Mouse Resistin DuoSet ELISA

Catalog # Availability Size / Price Qty
DY1069
Ancillary Products Available
Mouse Resistin ELISA Standard Curve
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Product Details
Procedure
Citations (6)
FAQs
Supplemental Products
Reviews

Mouse Resistin DuoSet ELISA Summary

Assay Type
Solid Phase Sandwich ELISA
Format
96-well strip plate
Assay Length
4 hours 40 minutes (after plate preparation)
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 mouse Resistin. 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

Mouse Resistin 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: Resistin

Resistin, also known ADSF and FIZZ3, is a secreted peptide hormone that circulates as either a noncovalent trimer, or a disulfide-linked homohexamer. Resistin is expressed by white adipocytes, splenocytes, astrocytes, and anterior pituitary epithelium. It blocks Insulin-stimulated uptake of glucose by adipocytes and promotes glucose release by hepatocytes. High circulating Insulin levels induce Resistin secretion which then antagonize Insulin action.

Entrez Gene IDs:
56729 (Human); 57264 (Mouse)
Alternate Names:
Adipose tissue-specific secretory factor; ADSF; ADSFMGC126609; C/EBP-epsilon regulated myeloid-specific secreted cysteine-rich proteinprecursor 1; C/EBP-epsilon-regulated myeloid-specific secreted cysteine-rich protein; Cysteine-rich secreted protein A12-alpha-like 2; Cysteine-rich secreted protein FIZZ3; FIZZ3; FIZZ3MGC126603; found in inflammatory zone 3; HXCP1; Resistin; RETN; RETN1; RSTNXCP1

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 Mouse Resistin 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.

6 Citations: Showing 1 - 6
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  1. Adiponectin release and insulin receptor targeting share trans-Golgi-dependent endosomal trafficking routes
    Authors: M Rödiger, MW Werno, I Wilhelmi, C Baumeier, D Hesse, N Wettschure, S Offermanns, K Song, M Krau beta, A Schürmann
    Mol Metab, 2017;0(0):.
    Species: Mouse
    Sample Types: Tissue Homogenates
  2. Adipose tissue, metabolic and inflammatory responses to stroke are altered in obese mice
    Authors: MJ Haley, G Mullard, KA Hollywood, GJ Cooper, WB Dunn, CB Lawrence
    Dis Model Mech, 2017;0(0):.
    Species: Mouse
    Sample Types: Tissue Homogenates
  3. Lack of platelet-activating factor receptor protects mice against diet-induced adipose inflammation and insulin-resistance despite fat pad expansion.
    Authors: Menezes-Garcia Z, Oliveira M, Lima R, Soriani F, Cisalpino D, Botion L, Teixeira M, Souza D, Ferreira A
    Obesity (Silver Spring), 2014;22(3):663-72.
    Species: Mouse
    Sample Types: Serum
  4. Hypothalamic gene transfer of BDNF inhibits breast cancer progression and metastasis in middle age obese mice.
    Authors: Liu, Xianglan, McMurphy, Travis, Xiao, Run, Slater, Andrew, Huang, Wei, Cao, Lei
    Mol Ther, 2014;22(7):1275-84.
    Species: Mouse
    Sample Types: Serum
  5. A2B adenosine receptors prevent insulin resistance by inhibiting adipose tissue inflammation via maintaining alternative macrophage activation.
    Authors: Csoka B, Koscso B, Toro G, Kokai E, Virag L, Nemeth Z, Pacher P, Bai P, Hasko G
    Diabetes, 2014;63(3):850-66.
    Species: Mouse
    Sample Types: Plasma
  6. Mutant amyloid precursor protein differentially alters adipose biology under obesogenic and non-obesogenic conditions.
    Authors: Freeman, Linnea R, Zhang, Le, Dasuri, Kalavath, Fernandez-Kim, Sun-Ok, Bruce-Keller, Annadora, Keller, Jeffrey
    PLoS ONE, 2012;7(8):e43193.
    Species: Mouse
    Sample Types: Serum

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