A: On each 96 well plate, we recommend running an 8-point standard curve with duplicate wells. With the remaining 80 wells, 40 samples can be tested in duplicate.
A: There are two solutions for insufficient sample volume.
1. Dilute the samples with the provided sample diluent buffer into 100ul.
2. If 50ul of sample is added into a well, add 50ul of standard solution.
A: For tissue homogenates, endogenous biotin should be considered.
Endogenous biotin in tissue homogenates might introduce false positive signal.
Please run the test as described below to confirm if there is Endogenous biotin in the tissue lysate samples using the ELISA kit.
• Add tissue homogenates into the wells and then add ABC and TMB without adding any biotinylated detection antibody to see if any signal will be observed.
• If no signal is produced, then you can work on the tissue sample by using the kit.
A: You can dilute the 10X TBS stock to 1XTBS and test the pH value of the 1xTBS.
The 1XTBS can be used if the pH value falls in the range of 7.2-7.6.
A: Our kits would work normally with samples from Wistar Rat.
A: You can prepare a concentrated wash buffer (25X) which can be stored at 4°C for 1-3 months.
A: Yes, the TMB color developing agent contain H2O2.
A: None of the components in our ELISA kits are produced in humans or primates.
A: The volume of the control is around 100pg. If unopened, the shelf life of the control is the same as the whole kit - "Store at 4˚C for 6 months, at -20˚C for 12 months." The reconstituted control can be stored at -20˚C for 2 days.
A: Using the sample diluent buffer for antibody dilution instead of the antibody diluent buffer will negatively affect the test result to some extent. The values of both standard and sample might be lower than the normal values.
A: Using RIPA lysis system will not have any effect on the ELISA.
A: RIPA lysis buffer is the preferred buffer for lysate preparation because it lyses tissues more thoroughly than PBS.
A: Yes, baculovirus is used during the production of EK0309 as a transfection factor.
A: Yes, sf21 is an insect cell expression system.
A: EK0309 can measure both pro and mature BDNF. However, EK0309 cannot differentiate between pro BDNF and mature BDNF.
A: Yes, it is ok to use common PBS recipes. We have tried many types of buffers with common constituents, and no significant difference was observed whatsoever.
A: We detected the kit in our lab and got our values on the datasheet before delivery, but protein activity will decrease as time goes on, so you may get lower O.D. values than ours. However, it should still be in a reasonable range and the standards can be used to calculate sample values. Good linearity of curve is more important than the actual numerical value.
A: It means Avidin-Biotin-Peroxidase Complex(AR1103) and you could find it in Kit Components.
A: We test serum and plasma routinely, and there is very little difference between serum, plasma and whole blood. The whole blood also contains proteins we need to test. The kit can be used to test whole blood in theory.
A: 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.
A: The acid may affect the binding of antigen to antibody, it is not recommended to use. That being said, it is unlikely to affect the reaction if the solution remains an overall neutral environment.
A: The NIBSC/WHO 1st International Standard is evaluated, we always test our kits before delivery, and customers can find the test result on the protocol. If the customer needs technical support from us to analyze their data, please contact us and we ask that you provide us the following information: the lot# and production date, and when did the customer detect the kit.
A: Double check the protocol for plate washing. It should be examined on three aspects: 1) Did you make the washing buffer correctly? 2) How long did you let the buffer stay in wells? 3) What was the volume of the washing buffer added to each well? In addition, pay attention when adding sample to avoid contamination. And we suggest testing the standards again. Values of standards at low concentration are more affected than that at higher concentration, so customer can still get expected values at high concentration even if there is an error. If problems persist, please contact technical support with the catalog and lot number of your product.
A: The well depth is 300ul, and the max capacity is 350ul. The height of the well is 1.1 cm, and the internal size is 1 cm.
A: The kit does not contain protein produced by viral system, the express system of it is sf21.
A: Our Elisa kits do not come with wash solution, but we have included information about how to make washing buffer on the datasheet. Please refer to the "Material Required But Not Provided" section. We also offer washing buffer for sale separately (Phosphate Buffered Saline Powder SKU: AR0030).
A: This information can be found for each kit under the "Properties" section, and you can find the immunogen sequence information in the "Overview" section.
A: The kit can be used within a month sequentially if it's opened and stored at 4 degree. There is no need to use sealants, the plate can be packaged with aluminum foil bag, and for other reagents keep bottle tightly closed.
A: Our Elisa kits contain 0.02% Sodium Azide. All of the components except TMB colour developing reagent and stop solution contains 0.02% Sodium azide as the preservative.
A: We can provide a recombinant protein as a control. It costs $50 per control and takes 2 weeks to manufacture. We cannot provide a positive or negative control in serum.
A: The knockout (KO) model may contain truncated forms of the target protein which can be detected by the ELISA assay.
A: The plate should soak in 0.3mL PBS or TBS wash buffer for at least 1-2 minutes in an automated wash station.
A: While it is not recommended to use Tween in your wash buffer, small amounts (<0.1% concentration) may decrease background due to insufficient blocking.
A: Our plates are made with the Corning costar plates similar to the DNA-BIND 96 -well plates.
A: 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.
A: 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).
A: Please contact us for negative control suggestions. You can also check expression databases such as genecards, uniprot etc. Due to logistic reasons, we do not sell serum or lysates that we use internally for positive or negative control.
A: You can find Boster's troubleshoot guides under tech support tab. Please contact us for further assistance on troubleshooting your experiment.
A: We have reviewed literature and have gathered this information for most of our ELISA kits. You can find this information on the product page or contact us if you cannot find it. However this information is only suggestive and cannot replace pilot studies in determining the optimal sample dilution ratio.
A: Yes the plate is separable if the kit says that there are removable strips. There are 12 strips of 8 wells each, all removable from the plate. The amount of samples you can run depend on a few factors. In the most common ELISA set up, you will use two strips for standards, and 10 strips for samples using duplicates, which let you run up to 40 samples per kit. Contact us if you have questions regarding other situations.
A: Since different sample types require different reagents, we do not include them in the ELISA kit. However we do have each reagent mentioned in the file below available at very reasonable prices. Be sure to check them out. For sample preparation protocols please download the file below: https://www.bosterbio.com/media/pdf/Boster_Sample_Preparation_Protocols.pdf
A: In theory the ELISA kit will work for all sample types if the target protein is present at a level that falls within the linear range of the ELISA kit detection range. We guarantee the kit to work on the sample types that we have tested. If you want dilution ratio suggestions on these sample types please contact our technical support. For sample types that we have not tested for, we suggest you run pilot experiments to decide the optimal sample dilution.
A: In general, unless otherwise specified, the ELISA kit is pan-specific, meaning that it will react with all different forms of the target protein if they share the majority of the target protein's sequence. The capture and detection antibodies are reactive to the entire sequence of the standard protein. You can find the sequence information of the standard protein in the "immunogen" section. For more information about the specificity of the kit for your particular experiment, please contact our technical support.
A: If the kit is reactive to another commonly used species (human, mouse, and/or rat), we would have listed it as a separate product. If you want to check cross-reactivity to a species that is not included in the 3 species listed above, please contact our technical support for more information. As a rule of thumb, if the sequences are 90%+ identical, there is a high chance of cross-reactivity for your species of interest.
