0
The Bosterbio OneStep ELISA kit is a solid phase direct ELISA sandwich kit. Instead of adding samples detection antibody and ABC-HRP separately, The OneStep ELISA kit allows the user to add standards, samples and controls to wells in one step, along with the incubation buffer. After a simple washing step, an enzyme conjugate reagent is added into each well. After the excess enzyme conjugate is washed out, the substrate is added into each well. The enzyme catalyzes the substrate yielding a blue color (Amax = 370nm and 652nm) that changes to yellow (Amax = 450nm) upon addition of a sulfuric or phosphoric acid stop solution. The intensity of color developed is directly proportional to the concentration of target protein in the samples. A standard curve is generated relating color intensity to the concentration of target protein.
| Product Name | Human Alpha 1 Antichymotrypsin OneStep ELISA Kit |
|---|---|
| SKU/Catalog Number | EK7018 |
| Description | Human Alpha 1 Antichymotrypsin OneStep ELISA Kit, tested with Serum and Plasma. Format: 96wells/kit, with removable strips. |
| Cite This Product | Human Alpha 1 Antichymotrypsin OneStep ELISA Kit (Boster Biological Technology, Pleasanton CA, USA, Catalog # EK7018) |
| 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 | 6.25 ng/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 | 6.25-400 ng/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 and Plasma *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 |
| Microwell plate coated with anti-ACT Polyclonal Ab | 12x8x1 |
| Alpha 1-Antichymotrypsin Standard: 8 vials (ready to use) | 0.2ml |
| Alpha 1-Antichymotrypsin Controls : 2 vials | 0.2ml |
| Anti-ACT Enzyme Conjugate: 1 vial (ready to use) | 12ml |
| Incubation Buffer: 1 bottle (ready to use) | 12ml |
| Sample Diluent: 3 bottles | 3x22ml |
| TMB Substrate: 1 bottle (ready to use) | 12ml |
| Stop Solution: 1 bottle (ready to use) | 12ml |
| 20X Wash concentrate: 1 bottle | 25ml |
1. Distilled or deionized water
2. Precision pipettes
3. Disposable pipette tips. Multichannel pipettes are recommended in the condition of large amount of samples in the detection.
4. ELISA reader capable of reading absorbance at 450nm
5. Absorbance paper or paper towel
6. Graph paper
You can check the tissue specificity below for information on selecting positive and negative control.
| Gene Name | SERPINA3 |
|---|---|
| Uniprot ID | P01011 |
*if product is indicated to react with multiple species, protein info is based on the human gene.
Click the images to enlarge.
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.
1. Alpha 1-Antichymotrypsin is extracted by the sample diluent out of the stool sample.
2. Typically, specimens may be stored refrigerated at (2-8. C) for 5 days. If storage time exceeds 5 days, store frozen at (-20. C) for up to one month.
3. Avoid multiple freeze-thaw cycles.
4. Prior to assay, frozen samples should be completely thawed and mixed well.
1. Samples: Dilute stool samples 1: 1000 in sample diluent.
2. Wash Concentrate: 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).
Before proceeding with the assay, bring all reagents, serum references and controls to room temperature (20-25°C).
1. Format the microplate wells for each standard, control and patient specimen to be assayed in duplicate. Replace any unused microwell strips back into the aluminum bag, seal and store at 2-8°C.
2. Pipette 10ul of the standards, controls and diluted samples into the assigned well.
3. Add 100ul of incubation buffer into all wells.
4. Cover plate and incubate for 60minutes, at room temperature, with shaking (600rpm)
5. Remove liquid from all wells. Wash wells three times with 300ul of 1X wash buffer The standard curve is constructed as follows:
6. Add 100ul of anti-alpha 1-antichymotrypsin enzyme conjugate solution into all wells.
7. Incubate the plate for 30 minutes, at room temperature, with shaking (600rpm).
8. Remove liquid from all wells. Wash wells three times with 300ul of 1X wash buffer The standard curve is constructed as follows:
9. Add 100ul of TMB substrate solution to all wells
10. Cover and incubate the plate for 15 minutes at room temperature.
11. Add 50ul of stop solution to each well and gently mix for 10 seconds.
12. Read the absorbance on ELISA Reader of each well at 450nm within 15 minutes after adding the stop solution.
The standard curve is constructed as follows:
1. Check ACT standard value on each standard vial. This value might vary from lot to lot. Make sure you check the value on every kit. See example of the standard attached.
2. To construct the standard curve, plot the absorbance for Lysozyme standards (vertical axis) axis) versus ACT standard concentrations (horizontal axis) on a linear graph paper. Draw the best curve through the points.
3. Read the absorbance for controls and each unknown sample from the curve. Record the value for each control or unknown sample.
温馨提示:因厂家更改产品包装、产地或者更换随机附件等没有任何提前通知，且每位咨询者购买情况、提问时间等不同，为此以下回复仅对提问者3天内有效，其他网友仅供参考！若由此给您带来不便请多多谅解，谢谢！
服务热线
0771-3293894
新浪微博
腾讯微博
QQ空间
客服
