KL-6 ELISA Assay Kit

$475.00

The KL-6 ELISA Assay Kit is for the quantitative determination of Human Krebs von den Lundgen-6 in serum, plasma and cell culture supernatant. The KL-6 ELISA Assay Kit is intended for research use only and not for use in diagnostic or clinical procedures.

KL-6 ELISA Assay Kit

The KL-6 ELISA Assay Kit is For Research Use Only

Size: 12×8 wells
Sensitivity: 1.12U/ml
Standard Range: 20-320 U/ml
Incubation Time: 70 minutes
Sample Type: Serum, cell culture supernatant and plasma
Sample Size: 40 ul
Alternative Name: Krebs von den Lungen-6


Assay Background

Krebs von den Lungen 6 (KL-6) is a sensitive marker for diagnosing, monitoring, and predicting the prognoses of interstitial lung diseases (ILDs). Particularly, KL-6 is elevated in the serum of patients with active interstitial pneumonia, making it an ideal biomarker for detection and study of the therapy for the disease. KL-6 is a type of transmembrane mucoprotein-2 secreted by proliferating or damaged type II alveolar epithelial cells. It is specific in judging the function of type II alveolar epithelial cells. The expression of KL-6 is significantly increased when the alveolar epithelium sustains injuries. The KL-6 can be secreted into the bloodstream through the damaged alveolar basement membrane.


Sample Preparation and Storage:
Specimens should be clear and non-hemolyzed. Samples should be run at a number of dilutions to ensure accurate quantitation.
1. Extract as soon as possible after specimen collection as per relevant procedure. The samples should be tested as soon as possible after the extraction. Alternately the extracted samples can be kept in -20°C. Avoid repeated freeze-thaw cycles.
2. Serum- Coagulate at room temperature for 10-20 minutes; centrifuge for 20-min at 2000-3000 rpm. Remove the supernatant. If precipitation appears, recentrifuge.
3. Plasma- Use EDTA or citrate plasma as an anticoagulant, mix for 10-20 minutes; centrifuge for 15-min at 2000-3000 rpm. Remove the supernatant carefully. If precipitation appears, recentrifuge.
4. Urine- Collect urine in a sterile container, centrifuge for 20-min at 2000-3000 rpm. Remove the supernatant. If precipitation appears, recentrifuge.
5. Cell Culture Supernatant- Collect sample in a sterile container. Centrifuge for 20-mins at 2000-3000 rpm. Remove the supernatant carefully. When examining the components within the cell, dilute cell suspension with PBS (pH 7.2-7.4), if cell concentration is greater than 1 million/ml. Damage the cells by repeated freeze-thaw cycles to release intracellular components. Centrifuge for 20-min at 2000-3000 rpm. If precipitation appears, centrifuge again.
6. Tissue Samples- Rinse tissues in PBS (pH 7.4) to remove excess blood thoroughly and weigh before homogenization. Mince tissues and homogenize them in PBS (pH7.4) with a glass homogenizer on ice. Thaw at 2-8°C or freeze at -20°C. Centrifuge at 2000-3000 RPM for approximately 20 minutes and collect the supernatant carefully.
Note: Grossly hemolyzed samples are not suitable for use in this assay.


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Additional Information

Assay Principle


The method employs sandwich ELISA technique. Monoclonal antibodies are pre-coated onto microwells. Samples and standards are pipetted into microwells and Human Krebs yon den Lundgen-6, KL-6 present in the sample are bound by the antibodies. Biotin labeled antibody is added and followed by Streptavidin HRP is pipetted and incubated to form a complex. After washing microwells in order to remove any nonspecific binding, the substrate solution (TMB) is added to microwells and color develops proportionally to the amount of Human Krebs yon den Lundgen-6, KL-6 in the sample. Color development is then stopped by addition of stop solution. Absorbance is measured at 450 nm.

Documents

Product Manual