CST3 ELISA 试剂盒

Sample values: Thirteen random human serum samples were tested in the assay.
Values ranged from 532.0 to 905.6 ng/mL with an average of 675 ng/mL.
The normal reference range for serum Cystatin C is 590-910 ng/mL10.
An abnormal serum sample read at 1,384 ng/mL.

Allow the kit reagents to come to room temperature for 30 minutes.
We recommend that all standards and samples be run in duplicate to allow the end user to accurately determine Cystatin C concentrations.
Ensure that all samples have reached room temperature and have been diluted as appropriate prior to running them in the kit.
Assay Buffer Dilute Assay Buffer Concentrate 1:5 by adding one part of the concentrate to four parts of deion- ized water.
Once diluted this is stable at 4 °C for 3 months.
Wash Buffer Dilute Wash Buffer Concentrate 1:20 by adding one part of the concentrate to nineteen parts of deionized water.
Once diluted this is stable at room temperature for 3 months.
Standard Preparation Label seven glass test tubes as #1 through #7.
Briefly spin vial of standard in a microcentrifuge to ensure contents are at bottom of vial.
Pipet 585 μL of Assay Buffer into tube #1 and 250 μL into tubes #2 to #7.
Carefully add 15 μL of the Cystatin C stock solution to tube #1 and vortex completely.
Take 250 μL of the Cystatin C solution in tube #1 and add it to tube #2 and vortex completely.
Repeat the serial dilutions for tubes #3 through #7.
The concentration of Cystatin C in tubes #1 through #7 will be 10, 5, 2.5, 1.25, 0.625, 0.313 and 0.156 ng/mL.
Use all Standards within 2 hours of preparation.

Serum and plasma samples must be diluted ≥ 1:50 with the provided Assay Buffer prior to running in the kit. A dilution of ≥ 1:225 is recommended to detect most samples within the standard curve range. Disease state samples, such as those from tubular kidney disease, may require further dilu- tions up to 1:500 or greater. It is up to the end user to determine the appropriate dilution for their samples. Urine samples must be diluted ≥ 1:4 with the provided Assay Buffer prior to running in the kit. TCM samples should be diluted in TCM and read off a standard curve generated in the same TCM. Any samples with Cystatin C concentrations outside the standard curve range should be diluted further with Assay Buffer or TCM, as appropriate, to obtain readings within the standard curve. Use all samples within 2 hours of dilution.

1. Use the plate layout sheet on the back page of the insert to aid in proper sample and standard identification. Determine the number of wells to be used and return unused wells to foil pouch with desiccant. Seal the ziploc plate bag and store at 4ºC.
   2. Pipet 50 μL of samples or standards into wells in the plate. Pipet 50 μL of Assay Buffer into the zero standard wells.
   3. Incubate at room temperature for 60 minutes. Aspirate the plate and wash each well 4 times with 300 μL wash buffer. Tap the plate dry on clean absorbent towels.
   4. Add 50 μL of the DetectX® Cystatin C Conjugate to each well, using a repeater pipet.
   5. Incubate at room temperature for 30 minutes.
   6. Aspirate the plate and wash each well 4 times with 300 μL wash buffer. Tap the plate dry on clean absorbent towels.
   7. Add 50 μL of the TMB Substrate to each well, using a repeater pipet.
   8. Incubate the plate at room temperature for 30 minutes.
   9. Add 50 μL of the Stop Solution to each well, using a repeater pipet. 10. Read the optical density generated from each well in a plate reader capable of reading at 450 nm. 11. Use the plate reader's built-in 4PLC software capabilities to calculate Cystatin C concentration for each sample.

Average the duplicate OD readings for each standard and sample.
Create a standard curve by reducing the data using computer software capable of generating a four-parameter logistic curve (4PLC) fit.
The sample concentrations should be multiplied by the dilution factor to obtain neat sample values.
Or use the online tool from http://www.myassays.com/arbor-assays-cystatin-c-immunoassay-kit.assay to calculate the data. *The MyAssays logo is a registered trademark of MyAssays Ltd. tyPical data Sample Mean OD Human Cystatin C Conc. (ng/mL) Standard 1 1.595 10 Standard 2 0.810 5 Standard 3 0.399 2.5 Standard 4 0.221 1.25 Standard 5 0.129 0.625 Standard 6 0.082 0.313 Standard 7 0.061 0.156 B0 0.042 0 Sample 1 0.636 3.76 Sample 2 0.094 0.40 Always run your own standard curve for calculation of results.
Do not use this data.