Human Cystatin B Quantikine ELISA Kit Summary
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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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Recombinant human CSTB delivered into mouse using virus vector and assayed from mouse Serum. The mouse serum had low values(both in WT and CSTB KO mice) so signal likely not due to mouse CSTB.
Worked as expected in mouse cell line over expressing CSTB.