Enzyme Linked Immunosorbent Assay (ELISA) Operation Procedure (Sandwich Method) - Elabscience

(www.elabscienc...) This is a video about sandwich ELISA kit.
Sandwich ELISA (or sandwich immunoassay) is the most commonly used elisa method. This format requires two antibodies/antigens specific for different epitopes of the antigen/antibodies. These two antibodies/antigens are normally referred to as matched antibody/antigen pairs. One of the antibodies/antigen is coated on the surface of the micro plate and used as the capture antibody/antigen to immobilize the tested analyte. The other antibody/antigen is conjugated with biotin or other markers to facilitate the detection of the analyte.
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[@anandsr5931]

How can i convert ng/ml to iu/L. Cam you please mention specific activity value?

[@Elabscience_Bio]

Hi anandsr5931, I'm sorry to tell that there is no direct conversion between ng/mL and IU/L. The unit can only be converted when we know how much biological activity in 1ng products by assays. But we have no condition to detect the activity of the standard at present.

[@aakifinayat8234]

do we need to dilute and make different concentrations of our sample too?? just like you did it for the standard?

[@Elabscience_Bio]

Due to the different expression levels of different targets in different sample types, it is recommended to refer to relevant literature and combine the detection range of the kit to conduct pre-experiments to determine the appropriate dilution multiple.

[@aakifinayat8234]

how many samples did you tested? and you tested each sample in duplicates? i have 2 samples but I am confused how to prepare my plate? if someone can help me out

[@Elabscience_Bio]

The sample type of our internal verification is normal sample, because there are differences in sample type and sample quantity. In general, such as serum/plasma samples, the number of generally verified is about 16. Sample testing is generally performed with 2 replicates. For the detection of only 2 samples in total, you can choose a small ELISA kit, such as 48T kit, the standard curve and the sample are done with 2 replicates, and the specific operation can be seen in the instruction manual.

[@rajputananigam9401]

I want to.ask you are using dupicate wells for patient sample and same for standard?

[@Elabscience_Bio]

In general, for the accuracy of the test results, it is recommended that both the standard and the sample be dupicate wells.

[@rajputananigam9401]

Can we use single standard only

[@Elabscience_Bio]

If there is no problem with your operation, and the number of samples is relatively large, you can choose standard to do a single well.

[@walternguyen1304]

Sorry, can I ask a question: Why did you mix, vortex, and centrifuge the mixture containing 500ul Reference Standard & Sample Dilute from a 1ml vial (that you stand for 1-2mins) and 500ul from one of 7 tubes maintaining the same solution? Whether these solution has the same sources (from a bottle tagged name with Reference Standard & Sample Dilute)

[@Elabscience_Bio]

Yes, we use the same bottle of standard & sample diluent that comes with the kit.

[@-_BillySaputraS]

does elisa kit comes with its reagent and its buffer?

[@Elabscience_Bio]

Our pre-coated ELISA kits contain a complete set of reagents and detailed operating manual for ELISA experiments, which do not require additional reagents and buffers to be purchased.

[@sanjaykumar-ew6mb]

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