Human CA15-3 ELISA Kit

Sample

Serum, plasma, and cell culture supernatants

Species Reactivity

Human

Detection Method

sELISA

Intended Use

The Human CA15-3 (Cancer Antigen) ELISA kit is an in vitro enzyme-linked immunosorbent assay for the quantitative measurement of human CA15-3 in serum, plasma, and cell culture supernatants.

Contents of Kit

1. Bring all reagents and samples to room temperature (18 - 25°C) before use.
2. Assay Diluent B (Item E) should be diluted 5-fold with deionized or distilled water before use.
3. Sample dilution: Assay Diluent A (Item D) should be used for dilution of serum and plasma samples. 1× Assay Diluent B (Item E) should be used for dilution of cell culture supernatant samples. The suggested dilution for normal serum/plasma is 30 - 200 fold.
Note: Levels of CA15-3 may vary between different samples. Optimal dilution factors for each sample must be determined by the investigator.
4. Preparation of standard: Briefly spin a vial of Item C. Add 400 μl Assay Diluent A (for serum/plasma samples) or 1× Assay Diluent B (for cell culture supernatants) into Item C vial to prepare a 100 U/ml standard. Dissolve the powder thoroughly by a gentle mix. Add 5 μl CA15-3 standard (100 U/ml) from the vial of Item C, into a tube with 495 μl Assay Diluent A or 1× Assay Diluent B to prepare a 1,000 mU/ml standard solution. Pipette 300 μl Assay Diluent A or 1× Assay Diluent B into each tube. Use the 1,000 mU/ml standard solution to produce a dilution series (shown below). Mix each tube thoroughly before the next transfer. Assay Diluent A or 1× Assay Diluent B serves as the zero standard (0 mU/ml).

5. If the Wash Concentrate (20×) (Item B) contains visible crystals, warm to room temperature and mix gently until dissolved. Dilute 20 ml of Wash Buffer Concentrate into deionized or distilled water to yield 400 ml of 1× Wash Buffer.
6. Briefly spin the Detection Antibody vial (Item F) before use. Add 100 μl of 1× Assay Diluent B (Item E) into the vial to prepare a detection antibody concentrate. Pipette up and down to mix gently (the concentrate can be stored at 4°C for 5 days). The detection antibody concentrate should be diluted 80-fold with 1× Assay Diluent B (Item E) and used in step 5 of Part VI Assay Procedure.
7. Briefly spin the HRP-Streptavidin concentrate vial (Item G) and pipette up and down to mix gently before use, as precipitates may form during storage. HRP-Streptavidin concentrate should be diluted 200-fold with 1× Assay Diluent B (Item E).
For example: Briefly spin the vial (Item G) and pipette up and down to mix gently. Add 50 μl of HRP-Streptavidin concentrate into a tube with 10 ml 1× Assay Diluent B to prepare a 200-fold diluted HRP- Streptavidin solution (don't store the diluted solution for next day use). Mix well.

Storage

The entire kit may be stored at -20°C for up to 1 year from the date of shipment. Avoid repeated freeze-thaw cycles. The kit may be stored at 4°C for up to 6 months. For extended storage, it is recommended to store at -80°C. For prepared reagent storage,
1. CA15-3 Microplate: 1 month at 4°C (Return unused wells to the pouch containing desiccant pack, reseal along entire edge.)
2. Wash Buffer Concentrate (20×): 1 month at 4°C
3. Standard Protein: 1 week at -80°C
4. Detection Antibody CA15-3: 5 days at 4°C
5. HRP-Streptavidin Concentrate: Do not store and reuse.
6. Assay Diluent B: 1 month at 4°C

Precision

Intra-Assay CV%: <10%
Inter-Assay CV%: <12%

Sensitivity

The minimum detectable dose of Human CA15-3 was determined to be 4 mU/ml. Minimum detectable dose is defined as the analyte concentration resulting in an absorbance that is 2 standard deviations higher than that of the blank (diluent buffer).

Standard Curve

These standard curves are for demonstration only. A standard curve must be run with each assay.