Human Cystatin B Quantikine ELISA Kit Summary
Cell Culture Supernates, Serum, EDTA Plasma, Heparin Plasma, Saliva, Urine, Human Milk
|Intra-Assay Precision||Inter-Assay Precision|
The recovery of Cystatin B spiked to three different levels throughout the range of the assay in various matrices was evaluated.
|Sample Type||Average % Recovery||Range %|
|Cell Culture Media (n=4)||102||90-115|
|EDTA Plasma (n=4)||96||86-103|
|Heparin Plasma (n=4)||90||81-97|
Human Cystatin B ELISA Standard Curve
Preparation and Storage
Background: Cystatin B
Cystatin B, also known as Stefin B or liver thiol proteinase inhibitor, is a member of family 1 of the cystatin superfamily. It is a 98 amino acid, intracellular inhibitor regulating the activities of cysteine proteases of the papain family such as Cathepsins B, H and L.
Assay ProcedureRefer to the product
- Prepare all reagents, standard dilutions, and samples as directed in the product insert.
- Remove excess microplate strips from the plate frame, return them to the foil pouch containing the desiccant pack, and reseal.
- Add 100 µL of Assay Diluent to each well.
- Add 50 µL of Standard, control, or sample to each well. Cover with a plate sealer, and incubate at room temperature for 2 hours.
- Aspirate each well and wash, repeating the process 3 times for a total of 4 washes.
- Add 200 µL of Conjugate to each well. Cover with a new plate sealer, and incubate at room temperature for 2 hours.
- Aspirate and wash 4 times.
- Add 200 µL Substrate Solution to each well. Incubate at room temperature for 30 minutes. PROTECT FROM LIGHT.
- Add 50 µL of Stop Solution to each well. Read at 450 nm within 30 minutes. Set wavelength correction to 540 nm or 570 nm.
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