Human RBP4 Quantikine ELISA Kit

(10 citations)   
  • Assay Type
    Solid Phase Sandwich ELISA
  • Format
    96-well strip plate
  • Assay Length
    2.5 hours
  • Sample Type & Volume Required Per Well
    Cell Culture Supernates (20 uL), Serum (10 uL), EDTA Plasma (10 uL), Heparin Plasma (10 uL), Saliva (10 uL), Urine (10 uL)
  • Sensitivity
    0.628 ng/mL
  • Assay Range
    1.6 - 100 ng/mL (Cell Culture Supernates, Serum, EDTA Plasma, Heparin Plasma, Saliva, Urine)
  • Specificity
    Natural and recombinant human RBP4
  • Cross-reactivity
    < 0.5% cross-reactivity observed with available related molecules.Cross-species reactivity not tested.
  • Interference
    No significant interference observed with available related molecules.
Control Available
QC23, Quantikine Immunoassay Control Group 8 - Please Inquire
Product Summary
The Quantikine Human RBP4 immunoassay is a 2.5 hour solid phase ELISA designed to measure human RBP4 in cell culture supernates, serum, plasma, urine, and saliva. It contains NS0-expressed recombinant human RBP4 and has been shown to accurately quantitate the recombinant factor. Results obtained using natural human RBP4 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 RBP4.

Precision
Intra-Assay Precision (Precision within an assay) Three samples of known concentration were tested on one plate to assess intra-assay precision.
Inter-Assay Precision (Precision between assays) Three samples of known concentration were tested in separate assays to assess inter-assay precision.
Cell Culture Supernates, Serum, EDTA Plasma, Heparin Plasma, Saliva, Urine
Intra-Assay Precision Inter-Assay Precision
Sample123123
n202020404040
Mean1130.361.99.827.554.7
Standard Deviation0.771.735.010.841.64
CV%75.78.18.65.87.3

Recovery

The recovery of RBP4 spiked to levels throughout the range of the assay was evaluated.

Sample Type Average % Recovery Range %
Cell Culture Media (n=4) 102 86-115
Linearity
To assess the linearity of the assay, samples containing and/or spiked with high concentrations of RBP4 were serially diluted with the Calibrator Diluent to produce samples with values within the dynamic range of the assay.
Human RBP4 Quantikine ELISA Kit
Preparation and Storage
  • Storage
    Store the unopened product at 2 - 8 °C. Do not use past expiration date.
Background: RBP4/Retinol-Binding Protein 4
Retinol-binding protein 4 (RBP4) is a Lipocalin superfamily molecule that transports vitamin A (retinol) and retinaldehyde in the serum. Retinaldehyde is the critical chromophore in the rhodopsin photoreceptor, while both it and retinoic acid regulate a multitude of cellular differentiation and proliferation effects through the intracellular receptors RAR and RXR. RBP4 is secreted primarily by hepatocytes and adipocytes into the blood where the RBP4-retinol complex interacts with transthyretin (TTR). Formation of the complex with TTR increases the serum half-life of RBP4 by preventing RBP4 filtration through the kidney. The C-terminally processed forms of RBP4, which do not bind TTR, are normally excreted into the urine but accumulate in the serum during renal failure. RBP4 promotes hyperglycemia, and its upregulation in visceral and liver adipocytes leads to elevated serum levels.
    • Entrez Gene IDs
      5950 (Human); 19662 (Mouse); 25703 (Rat);
    • Alternate Names
      interstitial; Plasma retinol-binding protein; RBP4; retinol binding protein 4, plasma; RetinolBinding Protein 4; retinol-binding protein 4, plasma;
    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. 200 µL Assay Diluent
    4.   Add 200 µL of Assay Diluent to each well.

    5. 20 µL Standard, Control, or Sample
    6.   Add 20 µL of Standard, control, or sample to each well. Cover with a plate sealer, and incubate at room temperature for 1 hour on a horizontal orbital microplate shaker.
    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 1 hour on the shaker.
    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 on the benchtop. 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.

    10 Citations: Showing 1 - 10
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    Species
    Sample Type
    1. Serum retinol-binding protein-induced endothelial inflammation is mediated through the activation of toll-like receptor 4
      Authors: M Du, A Martin, F Hays, J Johnson, RA Farjo, KM Farjo
      Mol. Vis., 2017;23(0):185-197.
      Species: Mouse
      Sample Type: Serum
    2. Transgenic Mice Over-Expressing RBP4 Have RBP4-Dependent and Light-Independent Retinal Degeneration
      Authors: M Du, E Phelps, MJ Balangue, A Dockins, G Moiseyev, Y Shin, S Kane, L Otalora, JX Ma, R Farjo, KM Farjo
      Invest. Ophthalmol. Vis. Sci., 2017;58(10):4095-4103.
      Species: Mouse
      Sample Type: Serum
    3. RBP4-STRA6 Pathway Drives Cancer Stem Cell Maintenance and Mediates High-Fat Diet-Induced Colon Carcinogenesis
      Authors: S Karunanith, L Levi, J DeVecchio, G Karagkouni, O Reizes, JD Lathia, MF Kalady, N Noy
      Stem Cell Reports, 2017;0(0):.
      Species: Human
      Sample Type: Serum
    4. Concurrent Aerobic and Resistance Training Has Anti-Inflammatory Effects and Increases Both Plasma and Leukocyte Levels of IGF-1 in Late Middle-Aged Type 2 Diabetic Patients
      Authors: G Annibalini, F Lucertini, D Agostini, L Vallorani, A Gioacchini, E Barbieri, M Guescini, L Casadei, A Passalia, M Del Sal, G Piccoli, M Andreani, A Federici, V Stocchi
      Oxid Med Cell Longev, 2017;2017(0):3937842.
      Species: Human
      Sample Type: Plasma
    5. Proxy markers of serum retinol concentration, used alone and in combination, to assess population vitamin A status in Kenyan children: a cross-sectional study.
      Authors: Talsma E, Verhoef H, Brouwer I, Mburu-de Wagt A, Hulshof P, Melse-Boonstra A
      BMC Med, 2015;13(0):30.
      Species: Human
      Sample Type: Serum
    6. Adipocytokine levels mark endothelial function in normotensive individuals.
      Authors: Solini A, Stea F, Santini E, Bruno R, Duranti E, Taddei S, Ghiadoni L
      Cardiovasc Diabetol, 2012;11(0):103.
      Species: Human
      Sample Type: Plasma
    7. Increased retinol-binding protein (RBP) 4 and anti-RBP4 antibody in alopecia areata.
      Authors: Ahn KJ, Choi EA, Kim J, Lee JH, Lee KH, Bang D, Cho SB
      Br. J. Dermatol., 2011;165(4):837-44.
      Species: Human
      Sample Type: Serum
    8. Kinetics of circulating Th17 cytokines and adipokines in psoriasis patients.
      Authors: Nakajima H, Nakajima K, Tarutani M, Morishige R, Sano S
      Arch. Dermatol. Res., 2011;303(6):451-5.
      Species: Human
      Sample Type: Serum
    9. Nonalcoholic fatty liver disease as the transducer of hepatic oversecretion of very-low-density lipoprotein-apolipoprotein B-100 in obesity.
      Authors: Chan DC, Watts GF, Gan S
      Arterioscler. Thromb. Vasc. Biol., 2010;30(5):1043-50.
      Species: Human
      Sample Type: Plasma
    10. Retinol-binding protein-4 in women with untreated essential hypertension.
      Authors: Solini A, Santini E, Madec S, Rossi C, Muscelli E
      Am. J. Hypertens., 2009;22(9):1001-6.
      Species: Human
      Sample Type: Serum
    ELISA Controls
    Description Application Cat# Citations Images  

    Quantikine Immunoassay Control Group 8

    Ctrl QC23
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    Supplemental ELISA Products
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    Quantikine ELISA Wash Buffer 1

    ELISA WA126 6
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