Abstract：
:The retinal pigment epithelium (RPE) is essential for maintaining the health of the neural retina. RPE cell dysfunction plays a critical role in many common blinding diseases including age-related macular degeneration (AMD), diabetic retinopathy, retinal dystrophies. Mouse models of ocular disease are commonly used to study these blinding diseases. Since isolating the RPE from the choroid has been challenging, most techniques separate the RPE from the retina, but not the choroid. As a result, the protein signature actually represents a heterogeneous population of cells that may not accurately represent the RPE response. Herein, we describe a method for separating proteins from the RPE that is free from retinal and choroidal contamination. After removing the anterior segment and retina from enucleated mouse eyes, protein from the RPE was extracted separately from the choroid by incubating the posterior eyecup with a protein lysis buffer for 10 min. Western blot analysis identified RPE65, an RPE specific protein in the RPE lysates, but not in choroidal lysates. The RPE lysates were devoid of rhodopsin and collagen VI, which are abundant in the retina and choroid, respectively. This technique will be very helpful for measuring the protein signal from the RPE without retinal or choroidal contamination.
journal_name
Exp Eye Resjournal_title
Experimental eye researchauthors
Wei H,Xun Z,Granado H,Wu A,Handa JTdoi
10.1016/j.exer.2015.09.015subject
Has Abstractpub_date
2016-04-01 00:00:00pages
450-455eissn
0014-4835issn
1096-0007pii
S0014-4835(15)30036-1journal_volume
145pub_type
杂志文章abstract：:Tears serve as a viable diagnostic fluid with advantages including less invasive sample to collect and less complex to prepare for analysis. Several water-soluble and fat-soluble vitamins were detected and quantified in human tears and compared with blood serum levels. Samples from 15 family pairs, each pair consistin...
journal_title：Experimental eye research
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journal_title：Experimental eye research
pub_type： 杂志文章
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更新日期：1997-03-01 00:00:00
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更新日期：2004-03-01 00:00:00
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journal_title：Experimental eye research
pub_type： 杂志文章
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更新日期：1997-04-01 00:00:00
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更新日期：2014-09-01 00:00:00
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更新日期：1989-11-01 00:00:00
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更新日期：2020-04-01 00:00:00
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pub_type： 杂志文章,评审
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journal_title：Experimental eye research
pub_type： 杂志文章
doi：10.1006/exer.1994.1003
更新日期：1994-02-01 00:00:00
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journal_title：Experimental eye research
pub_type： 杂志文章
doi：10.1016/s0014-4835(85)80025-7
更新日期：1985-09-01 00:00:00
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journal_title：Experimental eye research
pub_type： 杂志文章
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更新日期：2000-04-01 00:00:00
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pub_type： 杂志文章
doi：10.1016/s0014-4835(87)80097-0
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pub_type： 杂志文章,meta分析,评审
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更新日期：2015-10-01 00:00:00
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journal_title：Experimental eye research
pub_type： 杂志文章
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pub_type： 杂志文章
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更新日期：1983-07-01 00:00:00
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journal_title：Experimental eye research
pub_type： 杂志文章
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更新日期：1984-08-01 00:00:00
abstract：:The lysosomal proteases cathepsin B and dipeptidyl peptidase II (DPP II) were identified biochemically in the supernatant fraction of rat and guinea-pig lens homogenates, using as substrates CBZ-Arg-Arg-4-methylcoumarin-7-amide (AMC) (5 microM, pH 6.0, 37 degrees C for cathepsin B, and Lys-Ala-AMC or Lys-Pro-AMC (5 mi...
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pub_type： 杂志文章
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更新日期：1987-06-01 00:00:00
