96 Well Plate Mouse High Specificity SP-C Sandwich ELISA Kit For
Accurate Quantitative Detection
Cat.No E1260Mo
Standard Curve Range: 0.5ng/ml - 300ng/ml
Sensitivity: 0.26ng/ml
Storage: Store the reagents at 2-8°C. For over 6-month storage refer to
the expiration date keep it at -20°C. Avoid repeated thaw cycles.
If individual reagents are opened it is recommended that the kit be
used within 1 month.
*This product is for research use only, not for use in diagnosis
procedures. It’s highly recommend to read this instruction entirely
before use.
Reagent Provided
Components | Quantity |
Standard Solution (320ng/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 Mouse SP-C Antibody | 1ml x1 |
User Instruction | 1 |
Plate Sealer | 2 pics |
Zipper bag | 1 pic |
Material Required But Not Supplied
- 37°C±0.5°C incubator
- Absorbent paper
- Precision pipettes and disposable pipette tips
- Clean tubes
- Deionized or distilled water
- Microplate reader with 450 ± 10nm wavelength filter
Precision
Intra-Assay Precision (Precision within an assay) Three samples of
known concentration were tested on one plate to assess intra-assay
precision.
Inter-Assay Precision (Precision between assays) Three samples of
known concentration were tested in separate assays to assess
inter-assay precision.
CV(%) = SD/mean x 100
Intra-Assay: CV<8%
Inter-Assay: CV<10%
Intended Use
This sandwich kit is for the accurate quantitative detection of
Mouse Pulmonary Surfatcant-associated Protein C (also known as
SP-C) in serum, plasma, cell culture supernates, cell lysates,
tissue homogenates.
Reagent Preparation
All reagents should be brought to room temperature before use.
Standard Reconstitute the 120μl of the standard (320ng/ml) with 120μl of
standard diluent to generate a 160ng/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 (160ng/ml) 1:2 with standard
diluent to produce 80ng/ml, 40ng/ml, 20ng/ml and 10ng/ml solutions.
Standard diluent serves as the zero standard(0 ng/ml). Any
remaining solution should be frozen at -20°C and used within one
month. Dilution of standard solutions suggested are as follows:
160ng/ml | Standard No.5 | 120μl Original Standard + 120μl Standard Diluent |
80ng/ml | Standard No.4 | 120μl Standard No.5 + 120μl Standard Diluent |
40ng/ml | Standard No.3 | 120μl Standard No.4 + 120μl Standard Diluent |
20ng/ml | Standard No.2 | 120μl Standard No.3 + 120μl Standard Diluent |
10ng/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 |
320ng/ml | 160ng/ml | 80ng/ml | 40ng/ml | 20ng/ml | 10ng/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.