Human Galectin-3 DuoSet ELISA

Catalog # Availability Size / Price Qty
DY1154
Ancillary Products Available
Human Galectin-3 ELISA Standard Curve
1 Image
Product Details
Procedure
Citations (7)
FAQs
Supplemental Products
Reviews (2)

Human Galectin-3 DuoSet ELISA Summary

Assay Type
Solid Phase Sandwich ELISA
Format
96-well strip plate
Sample Volume Required
100 µL
Sufficient Materials
For fifteen 96-well plates*
Specificity
Please see the product datasheet

* Provided that the recommended microplates, buffers, diluents, substrates and solutions are used, and the assay is run as summarized in the Assay Procedure provided.

This DuoSet ELISA Development kit contains the basic components required for the development of sandwich ELISAs to measure natural and recombinant human Galectin-3. The suggested diluent is suitable for the analysis of most cell culture supernate samples. Diluents for complex matrices, such as serum and plasma, should be evaluated prior to use in this DuoSet ELISA.

Product Features

  • Optimized capture and detection antibody pairings with recommended concentrations save lengthy development time
  • Development protocols are provided to guide further assay optimization
  • Assay can be customized to your specific needs
  • Economical alternative to complete kits

Kit Content

  • Capture Antibody
  • Detection Antibody
  • Recombinant Standard
  • Streptavidin conjugated to horseradish-peroxidase (Streptavidin-HRP)

Other Reagents Required


PBS: (Catalog # DY006), or 137 mM NaCl, 2.7 mM KCl, 8.1 mM Na2HPO4, 1.5 mM KH2PO4, pH 7.2 - 7.4, 0.2 µm filtered

Wash Buffer: (Catalog # WA126), or equivalent

Reagent Diluent: 1% BSA in PBS, pH 7.2-7.4, 0.2 m filtered (R&D Systems Catalog # DY995). Quality of BSA is critical (see Technical Hints).

Blocking Buffer: 1% BSA in PBS, pH 7.2-7.4, 0.2 m filtered (R&D Systems Catalog # DY995). Quality of BSA is critical (see Technical Hints).

Substrate Solution: 1:1 mixture of Color Reagent A (H2O2) and Color Reagent B (Tetramethylbenzidine) (Catalog # DY999)

Stop Solution: 2 N H2SO4 (Catalog # DY994)

Microplates: R&D Systems (Catalog # DY990), or equivalent

Plate Sealers: ELISA Plate Sealers (Catalog # DY992), or equivalent

 

Data Example

Human Galectin-3 ELISA Standard Curve

Product Datasheets

Preparation and Storage

Shipping
The product is shipped at ambient temperature. Upon receipt, store it immediately at the temperature recommended below.
Stability & Storage
Store the unopened product at 2 - 8 °C. Do not use past expiration date.

Background: Galectin-3

The galectins constitute a large family of carbohydrate-binding proteins that function in many systems both intracellularly and following secretion. Galectins contain either one or two carbohydrate recognition domains (CRR) which mediate recognition of N-acetyl-lactosamine-containing glycoproteins. Some galectins exist in multiple isoforms due to alternative splicing. Individual galectins differ in their tissue distribution and in their carbohydrate-binding specificities.

Entrez Gene IDs:
3958 (Human); 16854 (Mouse)
Alternate Names:
AGE-R3; Carbohydrate-binding protein 35; CBP 35,35 kDa lectin; CBP35; GAL3; Gal-3; galactin-3; Galactose-specific lectin 3; Galactoside-binding protein; GALBPCBP35; Galectin3; Galectin-3; GALIG; IgE-binding protein; L29; L31; L-31; Laminin-binding protein; Lectin L-29; lectin, galactoside-binding, soluble, 3; LGALS2; LGALS3; Mac-2 antigen; Mac-2; MAC2GAL3

Assay Procedure

GENERAL ELISA PROTOCOL

Plate Preparation

  1. Dilute the Capture Antibody (to the working concentration stated in the product datasheet ) in PBS without carrier protein. Immediately coat a 96-well microplate with 100 µL per well of the diluted Capture Antibody. Seal the plate and incubate overnight at room temperature.
  2. Aspirate each well and wash with Wash Buffer, repeating the process two times for a total of three washes. Wash by filling each well with Wash Buffer (400 µL) using a squirt bottle, manifold dispenser, or autowasher. Complete removal of liquid at each step is essential for good performance. After the last wash, remove any remaining Wash Buffer by aspirating or by inverting the plate and blotting it against clean paper towels.
  3. Block each well of the microplate as recommended in the product datasheet. Incubate at room temperature for a minimum of 1 hour.

    Note: The recommended Reagent Diluent typically contains 1% BSA. Some DuoSet Development Kits require alternative blocking agents, or for plates to be blocked overnight with a higher percentage of BSA, please see the product datasheet for details.
     
  4. Repeat the aspiration/wash as in step 2. The plates are now ready for sample addition.

 

PRECAUTION
The Stop Solution suggested for use with this kit is an acid solution. Wear eye, hand, face and clothing protection when using this material.

Assay Procedure

  1. Add 100 µL of sample or standards in Reagent Diluent, or an appropriate diluent, per well. Cover with an adhesive strip and incubate 2 hours at room temperature.
  2. Repeat the aspiration/wash as in step 2 of Plate Preparation.
  3. Add 100 µL of the Detection Antibody, diluted in Reagent Diluent (as recommended in the product datasheet), to each well. Cover with a new adhesive strip and incubate 2 hours at room temperature.
  4. Repeat the aspiration/wash as in step 2 of Plate Preparation.
  5. Add 100 µL of the working dilution of Streptavidin-HRP to each well. Cover the plate and incubate for 20 minutes at room temperature. Avoid placing the plate in direct light.
  6. Repeat the aspiration/wash as in step 2.
  7. Add 100 µL of Substrate Solution to each well. Incubate for 20 minutes at room temperature. Avoid placing the plate in direct light.
  8. Add 50 µL of Stop Solution to each well. Gently tap the plate to ensure thorough mixing.
  9. Determine the optical density of each well immediately, using a microplate reader set to 450 nm. If wavelength correction is available, set to 540 nm or 570 nm. If wavelength correction is not available, subtract readings at 540 nm or 570 nm from the readings at 450 nm. This subtraction will correct for optical imperfections in the plate. Readings made directly at 450 nm without correction may be higher and less accurate.

Citations for Human Galectin-3 DuoSet ELISA

R&D Systems personnel manually curate a database that contains references using R&D Systems products. The data collected includes not only links to publications in PubMed, but also provides information about sample types, species, and experimental conditions.

7 Citations: Showing 1 - 7
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  1. The Phenotype and Secretory Activity of Adipose-Derived Mesenchymal Stem Cells (ASCs) of Patients with Rheumatic Diseases
    Authors: E Kuca-Warna, U Skalska, I Janicka, U Musia?owic, K Bonek, P G?uszko, P Szcz?sny, M Olesi?ska, E Kontny
    Cells, 2019;8(12):.
    Species: Human
    Sample Types: Cell Culture Supernates
  2. Fecal Galectin-3: A New Promising Biomarker for Severity and Progression of Colorectal Carcinoma
    Authors: M Jovanovic, N Gajovic, N Zdravkovic, M Jovanovic, M Jurisevic, D Vojvodic, V Maric, A Arsenijevi, I Jovanovic
    Mediators Inflamm., 2018;2018(0):8031328.
    Species: Human
    Sample Types: Feces
  3. Galectin-3 acts as an angiogenic switch to induce tumor angiogenesis via Jagged-1/Notch activation
    Authors: SN Dos Santos, H Sheldon, JX Pereira, C Paluch, EM Bridges, MC El-Cheikh, AL Harris, ES Bernardes
    Oncotarget, 2017;0(0):.
    Species: Human
    Sample Types: Cell Culture Supernates
  4. Increased expression of colonic Wnt9A through Sp1-mediated transcriptional effects involving arylsulfatase B, chondroitin 4-sulfate, and galectin-3.
    Authors: Bhattacharyya S, Feferman L, Tobacman J
    J Biol Chem, 2014;289(25):17564-75.
    Species: Human
    Sample Types: Cell Lysates
  5. Hypoxia reduces arylsulfatase B activity and silencing arylsulfatase B replicates and mediates the effects of hypoxia.
    Authors: Bhattacharyya S, Tobacman JK
    PLoS ONE, 2012;7(3):e33250.
    Species: Human
    Sample Types: Cell Lysates
  6. Human breast cancer cells enhance self tolerance by promoting evasion from NK cell antitumor immunity.
    Authors: Mamessier E, Sylvain A, Thibult ML, Houvenaeghel G, Jacquemier J, Castellano R, Goncalves A, Andre P, Romagne F, Thibault G, Viens P, Birnbaum D, Bertucci F, Moretta A, Olive D
    J. Clin. Invest., 2011;121(9):3609-22.
    Species: Human
    Sample Types: Cell Culture Supernates
  7. Novel INTeraction of MUC4 and galectin: potential pathobiological implications for metastasis in lethal pancreatic cancer.
    Authors: Senapati S, Chaturvedi P, Chaney WG, Chakraborty S, Gnanapragassam VS, Sasson AR, Batra SK
    Clin. Cancer Res., 2011;17(2):267-74.
    Species: Human
    Sample Types: Serum

FAQs

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Reviews for Human Galectin-3 DuoSet ELISA

Average Rating: 4.5 (Based on 2 Reviews)

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Human Galectin-3 DuoSet ELISA
By Anonymous on 06/13/2019
Application: Sample Tested: EDTA Plasma

Human Galectin-3 DuoSet ELISA
By Anonymous on 05/24/2016
Application: Sample Tested: EDTA Plasma

We run human EDTA plasma samples diluted 1:2 in Reagent Diluent 1x with this kit.