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ELISA - IBL

  • Question
    Q.Do you have any protocol for preparation of mice and/or rat brain
    tissue extracts?
  • Answer
    A.CHAPS is recommended for extractin of wild type (W.T.) mice or rat brain tissue.
    Please refer to the following link for the brain tissue extraction process using CHAPS.
    http://www.ibl-japan.co.jp/files/topics/4944_ext_02_0.pdf

    Guanidine solution is recommended for extraction of Transgenic (Tg) mice or rat brain tissue.
    Please refer to the following link and the following two scientific articles for brain tissue extraction process using Guanidine.
    http://www.ibl-japan.co.jp/files/topics/4945_ext_02_0.pdf

    Presenilin-2 mutation causes early amyloid accumulation and memory impairment in a transgenic mouse model of Alzheimer's disease. Toda T et al. J Biomed Biotechnol. 2011;2011:617974.
    https://www.ncbi.nlm.nih.gov/pubmed/?term=21234330

    Methylene blue modulates β-secretase, reverses cerebral amyloidosis, and improves cognition in transgenic mice. Mori T et al. J Biol Chem. 2014 Oct 31;289(44):30303-17.
    https://www.ncbi.nlm.nih.gov/pubmed/?term=25157105

    * The ELISA kit #27712 Aβ42 and #27714 Aβ40 have been upgraded to #27719 Aβ (1-42)(FL) and #27718 Aβ (1-40)(FL).
  • Question
    Q.Do you have any protocol for preparation of mice and/or rat brain
    tissue extracts?
  • Answer
    A.CHAPS is recommended for extractin of wild type (W.T.) mice or rat brain tissue.
    Please refer to the following link for the brain tissue extraction process using CHAPS.
    http://www.ibl-japan.co.jp/files/topics/4944_ext_02_0.pdf

    Guanidine solution is recommended for extraction of Transgenic (Tg) mice or rat brain tissue.
    Please refer to the following link and the following two scientific articles for brain tissue extraction process using Guanidine.
    http://www.ibl-japan.co.jp/files/topics/4945_ext_02_0.pdf

    Presenilin-2 mutation causes early amyloid accumulation and memory impairment in a transgenic mouse model of Alzheimer's disease. Toda T et al. J Biomed Biotechnol. 2011;2011:617974.
    https://www.ncbi.nlm.nih.gov/pubmed/?term=21234330

    Methylene blue modulates β-secretase, reverses cerebral amyloidosis, and improves cognition in transgenic mice. Mori T et al. J Biol Chem. 2014 Oct 31;289(44):30303-17.
    https://www.ncbi.nlm.nih.gov/pubmed/?term=25157105

    * The ELISA kit #27712 Aβ42 and #27714 Aβ40 have been upgraded to #27719 Aβ (1-42)(FL) and #27718 Aβ (1-40)(FL).
  • Question
    Q.Do you have any protocol for preparation of mice and/or rat brain
    tissue extracts?
  • Answer
    A.CHAPS is recommended for extractin of wild type (W.T.) mice or rat brain tissue.
    Please refer to the following link for the brain tissue extraction process using CHAPS.
    http://www.ibl-japan.co.jp/files/topics/4944_ext_02_0.pdf

    Guanidine solution is recommended for extraction of Transgenic (Tg) mice or rat brain tissue.
    Please refer to the following link and the following two scientific articles for brain tissue extraction process using Guanidine.
    http://www.ibl-japan.co.jp/files/topics/4945_ext_02_0.pdf

    Presenilin-2 mutation causes early amyloid accumulation and memory impairment in a transgenic mouse model of Alzheimer's disease. Toda T et al. J Biomed Biotechnol. 2011;2011:617974.
    https://www.ncbi.nlm.nih.gov/pubmed/?term=21234330

    Methylene blue modulates β-secretase, reverses cerebral amyloidosis, and improves cognition in transgenic mice. Mori T et al. J Biol Chem. 2014 Oct 31;289(44):30303-17.
    https://www.ncbi.nlm.nih.gov/pubmed/?term=25157105

    * The ELISA kit #27712 Aβ42 and #27714 Aβ40 have been upgraded to #27719 Aβ (1-42)(FL) and #27718 Aβ (1-40)(FL).
  • Question
    Q.Do you have any protocol for preparation of mice and/or rat brain
    tissue extracts?
  • Answer
    A.CHAPS is recommended for extractin of wild type (W.T.) mice or rat brain tissue.
    Please refer to the following link for the brain tissue extraction process using CHAPS.
    http://www.ibl-japan.co.jp/files/topics/4944_ext_02_0.pdf

    Guanidine solution is recommended for extraction of Transgenic (Tg) mice or rat brain tissue.
    Please refer to the following link and the following two scientific articles for brain tissue extraction process using Guanidine.
    http://www.ibl-japan.co.jp/files/topics/4945_ext_02_0.pdf

    Presenilin-2 mutation causes early amyloid accumulation and memory impairment in a transgenic mouse model of Alzheimer's disease. Toda T et al. J Biomed Biotechnol. 2011;2011:617974.
    https://www.ncbi.nlm.nih.gov/pubmed/?term=21234330

    Methylene blue modulates β-secretase, reverses cerebral amyloidosis, and improves cognition in transgenic mice. Mori T et al. J Biol Chem. 2014 Oct 31;289(44):30303-17.
    https://www.ncbi.nlm.nih.gov/pubmed/?term=25157105

    * The ELISA kit #27712 Aβ42 and #27714 Aβ40 have been upgraded to #27719 Aβ (1-42)(FL) and #27718 Aβ (1-40)(FL).
  • Question
    Q.What is the epitope of antibodies used in this kit?
  • Answer
    A.Anti-Human Mac-2bp (8A2) Mouse IgG (Precoated Antibody) recognizes position no. 182-276 amino acid of human Mac-2bp.
    Anti- Human Mac-2bp (67A1) Mouse IgG (HRP Antibody) recognizes position no. 361-440 amino acid of human Mac-bp.
  • Question
    Q.What is the immunogen of 2 antibodies used in the kit?
  • Answer
    A.Recombinant protein was used as the immunogen for both antibodies.
  • Question
    Q.Can this kit be used for mouse or rat samples?
  • Answer
    A.No, it cannot.
    A monoclonal antibody (82E1) which is used as the labeled antibody in this kit, specifically reacts to N-terminal of Human Aβ.
    Since the N-terminal sequence of Aβ is different between human and rodents (mouse or rat), it does not cross reacts to mouse and/or rat Aβ.
  • Question
    Q.Can this kit be used for bovine samples?
  • Answer
    A.No, it cannot.
  • Question
    Q.Can this kit be used for bovine samples?
  • Answer
    A.No, it cannot.
  • Question
    Q.Does this kit detect Insulin I or Insulin II?
  • Answer
    A.This kit can be used for quantitatively measuring total insulin including Insulin I, Insulin II and Proinsulin.
  • Question
    Q.Does Neuramidase treatment require for the sample?
  • Answer
    A.No it doesn't. There is no much different for the result between with and without Neuramidase.
  • Question
    Q.I found the recommended dilution ratio is between 200 and 800 fold. That is quite a wide range. What is the most optimized dilution ratio?
  • Answer
    A.Some samples have a very high value of Gd-IgA1 and such samples would be range over that why the recommended dilution ratio is wide.
    We recommend to start from 200 fold dilution and find the best dilution ratio for your samples. The optimized dilution ratio should be determined by each user.
  • 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.Does this kit cross reacts to mouse sAPPα if TG mice that express human sAPPα is used for the assay?
  • Answer
    A.No, it does not cross react to mouse sAPPα. This kit only detects human sAPPα.
  • 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 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 (1-42)?
  • Answer
    A.No it doesn't.
  • Question
    Q.Do you have any protocol for preparation of mice and/or rat brain
    tissue extracts?
  • Answer
    A.CHAPS is recommended for extractin of wild type (W.T.) mice or rat brain tissue.
    Please refer to the following link for the brain tissue extraction process using CHAPS.
    http://www.ibl-japan.co.jp/files/topics/4944_ext_02_0.pdf

    Guanidine solution is recommended for extraction of Transgenic (Tg) mice or rat brain tissue.
    Please refer to the following link and the following two scientific articles for brain tissue extraction process using Guanidine.
    http://www.ibl-japan.co.jp/files/topics/4945_ext_02_0.pdf

    Presenilin-2 mutation causes early amyloid accumulation and memory impairment in a transgenic mouse model of Alzheimer's disease. Toda T et al. J Biomed Biotechnol. 2011;2011:617974.
    https://www.ncbi.nlm.nih.gov/pubmed/?term=21234330

    Methylene blue modulates β-secretase, reverses cerebral amyloidosis, and improves cognition in transgenic mice. Mori T et al. J Biol Chem. 2014 Oct 31;289(44):30303-17.
    https://www.ncbi.nlm.nih.gov/pubmed/?term=25157105

    * The ELISA kit #27712 Aβ42 and #27714 Aβ40 have been upgraded to #27719 Aβ (1-42)(FL) and #27718 Aβ (1-40)(FL).
  • Question
    Q.Does it cross react to mouse or rat Aβ (1-40)?
  • Answer
    A.No it doesn't.
  • Question
    Q.What is recommended dilution ratio for serum samples?
  • Answer
    A.It is 2 to 8 fold.
  • Question
    Q.Can this kit specifically measure phosphorylated α-Synuclein?
  • Answer
    A.No, this kit can measure any type of α-Synuclein. It is not specific for phsphorylated form.
  • 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 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.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.Can rat sample be used for this kit?
  • Answer
    A.No, it cannot.
  • Question
    Q.Is stability data available?
  • Answer
    A.It is available on the following relevant product page as images.
    http://www.ibl-japan.co.jp/en/search/product/detail/id=4678
  • Question
    Q.Can this kit measure mouse blood sample?
  • Answer
    A.No, it cannot.
  • Question
    Q.How can we collect mouse urine sample?
  • Answer
    A.The method of collection of mouse urine is listed on the relevant product page.
    Please refer to the Pamphlet 2 listed on the page.
    http://www.ibl-japan.co.jp/en/search/product/detail/id=4678
  • Question
    Q.How mouse urine sample can be stored?
  • Answer
    A.It should be stored at -80℃ immediately after collection.