Mouse Adiponectin/Acrp30 DuoSet ELISA

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DY1119
Ancillary Products Available
Adiponectin/Acrp30 ELISA Kit ELISA
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Citations (30)
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Mouse Adiponectin/Acrp30 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 mouse Adiponectin / Acrp30. 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 Adiponectin / Acrp30 ELISA Standard Curve

Product Datasheets

Preparation and Storage

Stability & Storage
Store the unopened product at 2 - 8 °C. Do not use past expiration date.

Background: Adiponectin/Acrp30

Adiponectin (also known as Adipocyte complement-related protein of 30 kDa, Acrp30) is an adipocyte-derived protein with wide ranging paracrine and endocrine effects on metabolism and inflammation. It promotes adipocyte differentiation, fatty acid catabolism, and insulin sensitivity and is negatively correlated with obesity, type 2 diabetes, and atherogenesis. Adiponectin associates into 70 kDa noncovalently-linked trimers that are disulfide-linked into medium molecular weight hexamers and then into >300 kDa high molecular weight oligomers.

Entrez Gene IDs:
9370 (Human); 11450 (Mouse); 246253 (Rat)
Alternate Names:
ACDC; Acrp30; ACRP30ADPN; adipocyte, C1Q and collagen domain containing; Adiponectin; adiponectin, C1Q and collagen domain containing; AdipoQ; ADIPQTL1; ApM1; apM-1; APM1APM-1; C1q and collagen domain-containing protein; GBP28; GBP28apM1; Gelatin-binding protein

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 Adiponectin/Acrp30 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.

30 Citations: Showing 1 - 10
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  1. GPS2 Deficiency Triggers Maladaptive White Adipose Tissue Expansion in Obesity via HIF1A Activation
    Authors: K Drareni, R Ballaire, S Barilla, MJ Mathew, A Toubal, R Fan, N Liang, C Chollet, Z Huang, M Kondili, F Foufelle, A Soprani, R Roussel, JF Gautier, F Alzaid, E Treuter, N Venteclef
    Cell Rep, 2018;24(11):2957-2971.e6.
    Species: Mouse
    Sample Types: Blood
  2. Acute physical exercise increases the adaptor protein APPL1 in the hypothalamus of obese mice
    Authors: RC Gaspar, VR Muñoz, GP Formigari, GK Kuga, SCBR Nakandakar, JD Botezelli, ASR da Silva, DE Cintra, LP de Moura, ER Ropelle, JR Pauli
    Cytokine, 2018;110(0):87-93.
    Species: Mouse
    Sample Types: Serum
  3. Metformin Mitigates Fibrosis and Glucose Intolerance Induced by Doxorubicin in Subcutaneous Adipose Tissue
    Authors: LA Biondo, HA Batatinha, CO Souza, AAS Teixeira, LS Silveira, MI Alonso-Val, LM Oyama, MJ Alves, M Seelaender, JCR Neto
    Front Pharmacol, 2018;9(0):452.
    Species: Mouse
    Sample Types: Serum
  4. Collagen overlays can inhibit leptin and adiponectin secretion but not lipid accumulation in adipocytes
    Authors: SL Christian, NK Pallegar, RJ Brown, AM Viloria-Pe
    PeerJ, 2018;6(0):e4641.
    Species: Mouse
    Sample Types: Cell Culture Supernates
  5. Egg white hydrolysate shows insulin mimetic and sensitizing effects in 3T3-F442A pre-adipocytes
    Authors: F Jahandideh, S Chakrabart, ST Davidge, J Wu
    PLoS ONE, 2017;12(10):e0185653.
    Species: Mouse
    Sample Types: Cell Culture Supernates
  6. 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
  7. Mathematical modeling of white adipocyte exocytosis predicts adiponectin secretion and quantifies the rate of vesicle exo- and endocytosis
    Authors: C Brännmark, W Lövfors, AM Komai, T Axelsson, MF El Hachman, S Musovic, A Paul, E Nyman, CS Olofsson
    J. Biol. Chem., 2017;0(0):.
    Species: Mouse
    Sample Types: Cell Culture Supernates
  8. Obesity-induced diet leads to weight gain, systemic metabolic alterations, adipose tissue inflammation, hepatic steatosis, and oxidative stress in gerbils (Meriones unguiculatus)
    Authors: LL Ventura, NC Fortes, HC Santiago, MV Caliari, MA Gomes, DR Oliveira
    PeerJ, 2017;5(0):e2967.
    Species: Gerbil
    Sample Types: Tissue Homogenates
  9. 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
  10. Mathematical modeling of white adipocyte exocytosis predicts adiponectin secretion and quantifies the rate of vesicle exo- and endocytosis
    Authors: C Brännmark, W Lövfors, AM Komai, T Axelsson, MF El Hachman, S Musovic, A Paul, E Nyman, CS Olofsson
    J. Biol. Chem., 2017;0(0):.
    Species: Mouse
    Sample Types: Cell Culture Supernates
  11. Exosomes isolated from sera of mice fed Lactobacillus strains affect inflammatory cytokine production in macrophages in�vitro
    Authors: A Aoki-Yoshi, S Saito, T Tsuruta, A Ohsumi, H Tsunoda, K Sonoyama
    Biochem. Biophys. Res. Commun., 2017;0(0):.
    Species: Mouse
    Sample Types: Serum
  12. Inducing maternal inflammation promotes leptin production in offspring but does not improve allergic symptoms in a mouse model of allergic rhinitis
    Authors: A Imai, K Satoi, E Fujimoto, K Sato
    Heliyon, 2017;3(6):e00327.
    Species: Mouse
    Sample Types: Serum
  13. Bavachin from Psoralea corylifolia Improves Insulin-Dependent Glucose Uptake through Insulin Signaling and AMPK Activation in 3T3-L1 Adipocytes
    Authors: H Lee, H Li, M Noh, JH Ryu
    Int J Mol Sci, 2016;17(4):.
    Species: Mouse
    Sample Types: Cell Culture Supernates
  14. Hepatic Expression of Adenovirus 36 E4ORF1 Improves Glycemic Control and Promotes Glucose Metabolism via AKT Activation
    Authors: Travis B McMurphy
    Diabetes, 2016;0(0):.
    Species: Mouse
    Sample Types: Serum
  15. Palmitoleic Acid Improves Metabolic Functions in Fatty Liver by PPAR?-Dependent AMPK Activation
    Authors: Camila O Souza
    J. Cell. Physiol, 2016;0(0):.
    Species: Mouse
    Sample Types: Serum
  16. Loss of CD24 in Mice Leads to Metabolic Dysfunctions and a Reduction in White Adipocyte Tissue.
    Authors: Fairbridge N, Southall T, Ayre D, Komatsu Y, Raquet P, Brown R, Randell E, Kovacs C, Christian S
    PLoS ONE, 2015;10(11):e0141966.
    Species: Mouse
    Sample Types: Plasma
  17. Obesity increases airway hyperresponsiveness via the TNF-alpha pathway and treating obesity induces recovery.
    Authors: Kim J, Sohn J, Lee J, Park J
    PLoS ONE, 2015;10(2):e0116540.
    Species: Mouse
    Sample Types: Tissue Homogenates
  18. Milk-derived tripeptides IPP (Ile-Pro-Pro) and VPP (Val-Pro-Pro) promote adipocyte differentiation and inhibit inflammation in 3T3-F442A cells.
    Authors: Chakrabarti S, Wu J
    PLoS ONE, 2015;10(2):e0117492.
    Species: Mouse
    Sample Types: Cell Culture Supernates
  19. The adipokine Retnla modulates cholesterol homeostasis in hyperlipidemic mice.
    Authors: Lee M, Lim C, Lee Y, Park J, Sonn S, Lee M, Jung I, Jeong S, Jeon S, Lee M, Oh K, Yang Y, Kim J, Choi H, Jeong W, Jeong T, Yoon W, Kim H, Choi J, Oh G
    Nat Commun, 2014;5(0):4410.
    Species: Mouse
    Sample Types: Serum
  20. Increased expression of c-Jun in nonalcoholic fatty liver disease.
    Authors: Dorn C, Engelmann J, Saugspier M, Koch A, Hartmann A, Muller M, Spang R, Bosserhoff A, Hellerbrand C
    Lab Invest, 2014;94(4):394-408.
    Species: Mouse
    Sample Types: Serum
  21. Interleukin-15 modulates adipose tissue by altering mitochondrial mass and activity.
    Authors: Barra, Nicole G, Palanivel, Rengasam, Denou, Emmanuel, Chew, Marianne, Gillgrass, Amy, Walker, Tina D, Kong, Josh, Richards, Carl D, Jordana, Manel, Collins, Stephen, Trigatti, Bernardo, Holloway, Alison C, Raha, Sandeep, Steinberg, Gregory, Ashkar, Ali A
    PLoS ONE, 2014;9(12):e114799.
    Species: Mouse
    Sample Types: Cell Culture Supernates
  22. Lactoferrin dampens high-fructose corn syrup-induced hepatic manifestations of the metabolic syndrome in a murine model.
    Authors: Li Y, Hsieh C
    PLoS ONE, 2014;9(5):e97341.
    Species: Mouse
    Sample Types: Tissue Homogenates
  23. 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
  24. Metabolic profile response to administration of epigallocatechin-3-gallate in high-fat-fed mice.
    Authors: Moreno M, De Laquila R, Okuda M, Lira F, de Souza G, de Souza C, Telles M, Ribeiro E, do Nascimento C, Oyama L
    Diabetol Metab Syndr, 2014;6(1):84.
    Species: Mouse
    Sample Types: Tissue Homogenates
  25. 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
  26. Effect of secretin on preadipocyte, differentiating and mature adipocyte functions.
    Authors: Miegueu P, Cianflone K, Richard D, St-Pierre D
    Int J Obes (Lond), 2013;37(3):366-74.
    Species: Mouse
    Sample Types: Cell Culture Supernates
  27. Deficiency of C5L2 increases macrophage infiltration and alters adipose tissue function in mice.
    Authors: Gauvreau D, Gupta A, Fisette A, Tom F, Cianflone K
    PLoS ONE, 2013;8(4):e60795.
    Species: Mouse
    Sample Types: Plasma
  28. Elevated levels of circulating IL-18BP and perturbed regulation of IL-18 in schizophrenia.
    J Neuroinflammation, 2012;9(0):206.
    Species: Human
    Sample Types: Serum
  29. Rosiglitazone improves survival and hastens recovery from pancreatic inflammation in obese mice.
    Authors: Pini M, Rhodes DH, Castellanos KJ
    PLoS ONE, 2012;7(7):e40944.
    Species: Mouse
    Sample Types: Plasma
  30. 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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Mouse Adiponectin/Acrp30 DuoSet ELISA
By Anonymous on 04/26/2018
Sample Tested: Pancreatic cancer cells