High Specificity CA15-3 ELISA Test Kit Canine Carbohydrate Antigen
15-3 ELISA ssay Kit 96 Well Plate
Cat.No E0156Ca
Standard Curve Range: 0.2U/ml - 60U/ml
Sensitivity: 0.096U/ml
Size: 96 wells
* This product is for research use only, not for use in diagnosis
procedures. It’s highly recommend to read this instruction entirely
before use.
Intended Use
This sandwich kit is for the accurate quantitative detection of
Canine Carbohydrate Antigen 15-3 (also known as CA15-3) in serum,
plasma, cell culture supernates, cell lysates, tissue homogenates.
Reagent Provided
Components | Quantity |
Standard Solution (64U/ml) | 0.5ml x1 |
Pre-coated ELISA Plate | 12 * 8 well strips x1 |
Standard Diluent | 3ml x1 |
Streptavidin-HRP | 6ml x1 |
Stop Solution | 6ml x1 |
Substrate Solution A | 6ml x1 |
Substrate Solution B | 6ml x1 |
Wash Buffer Concentrate (25x) | 20ml x1 |
Biotinylated Canine CA15-3 Antibody | 1ml x1 |
User Instruction | 1 |
Plate Sealer | 2 pics |
Zipper bag | 1 pic |
Precautions
- Prior to use, the elisa assay kit and sample should be warmed
naturally to room temperature 30 minutes.
- This instruction must be strictly followed in the experiment.
- Once the desired number of strips has been removed, immediately
reseal the bag to protect the remain from deterioration. Cover all
reagents when not in use.
- Make sure pipetting order and rate of addition from well-to-well
when pipetting reagents.
- Pipette tips and plate sealer in hand should be clean and
disposable to avoid cross-contamination.
- Avoid using the reagents from different batches together.
- Substrate solution B is sensitive to light, don’t expose substrate
solution B to light for a long time.
- Stop solution contains acid. Please wear eye, hand and skin
protection when using this material. Avoid contact of skin or
mucous membranes with kit reagent.
- The elisa assay kit should not be used beyond the expiration date.
Reagent Preparation
All reagents should be brought to room temperature before use.
Standard Reconstitute the 120μl of the standard (64U/ml) with 120μl of
standard diluent to generate a 32U/ml standard stock solution.
Allow the standard to sit for 15 mins with gentle agitation prior
to making dilutions. Prepare duplicate standard points by serially
diluting the standard stock solution (32U/ml) 1:2 with standard
diluent to produce 16U/ml, 8U/ml, 4U/ml and 2U/ml solutions.
Standard diluent serves as the zero standard(0 U/ml). Any remaining
solution should be frozen at -20°C and used within one month.
Dilution of standard solutions suggested are as follows:
32U/ml | Standard No.5 | 120μl Original Standard + 120μl Standard Diluent |
16U/ml | Standard No.4 | 120μl Standard No.5 + 120μl Standard Diluent |
8U/ml | Standard No.3 | 120μl Standard No.4 + 120μl Standard Diluent |
4U/ml | Standard No.2 | 120μl Standard No.3 + 120μl Standard Diluent |
2U/ml | Standard No.1 | 120μl Standard No.2 + 120μl Standard Diluent |
Standard Concentration | Standard No.5 | Standard No.4 | Standard No.3 | Standard No.2 | Standard No.1 |
64U/ml | 32U/ml | 16U/ml | 8U/ml | 4U/ml | 2U/ml |
Wash Buffer Dilute 20ml of Wash Buffer Concentrate 25x into deionized or
distilled water to yield 500 ml of 1x Wash Buffer. If crystals have
formed in the concentrate, mix gently until the crystals have
completely dissolved.
Summary
1. Prepare all reagents, samples and standards.
2. Add sample and ELISA reagent into each well. Incubate for 1 hour
at 37°C.
3. Wash the plate 5 times.
4. Add substrate solution A and B. Incubate for 10 minutes at 37°C.
5. Add stop solution and color develops.
6. Read the OD value within 10 minutes.
Calculation of Result
Construct a standard curve by plotting the average OD for each
standard on the vertical (Y) axis against the concentration on the
horizontal (X) axis and draw a best fit curve through the points on
the graph. These calculations can be best performed with
computer-based curve-fitting software and the best fit line can be
determined by regression analysis.