Overview

  • Product name

    Human FLRG ELISA Kit
    See all FSTL3 kits
  • Detection method

    Colorimetric
  • Precision

    Intra-assay
    Sample n Mean SD CV%
    serum 8 4.7%
    Inter-assay
    Sample n Mean SD CV%
    serum 3 6.4%
  • Sample type

    Cell culture supernatant, Milk, Urine, Serum, Hep Plasma, EDTA Plasma
  • Assay type

    Sandwich (quantitative)
  • Sensitivity

    8.2 pg/ml
  • Range

    31.25 pg/ml - 2000 pg/ml
  • Recovery

    Sample specific recovery
    Sample type Average % Range
    Cell culture supernatant 107 104% - 111%
    Milk 103 98% - 109%
    Urine 98 94% - 109%
    Serum 106 105% - 109%
    Hep Plasma 101 99% - 105%
    EDTA Plasma 104 102% - 106%

  • Assay time

    1h 30m
  • Assay duration

    One step assay
  • Species reactivity

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

    FLRG in vitro SimpleStep ELISA® (Enzyme-Linked Immunosorbent Assay) kit is designed for the quantitative measurement of FLRG protein in human serum, plasmas, urine, milk, and cell culture supernatants.


    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 Development Solution 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.


    FLRG (follistatin-related gene protein), also known as Follistatin-like 3 (FSTL3) and follistatin-related protein (FSRP) is an activin-binding glycoprotein encoded by the gene FSTL3. FLRG is a 237- amino acid glycoprotein, that contains a TGF-beta binding domain, two Follistatin-like, and two Kazal-like domains. FLRG inhibits activin-A, activin-B, BMP2-, and MSDT- induced cellular signaling. FLRG is highly expressed in placental tissue, testis, and the adrenal gland. An isoform of FLRG that lacks the signal sequence is located in the nucleus and has been found to bind to the transcription factor AF10, which enhances AF10-dependant gene transcription. FLRG is involved in a wide range of functions including bone formation, inhibition of osteoclast differentiation, hematopoiesis, and increases cell hematopoietic cell adhesion to fibronectin.    

  • 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 Human FLRG Capture Antibody 1 x 600µl
    10X Human FLRG Detector Antibody 1 x 600µl
    10X Wash Buffer PT (ab206977) 1 x 20ml
    Antibody Diluent 5BI 1 x 6ml
    Human FLRG 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

    Isoform 1 or the secreted form is a binding and antagonizing protein for members of the TGF-beta family, such us activin, BMP2 and MSTN. Inhibits activin A-, activin B-, BMP2- and MSDT-induced cellular signaling; more effective on activin A than on activin B. Involved in bone formation; inhibits osteoclast differentiationc. Involved in hematopoiesis; involved in differentiation of hemopoietic progenitor cells, increases hematopoietic cell adhesion to fibronectin and seems to contribute to the adhesion of hematopoietic precursor cells to the bone marrow stroma. Isoform 2 or the nuclear form is probably involved in transcriptional regulation via interaction with MLLT10.
  • Tissue specificity

    Expressed in a wide range of tissues.
  • Involvement in disease

    Note=A chromosomal aberration involving FSTL3 is found in a case of B-cell chronic lymphocytic leukemia. Translocation t(11;19)(q13;p13) with CCDN1.
  • Sequence similarities

    Contains 2 follistatin-like domains.
    Contains 2 Kazal-like domains.
    Contains 1 TB (TGF-beta binding) domain.
  • Cellular localization

    Secreted and Nucleus. Although alternative initiation has been demonstrated and resulted in different localization, the major source of nuclear FSTL3 appears not to depend on translation initiation at Met-27 according to.
  • Information by UniProt
  • Alternative names

    • FLRG
    • follistatin-like 3
    • follistatin-like 3 glycoprotein
    • Follistatin-like protein 3
    • Follistatin-related gene protein
    • Follistatin-related protein 3
    • FSRP
    • Fstl3
    • FSTL3_HUMAN
    see all
  • Database links

Associated products

Applications

Our Abpromise guarantee covers the use of ab245715 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.

     

  • Standard Curve comparison between human FLRG SimpleStep ELISA kit and traditional ELISA kit from leading competitor. SimpleStep ELISA kit shows increased sensitivity.
  • The FLRG standard curve was prepared as described in Section 10. Raw data values are shown in the table. Background-subtracted data values (mean +/- SD) are graphed.
  • The concentrations of FLRG were measured in duplicates, interpolated from the FLRG standard curves and corrected for sample dilution. Undiluted samples are as follows: serum 15%, plasma (EDTA) 15%, and plasma (heparin) 15%. The interpolated dilution factor corrected values are plotted (mean +/- SD, n=2). The mean FLRG concentration was determined to be 8,804 pg/mL in serum, 8,996 pg/mL in plasma (EDTA) and 9,092 pg/mL in plasma (heparin).
  • The concentrations of FLRG were measured in duplicates, interpolated from the FLRG standard curves and corrected for sample dilution. Undiluted samples are as follows: milk 15% and urine 15%. The interpolated dilution factor corrected values are plotted (mean +/- SD, n=2). The mean FLRG concentration was determined to be 3,167 pg/mL in milk and 6,392 pg/mL in urine.
  • The concentrations of FLRG were measured in duplicates, interpolated from the FLRG standard curves and corrected for sample dilution. Undiluted samples are as follows: JAR cell supernatants 1%. The interpolated dilution factor corrected values are plotted (mean +/- SD, n=2). The mean FLRG concentration was determined to be 83,631 pg/mL in JAR cell supernatant.
  • Interpolated dilution factor corrected values are plotted (mean +/- SD, n=2). The mean FLRG concentration was determined to be 15,897 pg/mL with a range of 13,362 - 21,062 pg/mL.

Protocols

References

ab245715 has not yet been referenced specifically in any publications.

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