific for human ALR2 by Western blot analysis of human muscle homogenates. The human enzyme was detected, by reaction with the antiserum (alpha-BKALR2), in homogenates of adrenal gland, muscle, lens, brain, testes, kidney, and placenta, but not in erythrocytes or leukocytes. The amount of enzyme in each tissue was determined by densitometric analysis of autoradiographs of Western blots probed with alpha-BKALR2 and [125I]protein A. Standard curves of radiographic intensity versus amount of purified human muscle ALR2 were linear in the 20 to 200-ng range; a similar sensitivity was seen in tissue homogenates containing up to 675 micrograms total protein. The results presented here for the ALR2 level in human tissues (adrenal greater than muscle greater than lens approximately brain approximately testes greater than kidney approximately placenta) are in agreement with literature values for those tissues from which the enzyme has previously been purified. A notable exception was the absence of detectable ALR2 in human erythrocytes. A quantitative comparison of immunoradiographic response showed that bovine kidney ALR2 was about sevenfold m...Continue Reading

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Citations

Dec 8, 1993·Biochimica Et Biophysica Acta·B RobinsonD L Vander Jagt
Jun 1, 1996·Diabetes Research and Clinical Practice·E Kicic, T N Palmer
Jul 1, 2010·Molekuliarnaia biologiia·E S KropotovaT D Mashkova
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Aug 26, 1998·The Journal of Clinical Endocrinology and Metabolism·V O ShahP G Zager
Mar 27, 2007·The Biochemical Journal·Matthew SpiteSanjay Srivastava

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