The Rat Cys-C (Cystatin C) Superset Max DIY ELISA is a high-performance “do-it-yourself” development ELISA kit, designed for the quantitative measurement of Cystatin C. This kit includes all essential components required to optimize & scale up to 5 x 96-well ELISA plates.
SuperSet Max DIY ELISA Kit contains: ✔ Carefully optimized capture and detection antibodies ✔ Purified recombinant protein standard ✔ 5 x 96-well ELISA plates ✔ Streptavidin-HRP Conjugate
With premium reagents and pre-validated components, the SuperSet Max range provides researchers with a reliable, flexible solution for measuring analytes in cell culture supernatants, serum, plasma, and other complex biological samples.
*Available in an Accessory Pack for ELISA Set, Assay Genie product AEES03043.
Microtitre plate reader with 450nm filter (optional 630nm reference)
Microplate washer or wash bottle
10, 50, 100, 200, 1000µl micropipettes with disposable tips
50-300µl multi-channel micropipette with disposable tips
Multichannel micropipette reagent reservoirs
Distilled water
Vortex mixer
Miscellaneous laboratory plastic/glass (sterile if possible)
*Note:The below protocol is a sample protocol. Protocols are specific to each batch/lot. For the correct instructions please follow the protocol included in your kit.
Step
Protocol
1.
Determine wells for diluted standard, blank, and sample. Add 100 μL each dilution of standard, blank, and sample into the appropriate wells (It is recommended that all samples and standards be assayed in duplicate. It is recommended to determine the dilution ratio of samples through preliminary experiments or technical support recommendations). Cover the plate with the sealer provided in the kit. Incubate for 90 min at 37℃. Note: solutions should be added to the bottom of the micro ELISA plate well, avoid touching the inside wall and causing foaming as much as possible.
2.
Decant the liquid from each well, do not wash. Immediately add 100 μL of Biotinylated Detection Ab working solution to each well. Cover the plate with a new sealer. Incubate for 1 hour at 37°C.
3.
Decant the solution from each well, add 350 μL of wash buffer to each well. Soak for 1 min and aspirate or decant the solution from each well and pat it dry against clean absorbent paper. Repeat this wash step 3 times. Note: a microplate washer can be used in this step and other wash steps. Make the tested strips in use immediately after the wash step. Do not allow wells to be dry.
4.
Add 100 μL of HRP Conjugate working solution to each well. Cover the plate with a new sealer. Incubate for 30 min at 37°C.
5.
Decant the solution from each well, repeat the wash process for 5 times as conducted in step 3.
6.
Add 90 μL of Substrate Reagent to each well. Cover the plate with a new sealer. Incubate for about 15 min at 37°C. Protect the plate from light. Note: the reaction time can be shortened or extended according to the actual color change, but not more than 30 min. Preheat the Microplate Reader for about 15 min before OD measurement.
7.
Add 50 μL of Stop Solution to each well. Note: adding the stop solution should be done in the same order as the substrate solution.
8.
Determine the optical density (OD value) of each well at once with a micro-plate reader set to 450 nm.
When carrying out an ELISA assay it is important to prepare your samples in order to achieve the best possible results. Below we have a list of procedures for the preparation of samples for different sample types.
Sample Type
Protocol
Serum:
Allow samples to clot for 1 hour at room temperature or overnight at 2-8℃ before centrifugation for 20 min at 1000×g at 2-8℃. Collect the supernatant to carry out the assay.
Plasma:
Collect plasma using EDTA-Na2 as an anticoagulant. Centrifuge samples for 15 min at 1000×g at 2-8℃ within 30 min of collection. Collect the supernatant to carry out the assay. Recommended reagents for sample preparation: 10×EDTA Anticoagulant
Uniport ID:
P21460
Sample Type:
Serum, plasma
Specificity:
This kit recognizes Rat Cys-C in samples.No significant cross-reactivity or interference between Rat Cys-C and analogues was observed
