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    products/elisa/human-galectin-3-elisa-kit-lgals3-ab269555.pdf

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Validated using a knockout cell lineRecombinantSimpleStep

Human Galectin-3 ELISA Kit (LGALS3) (ab269555)

  • Datasheet
  • SDS
  • Protocol Booklet
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SimpleStep ELISA Protocol Diagram
  • Human Gelectin-3 standard curve comparison
  • Human Gelectin-3 standard curve comparison
  • Example of human Galectin-3 standard curve in Sample Diluent NS.
  • Interpolated concentrations of native Galectin-3 in human serum and plasma samples.
  • Interpolated concentrations of native Galectin-3 in human PBMC cell culture supernatant samples cultured for 36 hours.
  • Interpolated concentrations of native Galectin-3 in human MCF7 cell based on a 1.7 µg/mL extract load, Hap1 wild type and Galectin-3 knockout cells based on 200 µg/mL extract load.
  • Serum from ten individual healthy human female donors was measured in duplicate.
  • Sandwich ELISA - Human Galectin-3 ELISA Kit (LGALS3) (ab269555)

Key features and details

  • One-wash 90 minute protocol
  • Sensitivity: 13.3 pg/ml
  • Range: 58.8 pg/ml - 2000 pg/ml
  • Sample type: Cell culture media, Cell culture supernatant, Cell Lysate, Cit plasma, EDTA Plasma, Hep Plasma, Serum
  • Detection method: Colorimetric
  • Assay type: Sandwich (quantitative)
  • Reacts with: Human

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Overview

  • Product name

    Human Galectin-3 ELISA Kit (LGALS3)
    See all Galectin 3 kits
  • Detection method

    Colorimetric
  • Precision

    Intra-assay
    Sample n Mean SD CV%
    Serum 8 3.4%
    Inter-assay
    Sample n Mean SD CV%
    Serum 3 1.6%
  • Sample type

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

    Sandwich (quantitative)
  • Sensitivity

    13.3 pg/ml
  • Range

    58.8 pg/ml - 2000 pg/ml
  • Recovery

    Sample specific recovery
    Sample type Average % Range
    Cell culture supernatant 114 109% - 120%
    Serum 113 107% - 120%
    Cell Lysate 109 103% - 118%
    Cell culture media 93 91% - 95%
    Hep Plasma 109 107% - 111%
    EDTA Plasma 113 111% - 116%
    Cit plasma 110 106% - 114%
  • Assay time

    1h 30m
  • Assay duration

    One step assay
  • Species reactivity

    Reacts with: Human
  • Product overview

    Human Galectin-3 ELISA Kit (LGALS3) (ab269555) is a single-wash 90 min sandwich ELISA designed for the quantitative measurement of Galectin-3 (LGALS3) protein in cell lysate, serum, cell culture media, cit plasma, edta plasma, hep plasma, and cell culture supernatant. It uses our proprietary SimpleStep ELISA® technology. Quantitate Human Galectin-3 (LGALS3) with 13.3 pg/ml sensitivity.


    SimpleStep ELISA® technology employs capture antibodies conjugated to an affinity tag that is recognized by the monoclonal antibody used to coat our SimpleStep ELISA® plates. This approach to sandwich ELISA allows the formation of the antibody-analyte sandwich complex in a single step, significantly reducing assay time. See the SimpleStep ELISA® protocol summary in the image section for further details. Our SimpleStep ELISA® technology provides several benefits:


            - Single-wash protocol reduces assay time to 90 minutes or less
            - High sensitivity, specificity and reproducibility from superior antibodies
            - Fully validated in biological samples
            - 96-wells plate breakable into 12 x 8 wells strips


    A 384-well SimpleStep ELISA® microplate (ab203359) is available to use as an alternative to the 96-well microplate provided with SimpleStep ELISA® kits.

  • Notes

    Galectin-3 (Gal-3, Galactoside-binding protein, Mac-2 antigen) is a member of the galectin family of carbohydrate-binding proteins. Galectin-3 is active intracellularly and extracellularly. It is widely expressed, including by fibroblasts and macrophages. Intracellularly, Galectin-3 can function in the nucleus as a pre-mRNA splicing factor and in the cytoplasm it has roles in endocytosis, exocytosis and apoptosis. Extracellularly, Galectin-3 is often pro-inflammatory and is involved in innate immunity. Galectin-3 can be monomeric or form higher order homo and hetero oligomers.

    Abcam has not and does not intend to apply for the REACH Authorisation of customers’ uses of products that contain European Authorisation list (Annex XIV) substances.
    It is the responsibility of our customers to check the necessity of application of REACH Authorisation, and any other relevant authorisations, for their intended uses.

  • 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 Galectin-3 (LGALS3) Capture Antibody 1 x 600µl
    10X Human Galectin-3 (LGALS3) Detector Antibody 1 x 600µl
    10X Wash Buffer PT (ab206977) 1 x 20ml
    5X Cell Extraction Buffer PTR (ab193970) 1 x 10ml
    Antibody Diluent 5BI 1 x 6ml
    Human Galectin-3 (LGALS3) Lyophilized Recombinant Protein 2 vials
    Plate Seals 1 unit
    Sample Diluent NS (ab193972) 2 x 50ml
    SimpleStep Pre-Coated 96-Well Microplate (ab206978) 1 unit
    Stop Solution 1 x 12ml
    TMB Development Solution 1 x 12ml
  • Research areas

    • Neuroscience
    • Cell Adhesion Proteins
    • Nuclear Proteins
    • Cell Biology
    • Cell Cycle
    • Cell differentiation
  • Function

    Galactose-specific lectin which binds IgE. May mediate with the alpha-3, beta-1 integrin the stimulation by CSPG4 of endothelial cells migration. Together with DMBT1, required for terminal differentiation of columnar epithelial cells during early embryogenesis.
  • Tissue specificity

    A major expression is found in the colonic epithelium. It is also abundant in the activated macrophages.
  • Sequence similarities

    Contains 1 galectin domain.
  • Cellular localization

    Nucleus. Cytoplasmic in adenomas and carcinomas. May be secreted by a non-classical secretory pathway and associate with the cell surface.
  • Target information above from: UniProt accession P17931 The UniProt Consortium
    The Universal Protein Resource (UniProt) in 2010
    Nucleic Acids Res. 38:D142-D148 (2010) .

    Information by UniProt
  • Alternative names

    • 35 kDa lectin
    • Carbohydrate binding protein 35
    • Carbohydrate-binding protein 35
    • CBP 35
    • CBP35
    • Gal-3
    • GAL3
    • Galactose-specific lectin 3
    • Galactoside-binding protein
    • GALBP
    • Galectin 3 internal gene,included
    • Galectin-3
    • Galectin3
    • GALIG
    • GBP
    • IgE binding protein
    • IgE-binding protein
    • L 31
    • L 34
    • L-31
    • L-34 galactoside-binding lectin
    • L31
    • Laminin-binding protein
    • Lectin L-29
    • Lectin, galactose binding, soluble 3
    • LEG3_HUMAN
    • LGALS2
    • LGALS3
    • MAC 2 antigen
    • Mac-2
    • Mac-2 antigen
    • MAC2
    • Macrophage galactose-specific lectin
    • MGC105387
    see all
  • Database links

    • Entrez Gene: 3958 Human
    • Omim: 153619 Human
    • SwissProt: P17931 Human
    • Unigene: 531081 Human

    Associated products

    • Alternative Versions

      • Human Galectin-3 Antibody Pair - BSA and Azide free (ab270391)

    Images

    • SimpleStep ELISA Protocol Diagram
      SimpleStep ELISA Protocol Diagram

      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.

    • Human Gelectin-3 standard curve comparison
      Human Gelectin-3 standard curve comparison

      Standard Curve comparison between human Galectin-3 SimpleStep ELISA kit and traditional ELISA kit from leading competitor. SimpleStep ELISA kit shows increased sensitivity.

    • Human Gelectin-3 standard curve comparison
      Human Gelectin-3 standard curve comparison

      Standard Curve comparison between the original Human Gelectin-3 SimpleStep ELISA kit (ab188394) and current Human Gelectin-3 SimpleStep ELISA kit (ab269555).

    • Example of human Galectin-3 standard curve in Sample Diluent NS.
      Example of human Galectin-3 standard curve in Sample Diluent NS.

      The Galectin-3 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.

    • Interpolated concentrations of native Galectin-3 in human serum and plasma samples.
      Interpolated concentrations of native Galectin-3 in human serum and plasma samples.

      The concentrations of Galectin-3 were measured in duplicates, interpolated from the Galectin-3 standard curves and corrected for sample dilution. Undiluted samples are as follows: serum 6%, plasma (citrate) 6%, plasma (EDTA) 6% and plasma (heparin) 10%. The interpolated dilution factor corrected values are plotted (mean +/- SD, n=2). The mean Galectin-3 concentration was determined to be 34.5 ng/mL in neat serum, 30.3 ng/mL in neat plasma (citrate), 29.9 ng/mL in neat plasma (EDTA) and 14.8 ng/mL in neat plasma (heparin).

    • Interpolated concentrations of native Galectin-3 in human PBMC cell culture supernatant samples cultured for 36 hours.
      Interpolated concentrations of native Galectin-3 in human PBMC cell culture supernatant samples cultured for 36 hours.

      The concentrations of Galectin-3 were measured in duplicates, interpolated from the Galectin-3 standard curves and corrected for sample dilution. Undiluted samples are as follows: PBMC supernatant 5%. The interpolated dilution factor corrected values are plotted (mean +/- SD, n=2). The mean Galectin-3 concentration was determined to be 17.2 ng/mL in neat PBMC supernatant.

    • Interpolated concentrations of native Galectin-3 in human MCF7 cell based on a 1.7 µg/mL extract load, Hap1 wild type and Galectin-3 knockout cells based on 200 µg/mL extract load.
      Interpolated concentrations of native Galectin-3 in human MCF7 cell based on a 1.7 µg/mL extract load, Hap1 wild type and Galectin-3 knockout cells based on 200 µg/mL extract load.

      The concentrations of Galectin-3 were measured in duplicate and interpolated from the Galectin-3 standard curve and corrected for sample dilution. The interpolated dilution factor corrected values are plotted (mean +/- SD, n=2). The mean Galectin-3 concentration was determined to be 1,855 pg/mL in MCF7 extract, 1,548 pg/mL in Hap1 wild type extract and undetectable in Hap1 Galectin-3 knockout extract.

    • Serum from ten individual healthy human female donors was measured in duplicate.
      Serum from ten individual healthy human female donors was measured in duplicate.

      Interpolated dilution factor corrected values are plotted (mean +/- SD, n=2). The mean Galectin-3 concentration was determined to be 40.4 ng/mL with a range of 27.2 – 66.0 ng/mL.

    • Sandwich ELISA - Human Galectin-3 ELISA Kit (LGALS3) (ab269555)
      Sandwich ELISA - Human Galectin-3 ELISA Kit (LGALS3) (ab269555)
      To learn more about the advantages of recombinant antibodies see here.

    Protocols

    • Protocol Booklet

    Click here to view the general protocols

    Datasheets and documents

    • SDS download

    • Datasheet download

      Download

    References (0)

    Publishing research using ab269555? Please let us know so that we can cite the reference in this datasheet.

    ab269555 has not yet been referenced specifically in any publications.

    Customer reviews and Q&As

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    1-2 of 2 Abreviews

    Unpublished data as of yet

    Excellent Excellent 5/5 (Ease of Use)
    Abreviews
    Abreviews
    This kit worked very well. It was easy to use and follow the directions. We used it for both cell lysates and for cell supernatants. The standard curve worked very well, giving a 0.999 R^2 value. We decided to run one sample at a few different dilutions against the standard the first time in order to optimize the dilution factor necessary to stay within the standard curve. This was well worth using a few well so as to not use the whole plate with samples that were improperly diluted.
    Great results. Many such cases. Recommend for sure.
    The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

    Abcam user community

    Verified customer

    Submitted May 10 2021

    Reactivity of Human Galectin-3 ELISA Kit in Pig plasma samples

    Good Excellent 5/5 (Ease of Use)
    Abreviews
    Abreviews
    abreview image
    We have used your Human Galectin-3 ELISA Kit to quantify Galectin3 levels in pig plasma samples.
    Since very few research groups work with porcine animal models, there are not many kits with specificity to pigs.Therefore, we usually try with human kits due to his huge similarity to human.
    This is why we have tried to test our plasma samples from a atrial fibrillation porcine animal model. According to literature, Galectin3 is upregulated during atrial fibrillation and is released to the blood such a way it can be measured and detected in plasma samples. In our case, it was very interesting to quantify the levels of Galectin3 in plasma samples taken from the inner atrial cavity, since in case that Galectin3 is being released by the atrial cardiomyocytes, the atrial blood pool would have increased Galectin3 levels compared to peripheral blood.
    However, in our experiment with your Human Galectin-3 ELISA Kit we have not been able to detect Gal3 levels in pig plasma samples. We have followed point by point the protocol of your kit, with the recommended dilutions of the Heparin plasma samples, making duplicates for statistical reasons and with several animal conditions to verify differences among different atrial fibrillation animal models.
    In the attached image you can verify as the first 2 columns of the plate (A and B) correspond to the Standard Curve (first 2 wells -A1 and B1- correspond the blank wells and the rest of them are the decreasing concentrations points of the standard curve). The standard curve fits a 4PL curve with an r2=0.997. The rest of the wells are the pig plasma samples and all of them show a coloration deposit similar to the control blank control.
    Measuring the OD450nm the results show that there is no any detection of Gal3 in our plasma samples from pigs.
    Therefore, I would recommend this kit for using it with human samples. It is very easy, you can achieve your results in just one hour and the protocol is very easy to follow. However, it is clear that it is not a proper kit for pig samples since it is not able to detect any Galectin3 in this kind of samples.
    The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

    Abcam user community

    Verified customer

    Submitted Dec 14 2020

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