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Q: Do you offer standards or controls in every ELISA kit? We offer positive and negative controls in qualitative ELISA kits, while we offer standards which can work as quality control in quantitative ELISA kits. Kit standards and the entire reagent system are matched. The customer can draw a good standard curve through the experiment, which proves that the whole system including the standards works well, so the standards can play a role of quality control.
We suggest you run duplicates for the standards. Q: What kind of sample types have been validated with your ELISA kit? Can I use your ELISA kit for the sample type which is not mentioned in the manual? Different ELISA kit has been validated in different sample types varying from serum, plasma, body fluid like urine, cell culture supernatant or cell/tissue lysate. Please refer to the kit protocol for sample types. Theoretically, if the concentration of the target is within the detection range, it can be detected out.
We would like to suggest you do a pretest with 24T trial kit first if you want to test the sample type which is not mentioned in the manual. Q: What kind of species is your ELISA kit specific to?
Can I use your ELISA kit for the species which is not mentioned in the manual? Different ELISA kit is designed for different species varying from human, rat, horse, rabbit, mouse, sheep, pig, bovine, dog, chicken, fish, monkey, plant, etc. Please refer to each protocol for species instruction. For different species, the antibody's specificity is different, and there is matrix interference. We can recommend the right kit if you have difficulty to find the kit of species you need. Q: How much sample volume is needed?
For a 96T double antibody ELISA kit, the sample volume needed for testing one sample is 100μL, but considering the loss during the assay, you can prepare 120 μL. We suggest you run duplicated wells, so in this way it takes about 250 μL for testing one sample. For a 96T competitive ELISA kit, the sample volume needed for testing one sample is 50μL, but considering the loss during the assay, you can prepare 70μL. We suggest you run duplicated wells, so in this way it takes about 150μL for testing one sample. Q: What is TMB? Why should I use dual-spectrum to test my sample using TMB system? TMB(3,3',5,5'-Tetramethylbenzidine) is chromogenic substrate used in ELISA.
TMB can act as a hydrogen donor for the reduction of hydrogen peroxide to water by peroxidase enzymes such as horseradish peroxidase. The reaction will be terminated and the color developed in the wells will turn from blue to yellow upon addition of the stop solution. The absorbance value should be read at 450 nm.
We recommend you use dual-spectrum to test your sample to avoid the error caused by interference and scratch in the bottom. Q: Why is yellow or light yellow color developed in the wells of the whole plate when stop solution is added?
Incorrect reagents added. You should check the components and Lot No.
Of reagents to ensure correct reagents of the kit. Insufficient plate washing. You should be sure all wells are filled with buffer with the same volume during every wash step. After final wash, blot plate forcefully on paper towel to remove residual buffer. Contamination of the pipette tips and chromogenic agent container with the enzyme conjugate or contamination of blank wells with positive control. You should change pipette tips between reagents and use separate reservoirs for each reagent.
Use pipette during operation. Substrate exposed to light or contaminated prior to use.
You should keep substrate in the dark until ready to dispense into wells. Concentration of detection antibody or avidin-HRP is too high. You should check calculations or try again after further dilution. Incubation time or color developing time is too long.