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).

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A: Boster Bio does not evaluate sample stability. Variations in sample collection, processing, and storage may affect the stabilityof samples. It is recommend to assay sample on priority basis collection when possible, or aliquot into single use volumes and store samples frozen. avoid repeat freeze-thaw cycles with the stored samples to limit protein degradation.

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A: Chelating agents such as EDTA, Heparin and Citrate can irreversibly bind metal ions from the functional domain of Bdnf causing degradation of its protein structure. Bdnf may be denatured as a result and may compromise the assay's measurements. The chilating sites could also be too close to the epitopes needed for detection and block the antigen antibody reaction. We have tested the Bdnf ELISA, treating samples with various anticoagulants and decided that heparin, EDTA or citrate can be used for treatment of blood/plasma samples. Do not use other anticoagulents when collecting samples.

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A: we recommend you this article on ELISA data analysis. bosterbio.com/ELISA-data-analysis-instructions. we also provide a convenient online tool that you can use to analyze ELISA data. bosterbio.com/biology-research-tools/ELISA-data-analysis-online

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A: The enzyme is always active. Avoid contaminating the substrate with enzyme prior to the incubation otherwise it compromises the assay with false positive signal.

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A: do not freeze and thaw whole blood. erythrocytes are fragile and, if frozen and thawed, will undergo hemolysis rendering the samples useless. To keep your blood samples to test Bdnf for a later time, you should let the blood clot in glass tubes and separate the serum to freeze for later analysis.

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A: Unfortunately, Boster Bio has not routinely validated tissue homogenates as a sample type for ELISA kits. This does not mean that ELISA kits are not valid for other sample types than we have tested: it means further investigation is needed. One will need to perform a spike and recovery study to determine if an unvalidated sample type will work with a particular kit. To perform a spike and recovery experiment, one should divide a sample into two aliquots. In one of the aliquots, the user should spike in a known amount of the kit standard. a dilution series is performed comparing the spiked versus the unspiked sample. Generally, samples with expected recovery and linearity between 80-120% are considered acceptable. This method can be used to validate any sample type that has not been assessd by Boster Bio. for a more detailed spike and recovery protocol, please contact technical support.
Note: acceptable ranges should be determined individually by each laboratory. Additionally, technical support can help determine if a buffer component is not compatible with a given ELISA kit. please check the Citations tab on the product webpage for peer-reviewed papers utilizing a wide range of sample types. We also have an innovator's reward program where if the user validates our ELISA kits in applications or samples previously not validated by Boster Bio or other users, and share such information with us by submit a review, we will reward the user's efforts with a free antibody or ELISA kit from our catalog. Biocompare.com will also give $20 Amazon giftcard as an additional reward, if the review is submitted there as well.

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The bubbles would not affect the result of the Mouse BDNF ELISA Kit PicoKine™ (EK0309), Please wash the plate to remove the bubbles as suggested below prior to the assay. Wash the plate 3 times with the 1x wash buffer. a. Discard the liquid in the wells into an appropriate waste receptacle. Then, invert the plate on the benchtop onto a paper towel and tap the plate to gently blot any remaining liquid. It is recommended that the wells are not allowed to completely dry at any time. b. Add 300 µl of the 1x wash buffer to each assay well. (For cleaner background incubate for 60 seconds between each wash). c. Repeat steps a-b 2 additional times.