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ELISA - IBL

  • Question
    Q.Does this kit cross react to BACE2?
  • Answer
    A.Not, it doesn't.
  • Question
    Q.What is recommended dilution ratio for human serum samples?
  • Answer
    A.The recommended dilution ratio for human serum sample is 2 fold.
  • Question
    Q.What is epitope of the antibodies used in this kit?
  • Answer
    A.The HRP conjugated labeled antibody (Anti-Human APP (R101A4) Mouse IgG Fab’ Affinity Purify) used in this kit detects domain at N-Terminal.
    The epitope of the precoated plate antibody (Anti-Human sAPPα (2B3) Mouse IgG MoAb Affinity Purify at C-Terminal) used in this kit is DAEFRHDSGYEVHHQK.
  • Question
    Q.What type of soluble APP can be measured by this kit?
  • Answer
    A.This kit cross reacts to any type of human soluble APP.
  • Question
    Q.Does it cross react to mouse or rat Aβ (1-40)?
  • Answer
    A.No it doesn't.
  • Question
    Q.Does it cross react to mouse or rat Aβ (1-42)?
  • Answer
    A.No it doesn't.
  • Question
    Q.Can mouse or rat samples be used for this kit?
  • Answer
    A.Wild type mouse or rat samples cannot be used, but APP-transgenic (Tg2576) mouse can be used for the kit.
  • Question
    Q.Is there any recommended buffer solution for cell extraction?
  • Answer
    A.We recommend to use our product #19022 IBLysis.
  • Question
    Q.What is the epitope of the antibodies used in this kit?
  • Answer
    A.The epitope of HRP conjugated labeled antibody (Anti-Human Aβ (N) (82E1) Mouse IgG MoAb Affinity Purify at N-Terminal) used in this kit is DAEFRHDSGYEVHHQK.
    The epitope of the precoated plate antibody (Anti-Human APP-C Rabbit IgG Affinity Purify at C terminal) used in this kit is NGYENPTYKFFEQMQN.
  • Question
    Q.Can serum samples be used for this kit?
  • Answer
    A.No, it cannot.
  • Question
    Q.Can serum samples be used for this kit?
  • Answer
    A.Not recommended to use serum samples. Please use EDTA-plasma samples.
  • Question
    Q.What is recommended anticoagulant?
  • Answer
    A.We use 2Na.
  • Question
    Q.Can VEGF-R2 be measured by this kit?
  • Answer
    A.No, it cannot.
  • Question
    Q.What is recommended dilution ratio for serum samples?
  • Answer
    A.It is 2 to 8 fold.
  • Question
    Q.What is the condition of DPP 4 inhibitor?
  • Answer
    A.Only EDTA-plasma collected using collecting tubes contained DPP4 inhibitor can be used for measuring of GLP-1 active form. BD™ P800 GLP-1, GIP, Glucagon, Ghrelin storing vacuumed collecting tube should be used for collecting blood or the blood should be blocked its activity by DPP4 inhibitor prior it is measured. The amount of DPP4 inhibitor for use is 10μL per 1mL blood or 20μL per 1mL plasma.
  • Question
    Q.What kind of degradative enzyme inhibitor is suitable for use?
  • Answer
    A.We use DPP4 inhibitor.
  • Question
    Q.What is recommended dilution ratio for blood samples?
  • Answer
    A.Please start to dilute from 2 fold.
  • Question
    Q.Can serum samples be used for this kit?
  • Answer
    A.Not recommended to use serum samples.
  • Question
    Q.Can serum sample be used for this kit?
  • Answer
    A.Not recommended to use serum samples. Please refer to the following article for comparison data between serum and plasma. https://www.ncbi.nlm.nih.gov/pubmed/?term=25045139
  • Question
    Q.Can frozen serum sample be used for this kit?
  • Answer
    A.Yes, it can. However, stability test of the samples should be conducted and confirmed by each institute.
  • Question
    Q.What is recommended dilution ratio?
  • Answer
    A.8 fold dilution is acceptable for the measurement, however, we recommend to dilute mouse urine sample more than 16 fold to avoid the data becomes range over.
  • Question
    Q.Which isoform of FGF21 can detect by this kit?
  • Answer
    A.This assay kit react to both full length FGF 21 (1-181) and FGF21 (7-181). Therefore, this kit can measure total FGF21.
  • Question
    Q.How much is the volume of sample required for measurement?
  • Answer
    A.5μL is recommended.
  • Question
    Q.How long does the plate coated biotinylated antibody become stable for use?
  • Answer
    A.We have confirmed that it is stable for a week.
  • Question
    Q.How long does streptavidin plate become stable for use after first opening the pack?
  • Answer
    A.We have confirmed that it is stable at 37℃ for 2 weeks.
  • Question
    Q.How much is the volume of sample required for measurement?
  • Answer
    A.5μL is recommended.
  • Question
    Q.How long does the plate coated biotinylated antibody become stable for use?
  • Answer
    A.We have confirmed that it is stable for a week.
  • Question
    Q.How long does streptavidin plate become stable for use after first opening the pack?
  • Answer
    A.We have confirmed that it is stable at 37℃ for 2 weeks.
  • Question
    Q.How much is the volume of sample required for measurement?
  • Answer
    A.2μL is acceptable for measuring GIP Total using samples collected after meal, however at least 5μL is required for measuring it using samples collected in fasting.
  • Question
    Q.How long does the plate coated biotinylated antibody become stable for use?
  • Answer
    A.We have confirmed that it is stable for a week.
  • Question
    Q.How long does streptavidin plate become stable for use after first opening the pack?
  • Answer
    A.We have confirmed that it is stable at 37℃ for 2 weeks.
  • Question
    Q.How much is the volume of sample required for measurement?
  • Answer
    A.2μL is acceptable for measuring GIP Total using samples collected after meal, however at least 5μL is required for measuring it using samples collected in fasting.
  • Question
    Q.How long does the plate coated biotinylated antibody become stable for use?
  • Answer
    A.We have confirmed that it is stable for a week.
  • Question
    Q.How long does streptavidin plate become stable for use after first opening the pack?
  • Answer
    A.We have confirmed that it is stable at 37℃ for 2 weeks.
99999
99999 ELISA - IBL common FAQ
  • Question
    Q.Is composition of EIA buffer of each ELISA kit all same? Can it be mixed to use?
  • Answer
    A.No it isn't. As constitute of each EIA buffer is different, it cannot be mixed with other lots or EIA buffers contained in other kind of ELISA kits.
  • Question
    Q.What is the composition of concentrated wash buffer?
  • Answer
    A.It contains ordinary Tween and phosphate buffer (0.05% Tween-20 in PB).
  • Question
    Q.What is the feature of the plate?
  • Answer
    A.We use plate that is flat bottom and removable strip type plate (8wellx 12 strips).
  • Question
    Q.Can I re-use standard after reconstitution?
  • Answer
    A.Not recommended to re-use standard after reconstitution. Please use it at once after the reconstitution.
  • Question
    Q.What is different between reagent blank and test sample blank?
  • Answer
    A.Reagent blank means a well is only added EIA buffer and the purpose is confirming whether the measuring value is influenced by lack of washing process. Test sample blank means a well is added EIA buffer and HRP antibody and the purpose is to calculate the background.
  • Question
    Q.How many samples can be measured by this kit?
  • Answer
    A.The pre-coated plate contained in our ELISA kit is 96 wells plate. We recommend to use 16 wells (2 slits) for standard and 80 wells (10 slits) for 40 samples in duplicate.
  • Question
    Q.What is the condition of DPP 4 inhibitor?
  • Answer
    A.Only EDTA-plasma collected using collecting tubes contained DPP4 inhibitor can be used for measuring of GLP-1 active form. BD™ P800 GLP-1, GIP, Glucagon, Ghrelin storing vacuumed collecting tube should be used for collecting blood or the blood should be blocked its activity by DPP4 inhibitor prior it is measured. The amount of DPP4 inhibitor for use is 10μL per 1mL blood or 20μL per 1mL plasma.
  • Question
    Q.What is recommended dilution ratio for blood samples?
  • Answer
    A.Please start to dilute from 2 fold.
  • Question
    Q.What is recommended dilution ratio for human samples of healthy subject?
  • Answer
    A.It is 2 to 4 fold.
  • Question
    Q.What size of oligomer can be detected by this assay kit?
  • Answer
    A.Any oligomer that is bigger than Amyloidβ dimer and has a recognized region of 82E1 (N-terminal of 1-16) can be detected by this assay kit.
  • Question
    Q.Can this kit detect protofibril and fibril?
  • Answer
    A.It cross reacts with both protofibril and fibril.
  • Question
    Q.3 recognized parts of 82E1 antibody (a.a. 1-16) are existed if the sample is trimer. In that case, does the conjugated antibody binds to 2parts or only one part?
  • Answer
    A.82E1 antibody is used for as capture antibody (precoated one) and detection antibody (HRP conjugated one). Theoretically, 2 regions of trimer can bind to the detection antibody if 1 region is captured by captured antibody. However, this is only the case that actual molecule detected holds the N-terminal region of Aβ1-16.
  • Question
    Q.Is it depending on the size of oligomer to determine the quantity of the detection antibody that binds to one oligomer?
  • Answer
    A.It depends on molecule of oligomer. It does not detect monomer. We recommend to use both products #27718 Human Amyloidβ (1-40) (FL) Assay Kit - IBL and #27719 Human Amyloidβ (1-42) (FL) Assay Kit - IBL by depending on purposes.
  • Question
    Q.What is recommended dilution ratio for serum samples?
  • Answer
    A.It is 2 to 8 fold.
  • Question
    Q.Can you verify that it is HIV or HBV negative?
  • Answer
    A.The plasma used as the material was verified as negative and not cross reactive against HbsAG, anti-HCV, anti-HBc by the method of approved by FDA.
  • Question
    Q.What is used for the standard?
  • Answer
    A.It is ApoB 100 that is purified from human plasma.