Human FABP4 Quantikine ELISA Kit

  (2 citations)     
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Assay Procedure
Citations (2)
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  • Assay Type
    Solid Phase Sandwich ELISA
  • Format
    96-well strip plate
  • Assay Length
    4.5 hours
  • Sample Type & Volume Required Per Well
    Cell Culture Supernates (50 uL), Cell Lysates (50 uL), Serum (10 uL), EDTA Plasma (10 uL), Heparin Plasma (10 uL)
  • Sensitivity
    14.2 pg/mL
  • Assay Range
    62.5 - 4,000 pg/mL (Cell Culture Supernates, Cell Lysates, Serum, EDTA Plasma, Heparin Plasma)
  • Specificity
    Natural and recombinant human FABP4.
  • Cross-reactivity
    < 0.5% cross-reactivity observed with available related molecules.< 50% cross-species reactivity observed with species tested.
  • Interference
    No significant interference observed with available related molecules.
Product Summary
The Quantikine Human FABP4 Immunoassay is a 4.5 hour solid-phase ELISA designed to measure human FABP4 in cell culture supernates, cell lysates, serum, and plasma. It contains E. coli-expressed recombinant human FABP4 and has been shown to accurately quantitate the recombinant factor. Results obtained using natural human FABP4 showed linear curves that were parallel to the standard curves obtained using the Quantikine kit standards. These results indicate that this kit can be used to determine relative mass values for naturally occurring human FABP4.

Precision
Intra-Assay Precision (Precision within an assay) Three samples of known concentration were tested twenty times on one plate to assess intra-assay precision
Inter-Assay Precision (Precision between assays) Three samples of known concentration were tested in twenty separate assays to assess inter-assay precision. Assays were performed by at least three technicians using two lots of components
Cell Culture Supernates, Cell Lysates, Serum, EDTA Plasma, Heparin Plasma
Intra-Assay Precision Inter-Assay Precision
Sample 1 2 3 1 2 3
n 20 20 20 20 20 20
Mean 349 1155 2595 407 1236 2619
Standard Deviation 20.2 39.7 117 51.6 156 291
CV% 5.8 3.4 4.5 12.7 12.6 11.1

Recovery

The recovery of human FABP4 spiked to levels throughout the range of the assay in various matrices was evaluated.

Sample Type Average % Recovery Range %
Cell Culture Media (n=4) 108 100-118
Cell Lysis Buffer 1 (n=2) 108 92-117
EDTA Plasma (n=4) 98 88-113
Heparin Plasma (n=4) 95 83-103
Serum (n=4) 99 82-113
Linearity
To assess the linearity of the assay, samples containing and/or spiked with high concentrations of human FABP4 were diluted with Calibrator Diluent to produce samples with values within the dynamic range of the assay.
 FABP4/A-FABP [HRP]
Product Datasheets

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Preparation and Storage
  • Storage
    Store the unopened product at 2 - 8 °C. Do not use past expiration date.
Background: FABP4/A-FABP
FABP4 (Fatty Acid Binding Protein 4), also known as A-FABP and aP2, is the predominant FABP found in adipocytes and is often used as a marker for adipocyte differentiation. It is also expressed in macrophages, dendritic cells, and endothelial cells. FABP4 is a key mediator of intracellular fatty acid transport and metabolism in adipose tissue. Its expression is regulated by multiple factors including fatty acids, PPAR gamma agonists, and Insulin, and its levels increase with lipolytic stimulation. It can increase the hydrolytic activity of Hormone-Sensitive Lipase and increase the production of proinflammatory cytokines and chemokines. FABP4 upregulation in adipocytes and macrophages is associated with the development of Insulin resistance, hypertriacylglycerolaemia, and atherosclerosis. Serum levels of FABP4 are elevated in obesity and metabolic syndrome, type 2 diabetes, HIV-associated lipodystrophy, polycystic ovary syndrome, nonalcoholic fatty liver disease, atherosclerosis, and acute ischaemic stroke. FABP4 is also overexpressed in multiple cancer types including ovarian, bladder, glioblastoma, and oral, and it may play a role in tumor progression.
  • Long Name:
    Fatty Acid-Binding Protein 4
  • Entrez Gene IDs:
    2167 (Human); 11770 (Mouse); 79451 (Rat)
  • Alternate Names:
    Adipocyte lipid-binding protein; Adipocyte-type fatty acid-binding protein; AFABP; A-FABP; A-FABPAFABP; ALBP; aP2; FABP4; fatty acid binding protein 4, adipocyte; Fatty acid-binding protein 4; fatty acid-binding protein, adipocyte
Related Research Areas
Assay Procedure
Refer to the product for complete assay procedure.

Bring all reagents and samples to room temperature before use. It is recommended that all samples, standards, and controls be assayed in duplicate.
  1.   Prepare all reagents, standard dilutions, and samples as directed in the product insert.
  2.   Remove excess microplate strips from the plate frame, return them to the foil pouch containing the desiccant pack, and reseal.

  3. 100 µL Assay Diluent
  4.   Add 100 µL of Assay Diluent to each well.

  5. 50 µL Standard, Control, or Sample
  6.   Add 50 µL of Standard, control, or sample to each well. Cover with a plate sealer, and incubate at room temperature for 2 hours.
  7.   Aspirate each well and wash, repeating the process 3 times for a total of 4 washes.

  8. 200 µL Conjugate
  9.   Add 200 µL of Conjugate to each well. Cover with a new plate sealer, and incubate at room temperature for 2 hours.
  10.   Aspirate and wash 4 times.

  11. 200 µL Substrate Solution
  12.   Add 200 µL Substrate Solution to each well. Incubate at room temperature for 30 minutes. PROTECT FROM LIGHT.

  13. 50 µL Stop Solution
  14.   Add 50 µL of Stop Solution to each well. Read at 450 nm within 30 minutes. Set wavelength correction to 540 nm or 570 nm.
Citations:

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.

2 Citations: Showing 1 - 2
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Species
Sample Type
  1. Adipocyte Fatty-Acid Binding Protein is Overexpressed in Cirrhosis and Correlates with Clinical Outcomes
    Authors: I Graupera, M Coll, E Pose, C Elia, S Piano, E Solà, D Blaya, P Huelin, C Solé, R Moreira, G de Prada, N Fabrellas, A Juanola, M Morales-Ru, P Sancho-Bru, C Villanueva, P Ginès
    Sci Rep, 2017;7(1):1829.
    Species: Human
    Sample Type: Plasma
  2. Using Neonatal Skin to Study the Developmental Programming of Aging
    Authors: Leryn J Reynolds
    Exp. Gerontol, 2016;0(0):.
    Species: Human
    Sample Type: Cell Lysates

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