Mature NGF/proNGF Combo (BEK-2212/2226), Human, Rapid™ ELISA assay

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Catalog Number
BEK-2223

    Product Info

  • Product Name Mature NGF/proNGF Combo (BEK-2212/2226), Human, Rapid™ ELISA assay
  • Product Description google The Biosensis Human Mature NGF/proNGF Combo RapidTM enzyme-linked immunosorbent assay (ELISA) Kit combines individual, but complementary ELISA kits for the two most important NGF isoforms: Mature NGF (BEK-2212) and full-length proNGF (BEK-2226).

    Both kits are sandwich ELISAs, useful for the quantification of mature NGF and proNGF in less than 4 hours in cell culture supernatants, serum, and plasma (citrate) only if used as directed, with a simplified protocol and no loss of sensitivity or specificity. Researchers have used both kits for NGF/proNGF measurement in human urine, however, Biosensis has not yet internally validated the use of urine in both ELISA kits. End-users are strongly advised to perform essential validation experiments to assure accurate NGF/proNGF quantification in human urine. Please refer to the most current kit protocols for BEK-2212 (Mature BDNF RapidTM ELISA) and BEK-2226 (proNGF RapidTM ELISA), for specific use instructions for each substrate application, in particular blood samples.

    The Mature NGF ELISA kit consists of a pre-coated mouse monoclonal anti-NGF capture antibody, a biotinylated anti-NGF detection antibody and horseradish peroxidase (HRP)-conjugated streptavidin. The proNGF ELISA kit consists of a pre-coated anti-proNGF capture antibody, a biotinylated anti-proNGF detection antibody and horseradish peroxidase (HRP)-conjugated streptavidin. The addition of a substrate (3,3',5,5'-tetramethylbenzidine, TMB) yields a coloured reaction product which is directly proportional to the concentration of mature NGF or proNGF present in samples and protein standards. A NGF and proNGF positive control (QC sample) is provided to assure consistent assay performance.

    The Mature NGF ELISA kit employs a high-quality recombinant human NGF standard approved by the World Health Organization (WHO). The proNGF ELISA kit contains a recombinant human proNGF standard expressed in E.coli. Note that this Mature NGF/proNGF Combo kit is designed to measure the human protein forms, although due to sequence homology the Mature NGF ELISA kit will detect mouse and rat mature NGF. The proNGF ELISA kit does not cross-react with rodent forms of proNGF.

    This Combo kit is capable of distinguishing and independently quantifying the mature NGF and full-length NGF isoforms. Internal validation data demonstrates <0.1% cross-reactivity (determined at 25 ng/mL, diluted in assay buffer) of human proNGF protein in the mature NGF ELISA, demonstrating the preferential quantification of mature NGF over full-length human proNGF. The absence of proNGF cross-reactivity in the mature NGF ELISA kit has been independently confirmed by Mossa AH et al. (2020). The antibodies used in the proNGF ELISA kit bind epitopes within the pro-domain of the protein and therefore recognize proNGF and the pro-domain peptide, but do not cross-react with mature NGF!

    Important: Accurate NGF quantification in human citrate-plasma requires the addition of Heterophilic Antibody Blocker BL-005-500. Accurate proNGF quantification in human serum requires the addition of Heterophilic Antibody Blocker BL-003-1000. Both Heterophilic Antibody Blockers are provided in the kit.

    This ELISA kit has not been tested for other applications. It has been configured for research use only and is not to be used for diagnostic or clinical procedures.
  • Alternative Names Beta-nerve growth factor; Ngfb; NGF; pro-beta nerve growth factor; proNGF
  • Application(s) ELISA
  • Specificity Mature NGF ELISA:
    Detects human NGF, and cross-reacts with mouse and rat mature NGF. This mature NGF ELISA preferentially detects mature NGF over full-length proNGF.

    proNGF ELISA:
    Detects human proNGF only, does not cross-react with mouse and rat proNGF. Both capture and detection antibodies used in the proNGF ELISA kit binds to epitopes within the pro-domain of proNGF. Thus, the proNGF ELISA detects the full-length form of proNGF, and does not quantify mature NGF.
  • Species Reactivity Human
  • Immunogen Description See BEK-2212 & BEK-2226 for specific details
  • Range Mature NGF: 3.9 – 250 pg/mL
    proNGF: 0.078 – 5 ng/mL
  • Sensitivity Typical limit of detection (LOD) for human mature NGF is 2 pg/mL determined as 150% of the blank value. Typical limit of detection (LOD) for human proNGF is < 60 pg/mL, determined as blank value plus 3x standard deviation of blank OD (n=10).
  • Regulatory Status For research use only.

    Specifications

  • Product Description The Biosensis Human Mature NGF/proNGF Combo RapidTM enzyme-linked immunosorbent assay (ELISA) Kit combines individual, but complementary ELISA kits for the two most important NGF isoforms: Mature NGF (BEK-2212) and full-length proNGF (BEK-2226).

    Both kits are sandwich ELISAs, useful for the quantification of mature NGF and proNGF in less than 4 hours in cell culture supernatants, serum, and plasma (citrate) only if used as directed, with a simplified protocol and no loss of sensitivity or specificity. Researchers have used both kits for NGF/proNGF measurement in human urine, however, Biosensis has not yet internally validated the use of urine in both ELISA kits. End-users are strongly advised to perform essential validation experiments to assure accurate NGF/proNGF quantification in human urine. Please refer to the most current kit protocols for BEK-2212 (Mature BDNF RapidTM ELISA) and BEK-2226 (proNGF RapidTM ELISA), for specific use instructions for each substrate application, in particular blood samples.

    The Mature NGF ELISA kit consists of a pre-coated mouse monoclonal anti-NGF capture antibody, a biotinylated anti-NGF detection antibody and horseradish peroxidase (HRP)-conjugated streptavidin. The proNGF ELISA kit consists of a pre-coated anti-proNGF capture antibody, a biotinylated anti-proNGF detection antibody and horseradish peroxidase (HRP)-conjugated streptavidin. The addition of a substrate (3,3',5,5'-tetramethylbenzidine, TMB) yields a coloured reaction product which is directly proportional to the concentration of mature NGF or proNGF present in samples and protein standards. A NGF and proNGF positive control (QC sample) is provided to assure consistent assay performance.

    The Mature NGF ELISA kit employs a high-quality recombinant human NGF standard approved by the World Health Organization (WHO). The proNGF ELISA kit contains a recombinant human proNGF standard expressed in E.coli. Note that this Mature NGF/proNGF Combo kit is designed to measure the human protein forms, although due to sequence homology the Mature NGF ELISA kit will detect mouse and rat mature NGF. The proNGF ELISA kit does not cross-react with rodent forms of proNGF.

    This Combo kit is capable of distinguishing and independently quantifying the mature NGF and full-length NGF isoforms. Internal validation data demonstrates <0.1% cross-reactivity (determined at 25 ng/mL, diluted in assay buffer) of human proNGF protein in the mature NGF ELISA, demonstrating the preferential quantification of mature NGF over full-length human proNGF. The absence of proNGF cross-reactivity in the mature NGF ELISA kit has been independently confirmed by Mossa AH et al. (2020). The antibodies used in the proNGF ELISA kit bind epitopes within the pro-domain of the protein and therefore recognize proNGF and the pro-domain peptide, but do not cross-react with mature NGF!

    Important: Accurate NGF quantification in human citrate-plasma requires the addition of Heterophilic Antibody Blocker BL-005-500. Accurate proNGF quantification in human serum requires the addition of Heterophilic Antibody Blocker BL-003-1000. Both Heterophilic Antibody Blockers are provided in the kit.

    This ELISA kit has not been tested for other applications. It has been configured for research use only and is not to be used for diagnostic or clinical procedures.
  • Related Products Nerve growth factor (Beta-NGF), Human, Rapid™ ELISA assay
    Nerve growth factor, pro- (proNGF), Human, Rapid™ ELISA assay
  • Application(s) ELISA
  • Application Details ELISA. For the quantification of Mature NGF and proNGF in Culture Supernatant, Serum, Plasma (Citrate). Please download the detailed product insert for complete instructions for the successful use of this ELISA. Use only as directed.
  • Target Mature NGF/proNGF Combo (BEK-2212/2226)
  • Specificity Mature NGF ELISA:
    Detects human NGF, and cross-reacts with mouse and rat mature NGF. This mature NGF ELISA preferentially detects mature NGF over full-length proNGF.

    proNGF ELISA:
    Detects human proNGF only, does not cross-react with mouse and rat proNGF. Both capture and detection antibodies used in the proNGF ELISA kit binds to epitopes within the pro-domain of proNGF. Thus, the proNGF ELISA detects the full-length form of proNGF, and does not quantify mature NGF.
  • Target Cross-Reactivity (ELISA) Cross-reactivity of NGF isoforms:

    Mature NGF ELISA:
    The antibodies used in the Mature NGF ELISA kit bind epitopes within the mature domain of the protein. However, human proNGF protein shows <0.1% cross-reactivity (determined at 25 ng/mL, diluted in assay buffer), demonstrating the preferential quantification of mature NGF over full-length human proNGF. The absence of proNGF cross-reactivity in the mature NGF ELISA kit has been independently confirmed by Mossa AH et al. (2020). The Mature NGF ELISA does not cross-react with brain derived neurotrophic factor (BDNF), neurotrophin-3 (NT-3), NT-4/5, glial cell line-derived neurotrophic factor (GDNF) and vascular endothelial growth factor (VEGF165) tested at 25 ng/mL. Due to a high degree of NGF sequence homology, the antibodies used in this kit will also detect mature NGF from numerous other species including mouse and rat!

    proNGF ELISA:
    Does not cross-react with recombinant human mature NGF and proBDNF tested at 25 ng/mL. Does not react with rodent proNGF.
  • Target Host Species Human
  • Species Reactivity Human
  • Immunogen Description See BEK-2212 & BEK-2226 for specific details
  • Range Mature NGF: 3.9 – 250 pg/mL
    proNGF: 0.078 – 5 ng/mL
  • Sensitivity Typical limit of detection (LOD) for human mature NGF is 2 pg/mL determined as 150% of the blank value. Typical limit of detection (LOD) for human proNGF is < 60 pg/mL, determined as blank value plus 3x standard deviation of blank OD (n=10).
  • Sample Type Culture Supernatant, Plasma (Citrate), Serum
  • Detection Method Colorimetric
  • Kit Components The ELISA kit box contains 2 x 96-well pre-coated strip plates per kit (1 x NGF antibody, 1 x proNGF antibody coated plate), protein standards, QC sample, detection reagents, heterophilic antibody blocker, wash and sample buffers, substrate buffer and detailed protocols.
  • Storage Instructions Store at 2-8°C
  • Storage Temperature 2-8°C
  • Batch Number Please see item label.
  • Expiration Date 12 months after date of receipt if unopened.
  • Alternative Names Beta-nerve growth factor; Ngfb; NGF; pro-beta nerve growth factor; proNGF
  • Uniprot Number P01138
  • Uniprot Number/Name P01138 (NGF_HUMAN)
  • Scientific Background Nerve growth factor (NGF) is synthesized as a precursor (proNGF) which may be released and have physiological functions to cause cell death. It binds neurotrophin receptor p75 and sortilin and may also be important for the development of nervous system. proNGF is synthesized in target tissues and glia, transported retrogradely and may be released.
  • Research Area Neurotrophins; Cancer
  • Shipping Statement For order quantities larger than 1 plate, this product is supplied in package units of 2 plates.
  • Shipping Temperature 2-8°C (on cold packs)
  • UNSPSC CODE 41116158
  • Regulatory Status For research use only.