|+1 404 474 4654|
|+1 888 205 9894 (TF)|
bEP ELISA Kit
|2 references available|
|Certificates||ISO 9001:2008, ISO 13485:2003|
|Price||750.32 $ Plus shipping costs $45.00|
|Availability||Will be delivered in 9 to 11 Business Days|
|Method type||Competition ELISA|
|Cross-Reactivity (Details)||No significant cross-reactivity or interference between mouse bEP and analogues was observed. Note: Limited by current skills and knowledge, it is impossible for us to complete the cross- reactivity detection between mouse bEP and all the analogues, therefore, cross reaction may still exist.|
|Sample Type||Serum, Plasma, Biological Fluids|
|Plate||Pre-coated 12 × 8 strip|
|Specificity||This assay has high sensitivity and excellent specificity for detection of mouse bEP.|
|Sensitivity||The sensitivity of this assay, or Lower Limit of Detection (LLD) was defined as the lowest protein concentration that could be differentiated from zero. It was determined by subtracting two standard deviations to the mean optical density value of twenty zero standard replicates and calculating the corresponding concentration.|
|Minimum Detection Limit||4.56pg/mL|
|Detection Range||12.35 -1,000pg/mL|
Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level mouse bEP were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, middle and high level mouse bEP were tested on 3 different plates, 8 replicates in each plate.
CV(%) = SD/meanX100
Intra-Assay: CV < 10%
Inter-Assay: CV < 12%
The standard curve concentrations used for the ELISA’s were 1,000pg/mL, 333.33pg/mL, 111.11pg/mL, 37.04pg/mL, 12.35pg/mL.The stability of ELISA kit is determined by the loss rate of activity. The loss rate of this kit is less than 5% within the expiration date under appropriate storage condition. The loss rate was determined by accelerated thermal degradation test. Keep the kit at 37°C for 3 days, and compare O.D.values of the kit kept at 37°C with that of at recommended temperature. (referring from China Biological Products Standard, which was calculated by the Arrhenius equation. For ELISA kit, 1 day storage at 37°C can be considered as 2 months at 4°C, which means 3 days at 37°C equaling 6 months at 4°C).
|Principle||The kit is a competitive inhibition enzyme immunoassay technique for the in vitro quantitative measurement of bEP in mouse serum, plasma and other biological fluids.|
This assay employs the competitive inhibition enzyme immunoassay technique. A monoclonal antibody specific for mouse bEP has been pre-coated onto a microplate. A competitive inhibition reaction is launched between biotin labeled mouse bEP and unlabeled mouse bEP (Standards or samples) with the pre-coated antibody specific for mouse bEP. After incubation the unbound conjugate is washed off. Next, avidin conjugated to Horseradish Peroxidase (HRP) is added to each microplate well and incubated. The amount of bound HRP conjugate is reverse proportional to the concentration of bEP in the sample. After addition of the substrate solution, the intensity of color developed is reverse proportional to the concentration of bEP in the sample.
Serum: Allow samples to clot for two hours at room temperature or overnight at 4°C before centrifugation for 20 minutes at approximately 1000×g. Assay immediately or store samples in aliquot at -20°C or -80°C. Avoid repeated freeze/thaw cycles.
Plasma: Collect plasma using EDTA or heparin as an anticoagulant. Centrifuge samples for 15 minutes at 1000×g within 30 minutes of collection. Remove plasma and assay immediately or store samples in aliquot at -20°C or -80°C. Avoid repeated freeze/thaw cycles.
Other biological fluids: Centrifuge samples for 20 minutes at 1000×g. Remove particulates and assay immediately or store samples in aliquot at -20°C or -80°C. Avoid repeated freeze/thaw cycles.
|Calculation of Results||This assay employs the competitive inhibition enzyme immunoassay technique, so there is an inverse correlation between bEP concentration in the sample and the assay signal intensity. Average the duplicate readings for each standard, control, and samples. Create a standard curve on log-log or semi-log graph paper, with the log of bEP concentration on the y-axis and absorbance on the x-axis. Draw the best fit straight line through the standard points and it can be determined by regression analysis. Using some plot software, for instance, curve expert 1.30, is also recommended. If samples have been diluted, the concentration read from the standard curve must be multiplied by the dilution factor.In order to make the calculation easier, we plot the O.D. value of the standard (X-axis) against the log of concentration of the standard (Y-axis), although concentration is the independent variable and O.D. value is the dependent variable. The O.D. values of the standard curve may vary according to the conditions of assay performance (e.g. operator, pipetting technique, washing technique or temperature effects). Typical standard curve below is provided for reference only.|
Limited by the current condition and scientific technology, we cant completely conduct the comprehensive identification and analysis on the raw material provided by suppliers. So there might be some qualitative and technical risks to use the kit.
The final experimental results will be closely related to validity of the products, operation skills of the end users and the experimental environments. Please make sure that sufficient samples are available.
Kits from different batches may be a little different in detection range, sensitivity and color developing time.
Do not mix or substitute reagents from one kit lot to another. Use only the reagents supplied by manufacturer.
Protect all reagents from strong light during storage and incubation. All the bottle caps of reagents should be covered tightly to prevent the evaporation and contamination of microorganism.
There may be some foggy substance in the wells when the plate is opened at the first time. It will not have any effect on the final assay results. Do not remove microtiter plate from the storage bag until needed.
Wrong operations during the reagents preparation and loading, as well as incorrect parameter setting for the plate reader may lead to incorrect results. A microplate plate reader with a bandwidth of 10nm or less and an optical density range of 0-3 O.D. or greater at 450 ± 10nm wavelength is acceptable for use in absorbance measurement. Please read the instruction carefully and adjust the instrument prior to the experiment.
Even the same operator might get different results in two separate experiments. In order to get better reproducible results, the operation of every step in the assay should be controlled. Furthermore, a preliminary experiment before assay for each batch is recommended.
Each kit has been strictly passed Q.C test. However, results from end users might be inconsistent with our in-house data due to some unexpected transportation conditions or different lab equipments. Intra-assay variance among kits from different batches might arise from above factors, too.
Kits from different manufacturers for the same item might produce different results, since we havent compared our products with other manufacturers.
|Handling Advice||To minimize extra influence on the performance, operation procedures and lab conditions, especially room temperature, air humidity, incubator temperature should be strictly controlled. It is also strongly suggested that the whole assay is performed by the same operator from the beginning to the end.|
|Precaution of Use||The Stop Solution suggested for use with this kit is an acid solution. Wear eye, hand, face, and clothing protection when using this material.|
|Material not included||
|Expiry Date||six months|
|Restrictions||For Research Use only|
Boehncke, Hardt, Schadendorf et al.: "Endogenous μ-opioid peptides modulate immune response towards malignant melanoma." in: Experimental dermatology, Vol. 20, Issue 1, pp. 24-8, 2010 (PubMed).
Li, Liang, Li et al.: "Physiology and cell biology of acupuncture observed in calcium signaling activated by acoustic shear wave." in: Pflügers Archiv : European journal of physiology, Vol. 462, Issue 4, pp. 587-97, 2011 (PubMed).