What are the characteristics of the anti trypsin assay kit

The anti trypsin assay kit (taking the human alpha 1 anti trypsin ELISA assay kit as an example) has the following characteristics:

1. Detection principle and sensitivity

Double antibody sandwich method:

Using enzyme-linked immunosorbent assay (ELISA) technology, the target protein in the sample is captured by pre coated alpha 1 anti trypsin (alpha 1-AT/AAT) antibodies, and then HRP labeled detection antibodies are added to form a sandwich structure, which is finally quantitatively analyzed by substrate TMB colorimetric assay.

High sensitivity and specificity:

Sensitivity: The minimum detection concentration is less than 0.1U/L, which can accurately detect low abundance samples.

. Specificity: Does not cross react with structurally similar substances (such as other serum proteins) to ensure accuracy of results.

Linear range: The correlation coefficient R value between standard linear regression and expected concentration is ≥ 0.9900, showing a good linear relationship in the range of 1-20 nmol (colorimetric method) or 0.1-2 nmol (fluorescence method).

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II. Sample Types and Processing

Diversity Compatibility:

Suitable for various biological samples such as serum, plasma, tissue homogenate, cell culture supernatant, etc., to meet different experimental needs.

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Serum/plasma: Centrifuge to remove sediment and avoid repeated freezing and thawing.

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Tissue homogenate: Grind with PBS buffer and centrifuge to obtain the supernatant. If necessary, add protease inhibitors.

. Cell culture: Centrifuge the supernatant directly for detection.

Preprocessing requirements:

Hemolytic samples should not be tested (hemoglobin interference results).

. Turbidity samples need to be clarified by centrifugation before detection. After the frozen sample is dissolved, it should be thoroughly mixed to avoid local concentration of proteins.

III. Convenience and Efficiency

Standardization Process:

The reagent kit provides detailed instructions, including parameters such as standard dilution, sample dosage, incubation time, etc., with clear operating steps.

. The colorimetric/fluorescence method supports high-throughput detection, and the 96 well plate design can simultaneously process a large number of samples.

Rapid detection:

Color development method: The total reaction time is about 1.5-2 hours (including incubation and color development steps).

. Fluorescence method: Faster detection (completed within 30 minutes) is achieved by changing the fluorescence signal (such as λ ex/em=485/530nm).

IV. Accuracy and Reliability of Results

Repeatability Validation:

In Plate Coefficient of Variation (CV)

; 10%, inter board CV<; 15% to ensure the stability of experimental results. The standard provides 6 concentration gradients, supports drawing standard curves, and calculates sample concentrations.

Anti interference ability:

The kit contains protease inhibitors to reduce the interference of endogenous enzymes in the sample on the results.

. The washing buffer effectively removes unbound substances and reduces background noise.

Fifth, Storage and Stability

Long term Storage:

Unopened reagent kits need to be stored at 2-8 ℃ in the dark, with a shelf life of 6 months (some products can last up to 24 months).

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After opening, it should be sealed and stored to avoid repeated freezing and thawing, which may cause the reagent to fail.

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Integrity of supporting reagents:

Includes a complete set of components such as microporous enzyme-linked immunosorbent assay plates, standards, detection antibodies, substrates, stop solutions, etc., without the need for additional reagents.

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VI. Application Scenarios and Limitations

Research and Clinical Auxiliary Diagnosis:

Used for quantitative analysis of α 1 antitrypsin deficiency, liver disease, inflammatory response, and other research.

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It should be comprehensively judged in conjunction with other clinical indicators and cannot be used as a diagnostic basis alone.

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Limitations:

Only for in vitro research use and cannot be directly used for clinical diagnosis.

. High requirements for the operating environment, such as avoiding direct sunlight and controlling the incubation temperature.