Overview

  • Product name

    Mouse Progranulin ELISA Kit
    See all Granulin kits
  • Detection method

    Colorimetric
  • Precision

    Intra-assay
    Sample n Mean SD CV%
    Serum 8 3.3%
    Inter-assay
    Sample n Mean SD CV%
    Serum 3 6.6%
  • Sample type

    Cell culture supernatant, Serum, Cell culture extracts, Hep Plasma, EDTA Plasma, Cit plasma
  • Assay type

    Sandwich (quantitative)
  • Sensitivity

    7.8
  • Range

    93.73 pg/ml - 6000 pg/ml
  • Recovery

    Sample specific recovery
    Sample type Average % Range
    Serum 116 115% - 118%
    Cell culture extracts 107 102% - 109%
    Cell culture media 93 93% - 95%
    Hep Plasma 111 107% - 118%
    EDTA Plasma 115 114% - 118%
    Cit plasma 115 110% - 119%

  • Assay time

    1h 30m
  • Assay duration

    One step assay
  • Species reactivity

    Reacts with: Mouse
    Does not react with: Cow
  • Product overview

    Progranulin in vitro SimpleStep ELISA® (Enzyme-Linked Immunosorbent Assay) kit is designed for the quantitative measurement of mouse Progranulin protein in serum, plasma, cell culture supernatant, and cell extract samples.


    The SimpleStep ELISA® employs an affinity tag labeled capture antibody and a reporter conjugated detector antibody which immunocapture the sample analyte in solution. This entire complex (capture antibody/analyte/detector antibody) is in turn immobilized via immunoaffinity of an anti-tag antibody coating the well. To perform the assay, samples or standards are added to the wells, followed by the antibody mix. After incubation, the wells are washed to remove unbound material. TMB substrate is added and during incubation is catalyzed by HRP, generating blue coloration. This reaction is then stopped by addition of Stop Solution completing any color change from blue to yellow. Signal is generated proportionally to the amount of bound analyte and the intensity is measured at 450 nm. Optionally, instead of the endpoint reading, development of TMB can be recorded kinetically at 600 nm.


    Sensitivity:
    Samples in Sample Diluent NS: 49.5 pg/mL
    Samples in 1X Cell Extraction Buffer PTR: 7.8 pg/mL

  • Notes

    Mouse Progranulin belongs to the granulin family and is 572 amino acids (aa) in length. Additionally, mouse Progranulin is cleaved into individual granulin peptides (Granulin-1 through Granulin-7). Granulins have cytokine-like activity and may also play a role in inflammation, wound repair, and tissue remodelling. They also act as an autocrine growth factor for PC cells. Human and rat Progranulin have 75% and 87% sequence homology with mouse Progranulin, respectively.

  • Tested applications

    Suitable for: Sandwich ELISAmore details
  • Platform

    Pre-coated microplate (12 x 8 well strips)

Properties

  • Storage instructions

    Store at +4°C. Please refer to protocols.
  • Components 1 x 96 tests
    10X Mouse Progranulin Capture Antibody 1 x 600µl
    10X Mouse Progranulin Detector Antibody 1 x 600µl
    10X Wash Buffer PT (ab206977) 1 x 20ml
    50X Cell Extraction Enhancer Solution (ab193971) 1 x 1ml
    5X Cell Extraction Buffer PTR (ab193970) 1 x 10ml
    Antibody Diluent CPR 1 x 6ml
    Mouse Progranulin Lyophilized Recombinant Protein 2 vials
    Plate Seals 1 unit
    Sample Diluent NS (ab193972) 1 x 50ml
    SimpleStep Pre-Coated 96-Well Microplate (ab206978) 1 unit
    Stop Solution 1 x 12ml
    TMB Development Solution 1 x 12ml
  • Research areas

  • Function

    Granulins have possible cytokine-like activity. They may play a role in inflammation, wound repair, and tissue remodeling.
    Granulin-4 promotes proliferation of the epithelial cell line A431 in culture while granulin-3 acts as an antagonist to granulin-4, inhibiting the growth.
  • Tissue specificity

    In myelogenous leukemic cell lines of promonocytic, promyelocytic, and proerythroid lineage, in fibroblasts, and very strongly in epithelial cell lines. Present in inflammatory cells and bone marrow. Highest levels in kidney.
  • Involvement in disease

    Defects in GRN are the cause of ubiquitin-positive frontotemporal dementia (UP-FTD) [MIM:607485]; also known as tau-negative frontotemporal dementia linked to chromosome 17. Frontotemporal dementia (FTD) is the second most common cause of dementia in people under the age of 65 years. It is an autosomal dominant neurodegenerative disease.
  • Sequence similarities

    Belongs to the granulin family.
  • Post-translational
    modifications

    Granulins are disulfide bridged.
  • Cellular localization

    Secreted.
  • Information by UniProt
  • Alternative names

    • Acrogranin
    • CLN11
    • GEP
    • GP88
    • Granulin A
    • Granulin B
    • Granulin C
    • Granulin D
    • Granulin E
    • Granulin epithelin
    • Granulin F
    • Granulin G
    • Granulin-7
    • Granulins
    • GRN
    • GRN_HUMAN
    • PC cell derived growth factor
    • PCDGF
    • PEPI
    • PGRN
    • Proepithelin
    • Progranulin
    see all
  • Database links

Applications

Our Abpromise guarantee covers the use of ab213473 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Application Abreviews Notes
Sandwich ELISA Use at an assay dependent concentration.

Images

  • SimpleStep ELISA technology allows the formation of the antibody-antigen complex in one single step, reducing assay time to 90 minutes. Add samples or standards and antibody mix to wells all at once, incubate, wash, and add your final substrate. See protocol for a detailed step-by-step guide.

     

  • Background-subtracted data values (mean +/- SD) are graphed.

  • Background-subtracted data values (mean +/- SD) are graphed.

  • The concentrations of Progranulin were measured in duplicates, interpolated from the Progranulin standard curves and corrected for sample dilution. Undiluted samples are as follows: serum 1:250, plasma (citrate) 1:250, plasma (EDTA) 1:250, and plasma (heparin) 1:250. The interpolated dilution factor corrected values are plotted (mean +/- SD, n=2). The mean Progranulin concentration was determined to be 1,610.5 ng/mL in serum, 1,149.5 ng/mL in plasma (citrate), 1,253.7 ng/mL in plasma (EDTA) and 1,710.8 ng/mL in plasma (heparin).

  • The concentrations of Progranulin were measured in duplicates, interpolated from the Progranulin standard curves and corrected for sample dilution. Undiluted samples are as follows: J774A.1 cultured media 1:400, RAW264.7 cultured media 6.25%, and RPMI culture media 10%. The interpolated dilution factor corrected values are plotted (mean +/- SD, n=2). The mean Progranulin concentration was determined to be 197.4 ng/mL in J774A.1 cultured media, 38.4 ng/mL in RAW264.7 cultured media, and 82.6 ng/mL in RPMI culture media. J774A.1 samples were cultured in high glucose DMEM culture media with kanamycin and 10% fetal bovine serum for 72 hours and then stimulated with 1.5% PHA plus 10 ng/mL PMA for 72 hours. RAW264.7 samples were cultured in high glucose DMEM culture media with kanamycin and L‑Glutamine for 72 hours and then stimulated with 5 µg/mL of LPS for 48 hours.

  • The concentrations of Progranulin were measured in duplicate and interpolated from the Progranulin standard curve and corrected for sample dilution. The interpolated dilution factor corrected values are plotted (mean +/- SD, n=2). The mean Progranulin concentration in J774A.1 cell extract was determined to be 2.9 ng/mL.

  • All samples were diluted 1:400 and the concentrations of Progranulin were measured in duplicates, interpolated from the Progranulin standard curves and corrected for sample dilution. The interpolated dilution factor corrected values are plotted (mean +/- SD, n=2). The mean Progranulin concentration was determined to be 34.8 ng/mL in unstimulated J774A.1 cultured media and 206.9 ng/mL in stimulated J774A.1 cultured media. J774A.1 samples were cultured in high glucose DMEM culture media with kanamycin and 10% fetal bovine serum for 72 hours (unstimulated samples) and then stimulated with 1.5% PHA plus 10 ng/mL PMA for 72 hours.

Protocols

References

ab213473 has not yet been referenced specifically in any publications.

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