Sample
Serum, plasma, cellculture supernatant and other biological samples
Intended Use
In vitro quantitative determination of sIgA concentrations in serum, plasma, cellculture supernatant and other biological samples.
Contents of Kit
The sealed kit can be stored at 2-8 °C. The storage condition for opened kit is specified in the table below:
1. ELISA Microplate(Dismountable), 8×12. Put the rest strips into a sealed foilbag with the desiccant. Stored for 1 month at 2-8°C. Stored for 6 month at-20°C.
2. Lyophilized Standard, 2vial. Put the rest standards into a desiccant bag. Stored for 1 month at 2-8°C. Stored for 6 month at -20°C.
3. Biotin-labeled Antibody(Concentrated, 100×), 120μl. 2-8°C (Avoid Direct Light)
4. HRP-Streptavidin Conjugate(SABC, 100×), 120μl. 2-8°C (Avoid Direct Light)
5. TMB Substrate, 10ml. 2-8°C (Avoid Direct Light)
6. Sample Dilution Buffer, 20ml. 2-8°C
7. Antibody Dilution Buffer, 10ml. 2-8°C
8. SABC Dilution Buffer, 10ml. 2-8°C
9. Stop Solution, 10ml. 2-8°C
10. Wash Buffer(25×), 30ml. 2-8°C
11. Plate Sealer, 5 pieces
12. Product Description, 1 copy
Note: The liquid reagent bottle contains slightly more reagent than indicated on the label. Please use pipette accurately measure and do proportional dilution.
Storage
2-8°C (for sealed box), please do not freeze! See kit label for expiry date
Performance Characteristics
Perform the stability test for the sealed kit at 37°C and 2-8°C and get relevant data.
Precision
Intra-assay Precision: samples with low, medium and high concentration are tested 20 times on same plate.
Inter-assay Precision: samples with low, medium and high concentration are tested 20 times on three different plates.
Detection Range
0.313-20ng/ml
Standard Curve
This product has been tested by Quality Control Department and meets performance specifications mentioned in the manual. (The humidity in the laboratory is 20%-60%, and the temperature is 18°C - 25°C. TMB was balanced to 37°C before color development, and incubated at 37°C for 15 minutes in the darkafter adding the enzyme label plate holes.)
The following assay data are provided for reference, since experimental environment and operation are different. The establishment of standard curve depends on your own assay.
Citations
Publication ()
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Effect of pasteurisation and high-pressure processing on selected bioactive components in human milk – An experimental study
Miroslava Jandová, Michaela Fišerová, Pavla Paterová, Pavel Měřička, Jan Malý, Marian Kacerovský, Eliška Kovaříková, Jan Strohalm, Kateřina Demnerová, Jana Kadavá, Hana Sýkorová, Radomír Hyšpler, Alena Tichá, Dana Čížková, Milan Houška, Aleš Bezrouk
Applications: ELISA
Reactive species: Human
"Abstract: High-pressure processing (HPP) represents a promising alternative to conventional Holder pasteurisation (HoP) used by human milk banks worldwide. The objective of this study was to identify whether the HPP would achieve the same or better retention of the content of selected analytes than the HoP. Samples collected from 15 breast milk donors were processed in four ways: i) no treatment; ii) HoP; iii) HPP in cycles (350 MPa, 4 cycles); iv) continuous HPP (350 MPa, 20 min). The content of secretory immunoglobulin A (sIgA), lactoferrin and lysozyme was determined using commercially available ELISA kits, and the lipase activity was assessed using an A-lipase activity assay kit. Data were compared statistically using paired t-tests. HoP significantly reduced the content of lysozyme and lactoferrin as well as lipase activity (P < 0.001). Cycled HPP significantly decreased lipase activity (P = 0.002), while continuous HPP led to a significant decrease in lysozyme content (P = 0.001) and lipase activity (P = 0.014). Cycled HPP showed high retention of pretreatment levels of lysozyme – median 99 (88; 99%), lactoferrin – 84 (66; 105%), and sIgA content – 83 (28; 117%). Among the studied treatment regimens, the best preservation of initial levels of bioactive components was achieved using HPP at 350 MPa in cycles."
Article snippet: sIgA content was determined using an sIgA ELISA kit (CD Creative Diagnostics, USA) following the manufacturer's instructions. The test sensitivity was < 1.875 ng·mL–1; the detection range was 3.125-200 ng·mL–1. 
Figure 1. Residual secretory immunoglobulin A (sIgA) level (%) after Holder pasteurisation (HoP), cycled highpressure processing (HPP), and continuous HPP.
COA
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