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SCCA2 kit

Product Name IMMQUA SCCA2
Generic Name SCCA2 kit
Approval Number 30100EZX00038000
Product Code 326076529
Tests per Kit 48 tests
Storage Conditions 2~8℃
Shelf Life 1 year

Intended Use

  • Measuring SCCA2 in Sera (an Aid for Assessing the Severity of Atopic Dermatitis in Children Aged 15 Years and Younger)

Measurement Principles

This kit utilizes an enzyme-linked immunosorbent assay (ELISA) method to quantify SCCA2 in serum. Samples are added to microtiter plate with 96-wells (solid phase) coated with anti-SCCA antibody, and after washing, a POD-conjugated anti-SCCA2 antibody is added to react, an immune complex is formed. After the unreacted POD-conjugated anti-SCCA2 antibody is removed by washing, a chromogenic substrate is added to perform the enzymatic reaction, and color develops in proportion to the amount of SCCA2 in the samples. The color development is stopped, and the color intensity is measured at 450nm.
SCCA2 concentration in the sample bound to the antibody in the solid phase is reflected in the chromogenic amount associated with the reaction between the POD and the chromogenic substrate, and therefore, SCCA2 concentration is determined by measuring this.

Outline of Measurement Procedure

Preparation of the sample solution (diluted 101-fold serum)
Dispense the prepared standard solution, sample solution, and control sample

(100 µL/well each).
20-30℃ for 16-24 h
[primary reaction]

Wash ( 5 times )
Dispense POD-conjugate solution (100 µL/well)

20-30℃ for 90 minutes
[secondary reaction]

Wash ( 5 times )
Dispense chromogenic reagent ( 100 µL /well )

20-30℃, 30 minutes
[chromogenic reaction]

Dispense stop solution (100 µL/well)
Determination of absorbance, main wavelength (450 nm) and secondary wavelength (550-700 nm).

Kit Components

① Antibody-coated Plate 1 pcs
② Standard 1 mL × 1
③ Specimen Diluent 100 mL×1
④ POD-conjugate (12 mL)×1
⑤ Conjugate Solvent 12 mL×1
⑥ Color Reagent A 6 mL×1
⑦ Color Reagent B 6 mL×1
⑧ Stop Solution 12 mL×1
⑨ Wash Concentrate 100 mL×2
⑩ Control Sample 1 mL×1
Accessories: Plate seal 2 pcs

Standard Curve Example

  • The SCCA2 concentration in the sample is calculated by multiplying the concentration obtained from the standard curve by the dilution factor of the sample.

Reagent Performance

Sensitivity The absorbance of the standard solution is greater at higher SCCA2 concentrations, and the absorbance difference between the 0 ng/mL standard solution and the 0.2 ng/mL standard solution is 1.20 or higher.
Accuracy When a control sample with a known concentration is measured, accuracy is within ±20% of the known SCCA2 concentration is 1.0 ng/mL or higher, and within ±30% of the known SCCA2 concentration is less than 1.0 ng/mL.
Simultaneous Reproducibility When the same sample is measured three times simultaneously, the C.V. value of the measured value is less than 15%.
Assay range 0.3~20 ng/mL
Linearity <±20%
Minimum detection sensitivity 0.3 ng/mL(serum concentration)
Cross-reactivity Does not react with SCCA1
Standard Human recombinant SCCA2
Sample to be assayed Human serum

Clinical Significance

SCCA(squamous cell carcinoma antigen) is a serine protease inhibitor belonging to serpin superfamily, which is a protein with a molecular weight of about 45 kDa produced mainly by epithelial cells.
SCCA contains the isoforms SCCA1(SERPIN B3) and SCCA2(SERPIN B4, which are independent gene products transcribed from distinct loci, albeit with very high-amino-acid homologies of 91%.It is known that interleukin 13 (IL-13), one of the type Th2 cytokines, plays crucial roles in allergic inflammation. SCCA2 was identified as one of the molecules whose expression is elevated when airway epithelial cells are stimulated with IL-4 and IL-13, cytokines that are central in allergic responses. Serum SCCA2 levels in children with atopic dermatitis have been reported to be significantly elevated according to the severity of the disease, even when compared with existing blood tests. In the treatment of atopic dermatitis, it is important to select and use anti-inflammatory drugs appropriately according to their severity. It was suggested that serum SCCA2 could be a useful marker as an indicator of the severity of childhood atopic dermatitis. Clinical performance test results of product in 159 allergic unaffected patients and 176 patients with atopic dermatitis aged from 0 to 15 years are shown below.