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Stock availability
In Stock
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Kit s description
Colorimetric measurement of RBP levels in urine
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Protein target
Retinol Binding Protein Urinary
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Scientific context
Retinol binding protein (RBP) is from a family of structurally related proteins that bind small hydrophobic molecules such as bile pigments, steroids, odorants, etc (1). RBP is a 21 kDa highly conserved, single-chain glycoprotein, consisting of 182 amino acids with 3 disulfide bonds, that has a hydrophobic pocket which binds retinol (vitamin A). RBP binds retinol in a 1:1 stoichiometry, which serves to not only solubilize retinol but also protect it from oxidation. When in serum, the majority of RBP bound with retinol is reversibly complexed with transthyretin (prealbumin) (2,3). This complex then transports retinol to specific receptors of various tissues in the body. Vitamin A status is reflected by serum concentration as it is hemostatically controlled and does not fall until stores are dramatically reduced (4,5). RBP has also been shown to be a useful marker for renal function (6) as it is totally filtered by the glomeruli and reabsorbed by proximal tubules (7). This has made urinary RBP (uRBP) a tool to study renal function in heart (8) or kidney (9) transplant recipients, type 1 and 2 diabetics (10), and in people exposed to uranium from mining operations (11). Measurement of uRBP levels has also been useful in detection and characterization of diseases including hypertension (12) and certain cancers (13,14).
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Primary research area
Cell Signaling, Cardiovascular System
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Category
Assay EIA Kits
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Other name
Cellular retinol binding protein EIA Kit, CRBP EIA Kit, CRBP EIA Kit, CRBP1 EIA Kit, CRBP2 EIA Kit, RBP1 EIA Kit, RBP2 EIA Kit, RBP4 EIA Kit, RBPC EIA Kit, Retinol binding protein 1 EIA Kit, Retinol binding protein 2 EIA Kit, Retinol binding protein 4 EIA Kit
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Brand name
StressXpress®
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Detection system
Colorimetric Assay
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Assay format
Sandwich EIA (Enzyme Immunoassay)
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Main intended usage
EIA kit used to measure RBP levels in urine samples.
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Brief protocol
The Retinol Binding Protein Urinary EIA Kit is designed to quantitatively measure RBP present in urine samples. A RBP standard is provided to generate a standard curve for the assay and all samples should be read off the standard curve. Standards or diluted samples are pipetted into a clear microtiter plate coated with an antibody to capture rabbit antibodies. A RBP-peroxidase conjugate is added to the standards and samples in the wells. The binding reaction is initiated by the addition of the RBP polyclonal antibody to each well. After an hour incubation the plate is washed and substrate is added. The substrate reacts with the bound RBP-peroxidase conjugate. After a short incubation, the reaction is stopped and the intensity of the generated color is detected in a microtiter plate reader capable of measuring 450 nm wavelength. The concentration of the RBP in the sample is calculated, after making a suitable correction for the dilution of the sample, using software available with most plate readers.
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Kit contents
Clear 96 well Plate, RBP Standard, StressXpress® RBP Antibody, StressXpress® RBP-Peroxidase Conjugate, Assay Buffer, Wash Buffer Concentrate, TMB Substrate, Stop Solution, Plate Sealer
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Assay precision
Intra Assay Precision: Four human urine samples were diluted 1:2 with Assay Buffer and run in replicates of 8 in an assay. The mean and precision of the calculated RBP concentrations were: Sample 1- 14.8 ng/mL, 4.5% CV Sample 2- 28.0 ng/mL, 7.5% CV Sample 3- 53.5 ng/mL, 7.3% CV Sample 4- 323.7 ng/mL, 2.1% CV Inter Assay Precision: Four human urine samples were diluted 1:2 with Assay Buffer and run in duplicates in twenty-one assays run over multiple days by three operators. The mean and precision of the calculated RBP concentrations were: Sample 1- 14.4 ng/mL, 9.1% CV Sample 2- 28.0 ng/mL, 9.2% CV Sample 3- 50.8 ng/mL, 8.1% CV Sample 4- 309.2 ng/mL, 14.0% CV
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Assay cross reactivity
Human, Rat, Dog, Monkey
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Samples to be used with this kit
Urine
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Maximum samples to be used with this kit
41 samples in duplicate
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Assay duration
90 Minutes
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Kit s sensitivity
2,90 ng/ml
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Assay detection limit
3,906 - 1000 ng/ml
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Storage recommendations
4°C
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Shipping recommendations
Blue Ice
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Bibliography
1. Blaner WS. Endocr Rev. 1989 Aug;10(3):308–16. 2. Wolf G. Physiol. Rev. 1984 Jul;64(3):873-937. 3. Petersen PA. J. Biol. Chem. 1971, 246:34-43. 4. Goodman DS, Blaner WS. in The Retinoids, eds. Sporn MB, Roberts AB, Goodman DS, (Orlando: Academic Press, 1984) vol.2, 1-39. 5. Olson, JA. Modern Nutrition in Health and Disease, eds. Shils ME, Olson JA, Shike M, (Philadelphia: Lea & Febiger, 1994) 8th ed. 287–307. 6. Peterson PA, and Berggård I. J. Biol. Chem. 1971 246:25-33. 7. Bernard AM, and Lauwerys RR. Clin. Chem. 1981 27:1781-2. 8. Camara NO, Matos AC, Rodrigues DA et al. The Lancet 2001 357: 856-857. 9. Hosaka B, Park SI, Felipe CR, Garcia RG, Machado PG, Pereira AB, Tedesco-Silva H and Medina-Pestana JO. Transplant Proc. 2003 35:1341-1343. 10. Abahusain MA, Wright J, Dickerson JWT and de Vol EB. Am J Clin Nutr. 2004;79:218-25. 11. Wyatt SA, Reitz LV, Croley TR, Hawkins D, Barrett E, McKeown A, Powell N, West A, Hamner T and Royster MO. J Exp Sci Environ Epidem. 2008 18:59-75. 12. Bang LE, Holm J and Svendsen TL. Am. J. Hypertens 1996 9:1024-1028. 13. Moise AR, Noy N, Palczewski K, Blaner WS. Biochemistry 2007 9:1024-1028. 14. Gavrilov V, Yermiahu T, Gorodischer R. Kidney Int. 2006 Jan;69(1):173-7.
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Release date
17-Sep-2014
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PubMed number
Refer to PubMed
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Tested applications
No data available / currently testing
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Tested reactivity
No data available / currently testing
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Resources available upon request
Kit Booklet, MSDS
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Product image link
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Product image legend
Typical Standard Curve for the Retinol Binding Protein Urinary EIA Kit (Enzyme Immunoassay) StressXpress® - SKT-218. Assay Type: Sandwich EIA. Detection Method: Colorimetric Assay. Assay Range: 3.906 - 1000 ng/ml. | Linearity was determined by taking two human urine samples diluted 1:2, one with a low diluted RBP level of 14.5 ng/mL and one with a higher diluted level of 299.2 ng/mL and mixing them in given ratios. The measured concentrations were compared to the expected values based on the ratios used. Graph of the Typical Standard Curve for the Retinol Binding Protein Urinary EIA Kit StressXpress - SKT-218 | Graph of the Linearity Recovery for the Retinol Binding Protein Urinary EIA Kit StressXpress - SKT-218
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Warnings
Non-hazardous
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Country of production
Canada
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Total weight kg
2
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Net weight g
300
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Description
This ELISA test kit or EIA Enzyme immuno assay is an enzyme linked immuno assay supplied as coated 96 well plates with antigen or antibody and needs to be stored at +4°C.
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Properties
MeSH E05 478 566 350 170 Enzyme-Linked Immunosorbent Assay
