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ELISA kits for testing

Experimental operation guide

Frequently Asked Questions

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Are there any precautions for the pending markers?

The marker to be labeled needs to have a free primary amine or N-terminal, and the sample must not contain Tris, glycerol, proteins with amino groups or reagents.

Can this biotin labeling kit label other proteins besides antibodies?

The principle of the biotin labeling kit is that the ester bond of the activated biotin and the amino group on the labeled material are dehydrated and condensed to form a stable amide bond. In addition, the N-terminal end of the protein can also be labeled. If there is a primary amine on the protein, such as lysine is what can be labeled.

How can I tell if my sample needs to be diluted or if it needs other treatment?

The detection range of the kit is not equivalent to the concentration range of the sample to be tested. It is recommended to estimate the concentration of the sample to be tested from the relevant literature and to determine the actual concentration of the sample through pre-testing before the experiment. For routine sample types, consult ELISA technical support for recommended dilutions and schedule a pre-test. If the concentration of the analyte in the sample is too high or too low, dilute or concentrate the sample appropriately. If the analyte concentration in the sample is well below the minimum detection line of the kit, it is recommended to select a more sensitive kit for the assay.

How does the "repeated freeze-thaw" process mentioned in tissue/cell sample collection work?

Tissue samples: After tissue grinding, place them at -80°C for 1h/liquid nitrogen for 0.5h, and then place them in a 30°C water bath with slight shaking to make them thaw rapidly, and this operation can be repeated 1-2 times. Cell samples: Repeat the above operation of freezing and thawing 2-3 times. In case of membrane proteins, it can be sonicated properly, but the temperature and frequency of sonication need to be controlled. It is recommended to add protease inhibitors to the sample in advance. Routine recommendation: PMSF.

How can hemolysis be avoided as much as possible during collection?

Hemolysis can be caused by a variety of physicochemical factors and toxins. In vitro, hemolysis can be caused by mechanical forceful oscillations, cryogenic freezing, etc. To avoid the effect of hemolysis on test results, please: (1) When collecting blood from a vein, the pumping back pressure should not be too high (2) The amount of blood collected at one time should not be too small (3) The whole blood collected should not be shaken vigorously (4) Centrifugal speed should not be too high (5) The collected whole blood should be processed into serum/plasma as soon as possible, and whole blood should not be frozen and thawed (6) If anesthesia is required for blood collection, anesthetics without hemolytic effect should be used

What species can your kits detect?

Our kits are currently available for human, mouse and rat, with a small number of indicators for specific species such as monkey, rabbit, pig and chicken. For those kits with a species-specific name (e.g. human), the kit is specific to that species. If no species is specified in the instructions, the kit is generic within the regular mammalian species. Please contact technical support to confirm the suitability of the test for other specific species.

How many samples can be tested with the 96T kit? Do I need to do duplicate wells?

For the 96T kit, both standards & samples are not re-well, maximum 88 samples can be detected; standards are re-well, samples are single-well, maximum 80 samples can be detected; standards are single-well, samples are re-well, maximum 44 samples can be detected; standards & samples are re-well, maximum 40 samples can be detected. If 3 wells are done, the number of samples tested will be reduced accordingly. In order to ensure the accuracy of the experimental results, we recommend that both standards and samples do multiple wells, but it is not required that ELISA experiments must do multiple wells / 3 multiple wells, customers can set their own needs according to the multiple well experiment.

Is it possible to reduce the number of wells in the standard curve when there are more samples to be tested?

To ensure the accuracy of the results, it is not recommended to reduce the number of wells of the standard, please ensure the detection of at least 6 different standard concentrations (including blank wells ).

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