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Analysis of Renal Function & Injury---RBP4

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Retinol Binding Protein 4(RBP4) belongs to the lipocalin family and is the specific carrier for retinol (vitamin A alcohol) in the blood. 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 posttranslationally and results in defective delivery and supply to the epidermal cells.


Retinol-binding protein 4 has recently been described as an adipokine that contributes to insulin resistance in the AG4KO mouse model. It is secreted by adipocytes, and can act as a signal to other cells, when there is a decrease in plasma glucose concentration.Mutations in the RBP4 gene have recently been linked to a form of autosomal dominant microphthalmia, anophthalmia, and coloboma (MAC) disease.


Anti-Human RBP4 Monoclonal Antibody

Mouse anti-Human RBP4 antibodyhas been derived from the mouse immunized with human RBP4. MouseAnti-Human RBP4 monoclonal antibodycan be used for detection of RBP4 in latex enhanced immune turbidimetry.

Properties

Specification

Target species

human

Host animal

Mice Balb/c

Cell line used for fusion

Sp2/0

Immunogen

Human Retinol Binding Protein 4

Purification method, Purity

Protein Gaffinity chromatography,>90%(SDS-PAGE)

Presentation

MAb solution in NaCl with 15 mM NaN3(pH 7.2)

Application

LETIA ect.

Catalog Number

CSB-DA002AmN①

CSB-DA002AmN②

 

Calibration Curve

RBP4 proteins specifically react with anti-RBP4 monoclonal antibodies precoated onto latex beads to form insoluble complexes, resulting in turbidity increasing, and then the increasing of absorbance is detected by automatic biochemical analyzer. The calibration curve was fitted according to the relationship between absorbance values and RBP4 concentrations (Fig.1). 



Fig.1 Calibration curve forRBP4in latex-enhanced turbidimetric immunoassay (LETIA)

 

Linearity

The high-value RBP4 serum was two-fold serially diluted with physiological saline, and measured on our CUSAg LETIA platform. Fig.2 showed that the measured RBP4 concentrations were gradually declined along with the serial dilution of blood samples.


Fig.2 Determination of the RBP4 concentration by serial dilution of clinical serum


Accuracy 

Two levels of RBP4 blood samples were analyzed in replicates of three using ouranti-RBP4 monoclonal antibodies on the CUSAg LETIA platform. Data from this study are summarized in the following table, the relative bias was <1%.


Table 1. Analysis of the accuracy of CUSAg RBP4 assay

 

Determined Con.(mg/L)

Mean Con. (mg/L)

Bias (%)

1(15.5 mg/L)

15.6

15.9

15.4

15.6

0.6

2(24.8 mg/L)

24.6

24.8

24.5

24.6

0.8


Precision

Blood samples of control low and control high were repeatedly measured 10 times using self-made reagents with CUSAg RBP4 antibody. The CV% of two are all <1%, as shown in table 2.


Table 2. Analysis of the precision ofCUSAg RBP4 assay

   Control

Mean Con.(mg/L)

   SD        

   CV(%)  

Control low

         30.0

  0.2

    0.7

Control high

          73.2

  0.1

    0.1


Stability

Mouseanti-humanRBP4 monoclonalantibody was prepared into latex reagents, and then the antibodieswereheat-treatedat37°Cfor 14daysand used to detect RBP4  antigens.Table3 revealed the detection performance ofmonoclonalantibodies exposed at 37°C had no significant change compared to that at 4°C. These results showed that the antibody was thermostable.

 

Table 3. Comparison of detection performances of anti-human RBP4 monoclonal antibodies at 37°C and 4°C for 14 days

RBP4 Concentration  (mg/L)

ΔABS

(antibodies stored at 4°C for 14 days)                        

ΔABS

(antibodies treated at 37°Cfor 14 days)

Relative bias(%) 

25

964

957

0.7

50

2129

2243

5.4

100

4426

4556

2.9


Clinical Comparison

Mouseanti-humanRBP4 monoclonalantibodywas evaluated in medium-scale clinical tries with blood samples. Fig.3 shows the comparison of the results of RBP4 measurements determined bythe self-madereagents with CUSAg RBP4antibodiesandthehigh-quality comparison kit.


Fig. 3 Determination of serum RBP4 by self-made reagents using CUSAg RBP4 antibody and the high-quality kit


RBP4 Protein

A certain amount of excellent RBP4 protein (Cat: CSB-DP002A) is also offeredby CUSAg . It can be used as calibrator in immunoassay.

 

Reference

1. Impact of family history on relations between insulin resistance, LDL cholesterol and  carotid IMT in healthy adults. Heart. 2010.

2. Glucose transport and sensing in the maintenance of glucose homeostasis and metabolic harmony.The Journal of Clinical Investigation. 2006,116 (7): 1767–75.

3. Biochemical Basis for Dominant Inheritance, Variable Penetrance, and Maternal Effects  in RBP4 Congenital Eye Disease.Cell. 2015,161 (3): 634–646.


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