Rat 20S-PSM(20S-Proteasome) ELISA Kit
Elabscience's Rat 20S-PSM ELISA Kit
The Elabscience's Rat Rat 20S-PSM ELISA Kit is used in the invitro quantitative determination of Rat Rat 20S-PSM concentrations in biological fluids.
|Detection Range||1.56-100 ng/mL|
|Specificity||This kit recognizes Rat 20S-PSM in samples, and no significant cross-reactivity or interference between Rat 20S-PSM and analogs was observed.|
|Repeatability||Coefficient of variation is less than 10%|
This Elabscience's Rat 20S-PSM ELISA Kit works on the principle of Competitive-ELISA. The microplate of this kit has a pre-coating of Rat 20S-PSM. The Rat 20S-PSM in the sample or standard competes with Rat 20SPSM on the solid phase supporter for sites on the Biotinylated Detection Ab which is specific to Rat 20S-PSM during the reaction.
- Wash the unbound sample or standard and excess conjugate from the microplate.
- Add avidin conjugated with Horseradish Peroxidase (HRP) to each well and incubate.
- Add a TMB substrate solution to each well.
- Terminate the enzyme-substrate reaction by adding a stop solution and spectrophotometrically measure the color change at 450 nm ± 2 nm.
- Determine the concentration of Rat 20S-PSM in the samples by comparing the
OD of the samples to the standard curve.
Kit components and Storage
|Elabscience's Micro ELISA Plate (Dismountable)||8 wells ×12 strips||-20℃, 6 months|
|Reference Standard||2 vials|
|Concentrated Biotinylated Detection Ab (100×)||1 vial, 120 μL|
|Concentrated HRP Conjugate (100×)||1 vial, 120 μL||-20°C(shading light), 6 months|
|Reference Standard & Sample Diluent||1 vial, 20 mL||4°C, 6 months|
|Biotinylated Detection Ab Diluent||1 vial, 14 mL|
|HRP Conjugate Diluent||1 vial, 14 mL|
|Concentrated Wash Buffer (25×)||1 vial, 30 mL|
|Substrate Reagent||1 vial, 10 mL||4°C(shading light)|
|Elabscience's Rat 20S-PSM ELISA Stop Solution||1 vial, 10 mL||4°C|
|Plate Sealer||5 pieces|
|Product Description||1 copy|
|Elabscience's Certificate of Analysis||1 copy|
Note: The bottle caps of all reagents must be tightened after use and stored as per mentioned.
Other supplies required
High-precision transfer pipette, Microplate reader with 450 nm wavelength filter, EP tubes and disposable pipette tips, Incubator, Deionized water, Loading slot for Wash Buffer, and Absorbent paper.
1. While carrying out the experiment please wear lab coats, latex gloves, and eye protection. Please follow the national security protocols of biological laboratories.
2. A newly opened ELISA Plate may have a water-like substance, which is normal and will not have any effect on the experiment or its results.
3. The reconstituted standard, concentrated HRP conjugate working solution, and biotinylated detection Ab working solution should not be reused. The unused undiluted stock solutions should be stored according to the mentioned storage conditions.
4. A filter of 450 nm (±10 nm) and a detector that can detect the wavelength should be installed to the microplate reader. The OD should be within the range of 0-3.5.
5. Don't mix or use components from other lots.
6. Pipette tips should be changed in between adding standards, adding samples, and while adding reagent. And also, separate reservoirs should be used for individual reagents.
Note for sample
1. Samples should be assayed within 7 days when stored at 2-8℃.
2. Predict the concentration prior to assaying. Determine the optimal sample dilutions for their particular experiments if the sample concentration is not within the range of the standard curve.
3. The preliminary experiment should be carried out to verify the validity, if the sample type is not included in the manual.
4. There is a possibility of a deviation due to the introduced chemical substance if lysis buffer is used to prepare cell culture supernatant or tissue homogenates.
5. Due to mismatching with the coated antibody or detection antibody, some recombinant protein may not be detected.
Calculation of results for Rat 20S-PSM ELISA
1) Get the average of the duplicate readings for each standard and samples, and then subtract the average zero standard optical density.
2) Plot a four-parameter logistic curve, with standard concentration and OD values on the x-axis and y-axis, respectively. The concentration calculated from the standard curve must be multiplied by the dilution factor if the samples have been diluted. Re-testing should be carried out with an appropriate dilution if the OD of the sample surpasses the upper limit of the standard curve. The actual concentration is the calculated concentration multiplied by the dilution factor.
1. Add 50 μL standard or sample solution to each well. And add 50 μL Biotinylated Detection Ab immediately to each well. Incubate for 45 min at 37℃
2. Aspirate and wash 3 times
3. Add 100 μL HRP Conjugate to each well and incubate for 30 min at 37℃.
4. Aspirate and wash 5 times.
5. Add 90μL Substrate Reagent. Incubate 15 min at 37℃.
6. Add 50μL Stop Solution. Read at 450nm immediately
7. Calculate the results.
Elabscience is a biotechnology company that provides immunodiagnostic technology. They have products like ELISA Kits like the Rat ELISA kit, Antibodies, Peptides, FCM Antibodies, Recombinant Proteins, CLIA Kits, Biochemical Kits, Labeling Kits, GICA Kits, Chromatographic Media, Cell Line and Cell Culture Products and other Reagents providing a complete platform for R&D and manufacture. And they have in house QC for each and every product and have customers in more than 100 countries all over the world.
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