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The Boster Human CA15-3 Pre-Coated ELISA (Enzyme-Linked Immunosorbent Assay) kit is a solid phase immunoassay specially designed to measure Human CA15-3 with a 96-well strip plate that is pre-coated with antibody specific for CA15-3. The detection antibody is a HRP conjugated antibody specific for CA15-3. The capture antibody is monoclonal antibody from mouse, and the detection antibody is monoclonal antibody from mouse. The kit is analytically validated with ready to use reagents. To measure Human CA15-3, add standards and samples to the wells, then add the HRP conjugated detection antibody. Wash away the unbounded protein and HRP conjugated detection antibody. TMB is substrate to HRP and will be catalyzed to produce a blue color product, which changes into yellow after adding acidic stop solution. Upon addition of the substrate, the density of the yellow product is linearly propotional to Human CA15-3 in the sample. Read the density of the yellow product in each well using a plate reader, and benchmark the sample wells' readings against the standard curve to determine the concentration of Human CA15-3 in the sample. For more information on assay principle, protocols, and troubleshooting tips, see Boster's ELISA Resource Center at https://www.bosterbio.com/elisatechnical-resource-center.
| Product Name | Human CA15-3 ELISA Kit |
|---|---|
| SKU/Catalog Number | EK7030 |
| Description | Sandwich High Sensitivity ELISA kit for Quantitative Detection of Human CA15-3. 96wells/kit, with removable strips. |
| Cite This Product | Human CA15-3 ELISA Kit (Boster Biological Technology, Pleasanton CA, USA, Catalog # EK7030) |
| Validated Species | Human |
| Application | ELISA *Our Boster Guarantee covers the use of this product in the above tested applications. **For positive and negative control design, consult "Tissue specificity" under Protein Target Info. |
| Cross Reactivity | There is no detectable cross-reactivity. |
| Pack Size | 96wells/kit, with removable strips. |
| Sensitivity | 3.8U/ml *Sensitivity, or Lower Limit of Detection (LLD), is the minimum level of target protein the ELISA assay can detect. We measure 20 blank wells and if the O.D. value is 2 standard deviations higher than the blanks' average O.D. the sample can be deemed positive. |
|---|---|
| Assay Range | 10-300U/ml *This assay range is determined using common samples. For samples with low target protein concentrations, users can adjust the standard curve to extend the lower limit of assay range. |
| Sample Type | Serum *The above listed samples are the ones valided with the assay. If you do not see your sample of interest listed, as long as there is enough level of target protein present in the sample, this Picokine? ELISA kit should detect it. **For protocol and tips regarding preparing your sample of interest, please check our ELISA sample preparation guide. |
| Storage | Store the kit at 2°C to 8°C. Keep microwells sealed in a dry bag with desiccants. The reagents are stable until expiration of the kit. Do not expose reagent to heat, sun, or strong light. Avoid multiple freeze-thaw cycles(Shipped with wet ice.) |
| Description | Quantity | Volume | Buffers |
| Anti-Human CA15-3 Pre-coated 96-well strip microplate | 1 | 8 strips of 12 wells | Anti- CA15-3 monoclonal antibody, Polystyrene micro-well plate |
| Human CA15-3 Standards(S1~S4) | 4 | 0.5ml | CA15-3 (from natural protein)(10，50，150, 300) U/ml, 0.02M PBS, 20% new-born calf serum |
| HRP Conjugated anti-Human CA15-3 antibody | 1 | 11ml | HRP Conjugated anti-Human CA15-3 antibody, 0.02M PBS, 20% new-born calf serum, 0.01% azophloxine, from mouse monoclonal antibody |
| Sample diluent | 1 | 11ml | 0.02M PBS, 20% new-born calf serum |
| Controls | 1 | 0.5ml | CA15-3 (from natural protein) U/ml, 0.02M PBS, 20% new-born calf serum |
| 20X Wash Buffer Concentrate | 1 | 15ml | Neutral buffer, contains 0.5% Tween-20 |
| Color Developing Reagent A | 1 | 7ml | Contains 11mmol/L H2O2 |
| Color Developing Reagent B | 1 | 7ml | Contains 2mmol/L TMB |
| Stop Solution | 1 | 7ml | 2mol/L dilute sulphuric acid |
| Plate Sealers | 2 | Piece |
1. Microplate Reader capable of reading absorbance at 450nm.
2. Automated plate washer (optional)
3. Pipettes and pipette tips capable of precisely dispensing 0.5 μl through 1 ml volumes of aqueous solutions. Multichannel pipettes are recommended for large amount of samples.
4. Deionized or distilledwater.
5. 500ml graduated cylinders.
6. Test tubes for dilution.
You can check the tissue specificity below for information on selecting positive and negative control.
*if product is indicated to react with multiple species, protein info is based on the human gene.
Click the images to enlarge.
Human CA15-3 ELISA Kit standard curve
1. For Research Use Only. Not for use in diagnostic procedures.
2. For Laboratory use.
3. Not for Internal or External Use in Humans or Animals.
4. There should be no eating or drinking within work area.
5. Always wear gloves and a protective lab coat.
6. No pipetting should be done by mouth. Handle all specimens and reagents as potentially infectious and biohazardous.
7. Do not add sodium azide to samples as preservative.
8. Do not use external controls containing sodium azide.
9. Use disposable pipette tips to avoid contaminating chromogenic substrate reagent. Discard reagent if it turns blue.
10. Do not pour chromogenic substrate back into container after use.
11. Do not freeze reagents.
12. Do not mix reagents from different kit lot numbers.
13. Keep reagents out of direct sunlight.
14. Handle stop reagent with care, since it is corrosive.
15. Bring all reagents to room temperature.
16. Viscous forensic samples should always be diluted in phosphate buffered saline or distilled water prior to pipetting.
17. Ensure the bag containing the micro-plate strips and desiccant is sealed well, if only a few strips are used.
Serum or plasma should be prepared from a whole blood specimen obtained by acceptable medical techniques. This kit is for use with serum, plasma-EDTA, or plasma-heparin samples.
1. Immediately before testing, prepare samples by diluting using a 1:9 ratio in Sample Diluent provided. Example: add 50ul of sample to 450ul of Sample Diluent and mix well. Discard unused diluted samples. DO NOT DILUTE THE STANDARDS
2. Prepare 1X Wash buffer by adding the contents of the bottle (25 ml, 20X) to 475 ml of distilled or deionized water. Store at room temperature (20-25°C).
Bring all specimens and kit reagents to room temperature (20-25°C) and gently mix.
1. Patient samples should be diluted 10-fold before use. (See Reagent and Sample Preparation above). DO NOT DILUTE THE STANDARDS
2. Secure the desired number of coated wells in the holder. Dispense 25ul of CA15-3 standards, diluted samples, and diluted controls into the appropriate wells.
3. Add 100ul of Antibody-Biotin Conjugate Reagent (blue solution) to all wells. Gently mix for 20-30 seconds at 500-600 rpm.
4. Incubate for 60 minutes at room temperature.
5. Remove liquid from all wells. Wash each well three times with 350ul of 1X wash buffer. After each wash, sharply and firmly tap the upside down plate on absorbance paper or paper towels to remove residual droplets.
6. Dispense 100ul of Enzyme Conjugate (red solution) into each well.
7. Incubate for 60 minutes at room temperature.
8. Remove the contents and wash the plate 3x as described in step 5 above.
9. Dispense 100ul of TMB Solution into each well.
10. Incubate at room temperature for 15 minutes.
11. Stop the reaction by adding 50ul of Stop Solution to each well.
12. Read the absorbance at 450nm (using a reference wavelength of 630nm) with a microtiter plate reader within 15 minutes.
1. Calculate the average absorbance values for each set of reference standards, control, and samples.
1. The standard curve is constructed as follows: against its concentration in U/ml on linear graph paper, with absorbance on the vertical (y) axis and concentration on the horizontal (x)
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