Mouse BDNF ELISA Kit PicoKine™

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Yes, sf21 is an insect cell expression system.

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RIPA lysis buffer is the preferred buffer for lysate preparation because it lyses tissues more thoroughly than PBS.

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EK0309 can measure both pro and mature BDNF. However, EK0309 cannot differentiate between pro BDNF and mature BDNF.

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Using RIPA lysis system will not have any effect on the ELISA.

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We didn't test cell lysate, the kit can work if the concentration of BDNF in cell lysate is in the test range of the kit. The customer can have a try if he wants to detect it. The general preparation of cell lysate is as follows: Collect cells and rinse cells with PBS. Homogenize and lyse cells thoroughly in lysate solution(Cell Protein Extraction Reagent, Catalog# AR0103). Centrifuge cell lysates at approximately 10000 X g for 5 min to remove debris. Aliquots of the cell lysates were removed and assayed.

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The kit does not contain protein produced by viral system, the express system of it is sf21.

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The antibodies used in those ELISA kits are not fully validated on WB. Also, since these antibodies are made from recombinant proteins, they are often less reactive to retrieved antigens in WB or IHC due to the lack of tertiary structure in the retrieved antigens. If you want to test human and mouse BDNF with WB, please use our anti-BDNF antibody (PB9075). Further, you can separately buy our secondary antibodies.

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Yes, baculovirus is used during the production of EK0309 as a transfection factor.

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Try to repeat your experiment by extending your incubation times as follows: samples (150 min), biotinylated antibodies (70-80 min), ABC reagent (45 min) and TMB (30 min).