QuicKey Human RBP4(Retinol Binding Protein 4, Plasma) ELISA Kit

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    • ELISA-Elabscience
    • ELISA-Elabscience
    • ELISA-Elabscience
    • ELISA-Elabscience
    • ELISA-Elabscience
    • ELISA-Elabscience

      Catalog number:E-TSEL-H0006

      Size:
      • 96T
      Qty:
      - +
      Price: $390

      Reactivity: Human

      Detection Range: 3.13~100 ng/mL

      Sensitivity: 1.98 ng/mL

      Lead Time: 7~10 daysWelcome to order from local distributors.

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      QuicKey Series

      Get more sensitive and precise results with saving at least 1h comparing to traditional ELISA Kits. The new developed technology in house will help to accelerate your science research in a more efficient way.


      Intended use

      This ELISA kit applies to the in vitro quantitative determination of Human RBP4 concentrations in serum, plasma, urine. Please consult technical support for the applicability if other biological fluids need to be tested.


      Background

      Retinol binding protein 4(RBP4), previously called retinol binding protein(RBP), is a transporter protein. This protein belongs to the lipocalin family and is the specific carrier for retinol (vitamin A alcohol) in the blood [1]. It delivers retinol from the liver stores to the peripheral tissues. In plasma, the RBP-retinol complex interacts with transthyretin which prevents its loss by filtration through the kidney glomeruli. A deficiency of vitamin A, blocks secretion of the binding protein post translationally and results in defective delivery and supply to the epidermal cells.RBP4 seems to be correlated with cardio metabolic markers in inflammatory chronic diseases, including obesity, type 2 diabetes, metabolic syndrome, and cardiovascular diseases (CVDs) [2].


      1. Rask L, Anundi H, Fohlman J, et al. The complete amino acid sequence of human serum retinol-binding protein. [J]. Upsala Journal of Medical Sciences, 1987, 92(2):115.

      2. Bobbert P, Weithäuser A, Andres J, et al. Increased plasma retinol binding protein 4 levels in patients with inflammatory cardiomyopathy [J]. European Journal of Heart Failure, 2014, 11(12):1163-1168.


      Test principle

      This ELISA kit uses the Sandwich-ELISA principle. The micro ELISA plate provided in this kit has been pre-coated with an antibody specific to Human RBP4. Samples (or Standards) and biotinylated detection antibody specific for Human RBP4 are added to the micro ELISA plate wells. Human RBP4 would combined with the specific antibody. Then Avidin-Horseradish Peroxidase (HRP) conjugate are added successively to each micro plate well and incubated. Free components are washed away. The substrate solution is added to each well. Only those wells that contain Human RBP4, biotinylated detection antibody and Avidin-HRP conjugate will appear blue in color. The enzyme-substrate reaction is terminated by the addition of stop solution and the color turns yellow. The optical density (OD) is measured spectrophotometrically at a wavelength of 450 ± 2 nm. The OD value is proportional to the concentration of Human RBP4. You can calculate the concentration of Human RBP4 in the samples by comparing the OD of the samples to the standard curve.

      Assay type

      Sandwich-ELISA

      Format

      96T

      Assay time

      2.5h

      Reactivity

      Human

      Detection method

      Colormetric

      Detection range

      3.13-100 ng/mL

      Sensitivity

      1.98 ng/mL

      Sample volume

      100μL

      Sample type

      Serum, Plasma, Urine


      Specificity

      This kit recognizes Human RNP4 in samples. No significant cross-reactivity or interference between RBP4 and analogues was observed.


      Typical data

      As the OD values of the standard curve may vary according to the conditions of  the actual assay performance (e.g. operator, pipetting technique, washing technique or temperature effects), the operator should establish a standard curve for each test. Typical standard curve and data is provided below for reference only.


      Concentration(ng/mL)

      100

      50

      25

      12.5

      6.25

      3.13

      0

      OD

      2.703

      1.717

      0.835

      0.362

      0.167

      0.103

      0.068

      Corrected OD

      2.635

      1.649

      0.767

      0.294

      0.099

      0.035

      -



      Sample values

      Serum/Plasma/Urine–Samples from apparently healthy volunteers were evaluated for the presence of Human RBP4 in this assay.

      Sample Type

      Source

      Range

      Dilution Factor

      Serum (n=14)

      Healthy human

      13.7-52.07μg/mL

      500-10000

      EDTA plasma (n=14)

      Healthy human

      15.44-57.49μg/mL

      500-10000

      Urine (n=12)

      Healthy human

      16.84-240.74ng/mL

      1-40


      Precision

      Intra-assay Precision (Precision within an assay): 3 samples with low, mid range and high level Human RBP4 were tested 20 times on one plate, respectively.

      Inter-assay Precision (Precision between assays): 3 samples with low, mid range and high level Human RBP4 were tested on 3 different plates,20 replicates in each plate.

       

      Intra-assay Precision

      Inter-assay Precision

      Sample

      1

      2

      3

      1

      2

      3

      n

      20

      20

      20

      20

      20

      20

      Mean(ng/mL)

      4.12

      11.24

      44.68

      4.56

      11.35

      48.06

      Standard deviation

      0.22

      0.51

      2.1

      0.27

      0.53

      2.52

      CV (%)

      5.34

      4.54

      4.7

      5.92

      4.67

      5.24



      Recovery

      The recovery of Human RBP4 spiked at three different levels in samples throughout the range of the assay was evaluated in various matrices.

      Sample Type

      Range (%)

      Average Recovery (%)

      Serum (n=14)

      81-108

      88

      EDTA plasma (n=14)

      82-120

      98

      Urinen=12

      80-119

      92


      Linearity

      Samples were spiked with high concentrations of Human RBP4 and diluted with Reference Standard & Sample Diluent to produce samples with values within the range of the assay.

       

       

      Serum (n=4)

      EDTA  plasma (n=4)

      Urine(n=4)

      1:2

      Range (%)

      81-113

      88-99

      87-105

      Average (%)

      95

      93

      96

      1:4

      Range (%)

      86-107

      82-105

      90-102

      Average (%)

      100

      92

      98

      1:8

      Range (%)

      91-97

      86-118

      86-110

      Average (%)

      94

      97

      95

      1:16

      Range (%)

      112-118

      80-120

      84-105

      Average (%)

      115

      93

      95


      Kit components & Storage

      An unopened kit can be stored at 2-8 for six months. After test, the unused wells and reagents should be stored according to the table below.

      Item

      Specifications

      Storage conditions after test

      Micro ELISA Plate (Dismountable)

      8 wells ×12 strips

      2-8, 1 months

      Reference Standard

      2 vials

      Discard unused reconstituted standard and dilutions

      Reference Standard & Sample Diluent

      1 vial, 20 mL

      2-8

      Biotinylated Detection Ab Working Solution

      1 vial, 12 mL

      HRP Conjugate Diluent

      1 vial, 14 mL

      Concentrated Wash Buffer (25×)

      1 vial, 30 mL

      Concentrated HRP Conjugate (100×)

      1 vial, 120 μL

      2-8(Protect from light)

      Substrate Reagent

      1 vial, 10 mL

      Stop Solution

      1 vial, 10 mL

      2-8

      Plate Sealer

      5 pieces

       

      Product Description

      1 copy

      Certificate of Analysis

      1 copy

      Note: All reagent bottle caps must be tightened to prevent evaporation and microbial pollution.

      The volume of reagents in partial shipments is a little more than the volume marked on the label, please use accurate measuring equipment instead of directly pouring into the vial(s).


      Other supplies required

      Microplate reader with 450 nm wavelength filter

      High-precision transfer pipette, EP tubes and disposable pipette tips

      37℃ Incubator

      Deionized or distilled water

      Absorbent paper

      Loading slot for Wash Buffer


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