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Human RBP4 ELISA Kit - Quantikine

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Human RBP4/Retinol-Binding Protein 4 ELISA Standard Curve
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Summary Sample Values Precision Recovery Linearity Data Examples Product Datasheets Preparation and Storage Background Related Research Areas

Human RBP4 Quantikine ELISA Kit Summary

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.

Sample Values

Serum/Plasma/Saliva/Urine - Samples from apparently healthy volunteers were evaluated for the presence of human RBP4 in this assay. No medical histories were available for the donors used in this study.

Sample TypeMean (pg/mL)Range (pg/mL)Standard Deviation (pg/mL)
Serum (n=35)26,50012,700-48,6007700
EDTA plasma (n=35)25,20011,900-48,3007200
Heparin plasma (n=35)25,00012,200-43,0007500
Urine (n=11)85.216.4-25266.8
Salive (n=12)14.93.11-29.610.0

Cell Culture Supernates:
Human peripheral blood mononuclear cells (1 x 106 cells/mL) were cultured in DMEM supplemented with 5% fetal bovine serum, 5 μM beta -mercaptoethanol, 2 mM L-glutamine, 100 U/mL penicillin, and 100 μg/mL streptomycin sulfate. Cells were cultured unstimulated or stimulated with 10 μg/mL PHA for 1 and 5 days. Aliquots of the cell culture supernates were removed and assayed for levels of human RBP4. No detectable levels were observed. 

HepG2 human hepatocellular carcinoma cells (1 x 106 cells/mL) were cultured in MEM supplemented with non-essential amino acids, 10% fetal bovine serum, and 2 mM L-glutamine for 5 days. An aliquot of the cell culture supernate was removed, assayed for human RBP4, and measured 75.2 ng/mL. 

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
Sample 1 2 3 1 2 3
n 20 20 20 40 40 40
Mean (ng/mL) 11 30.3 61.9 9.8 27.5 54.7
Standard Deviation 0.77 1.73 5.01 0.84 1.6 4
CV% 7 5.7 8.1 8.6 5.8 7.3

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/Retinol-Binding Protein 4 ELISA Linearity

Scientific Data

N/A Human RBP4/Retinol-Binding Protein 4 ELISA Standard Curve View Larger

Human RBP4/Retinol-Binding Protein 4 ELISA Standard Curve

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Preparation and Storage

Shipping
The product is shipped at ambient temperature. Upon receipt, store it immediately at the temperature recommended below.
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
&#9888; WARNING: This product can expose you to chemicals including N,N-Dimethylforamide, which is known to the State of California to cause cancer. For more information, go to www.P65Warnings.ca.gov.

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
  3.   Add 200 µL of Assay Diluent to each well.
    4. 20 µL Standard, Control, or Sample
  4.   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.
  5.   Aspirate each well and wash, repeating the process 3 times for a total of 4 washes.
    6. 200 µL Conjugate
  6.   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.
  7.   Aspirate and wash 4 times.
    8. 200 µL Substrate Solution
  8.   Add 200 µL Substrate Solution to each well. Incubate at room temperature for 30 minutes on the benchtop. PROTECT FROM LIGHT.
    9. 50 µL Stop Solution
  9.   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 for Human RBP4 Quantikine ELISA Kit

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. Association of Vitamin A and D Deficiencies with Infectious Outcomes in Children Undergoing Intensive Induction Therapy for Acute Lymphoblastic Leukemia
    Authors: Penkert, RR;Jones, BG;Tang, L;Su, Y;Jeha, S;Yang, J;Yang, W;Ferrolino, J;Strength, R;Pui, CH;Cross, SJ;Hurwitz, JL;Wolf, J;
    The Journal of pediatrics
    Species: Human
    Sample Types: Serum
  2. Astaxanthin Supplementation Augments the Benefits of CrossFit Workouts on Semaphorin 3C and Other Adipokines in Males with Obesity
    Authors: Supriya, R;Shishvan, SR;Kefayati, M;Abednatanzi, H;Razi, O;Bagheri, R;Escobar, KA;Pashaei, Z;Saeidi, A;Shahrbanian, S;Bagchi, S;Sengupta, P;Al Kiyumi, MH;Heinrich, KM;Zouhal, H;
    Nutrients
    Species: Human
    Sample Types: Plasma
  3. Impaired Organokine Regulation in Non-Diabetic Obese Subjects: Halfway to the Cardiometabolic Danger Zone
    Authors: H L?rincz, B Ratku, S Csiha, I Seres, Z Szabó, G Paragh, M Harangi, S Somodi
    International Journal of Molecular Sciences, 2023-02-18;24(4):.
    Species: Human
    Sample Types: Serum
  4. Temporal changes in the systemic concentrations of retinoids in pregnant and postpartum women
    Authors: H Jeong, AT Armstrong, N Isoherrane, L Czuba, A Yang, K Zumpf, J Ciolino, E Torres, CS Stika, KL Wisner
    PLoS ONE, 2023-02-16;18(2):e0280424.
    Species: Human
    Sample Types: Serum
  5. Vitamin A Status in Preterm Infants Is Associated with Inflammation and Dexamethasone Exposure
    Authors: ME Rossholt, K Wendel, M Bratlie, MF Aas, G Gunnarsdot, D Fugelseth, AH Pripp, M Domellöf, K Størdal, T Stiris, SJ Moltu
    Nutrients, 2023-01-14;15(2):.
    Species: Human
    Sample Types: Whole Blood
  6. Retinol Binding Protein, Sunlight Hours, and the Influenza Virus-Specific Immune Response
    Authors: N Patel, RR Penkert, RE Sealy, SL Surman, BG Jones, K Ringwald-S, AC Ross, JL Hurwitz
    Biomedicines, 2022-09-19;10(9):.
    Species: Human
    Sample Types: Plasma
  7. Hypothesis: Low Vitamin A and D Levels Worsen Clinical Outcomes When Children with Sickle Cell Disease Encounter Parvovirus B19
    Authors: RR Penkert, M Azul, RE Sealy, BG Jones, J Dowdy, RT Hayden, L Tang, AC Ross, JS Hankins, JL Hurwitz
    Nutrients, 2022-08-19;14(16):.
    Species: Human
    Sample Types: Serum
  8. Gut biomolecules (I-FABP, TFF3 and lipocalin-2) are associated with linear growth and biomarkers of environmental enteric dysfunction (EED) in Bangladeshi children
    Authors: MM Hasan, MA Gazi, S Das, SM Fahim, F Hossaini, AR Khan, J Ferdous, MA Alam, M Mahfuz, T Ahmed
    Scientific Reports, 2022-08-16;12(1):13905.
    Species: Human
    Sample Types: Plasma
  9. Association between Maternal Characteristics and Immune Factors TGF-beta1, TGF-beta2, and IgA in Colostrum: An Exploratory Study in Japan
    Authors: N Hirata, M Kiuchi, K Pak, R Fukuda, N Mochimaru, M Mitsui, K Yoshida
    Nutrients, 2022-08-09;14(16):.
    Species: Human
    Sample Types: Breast Milk
  10. Plasma Retinoid Concentrations Are Altered in Pregnant Women
    Authors: LC Czuba, EE Fay, J LaFrance, CK Smith, S Shum, SL Moreni, J Mao, N Isoherrane, MF Hebert
    Nutrients, 2022-03-25;14(7):.
    Species: Human
    Sample Types: Plasma
  11. Posttranslational modifications of proteins are key features in the identification of CSF biomarkers of multiple sclerosis
    Authors: IL Salazar, AST Lourenço, B Manadas, I Baldeiras, C Ferreira, AC Teixeira, VM Mendes, AM Novo, R Machado, S Batista, MDC Macário, M Grãos, L Sousa, MJ Saraiva, AACC Pais, CB Duarte
    Journal of Neuroinflammation, 2022-02-08;19(1):44.
    Species: Human
    Sample Types: Serum
  12. Growth hormone receptor disrupts glucose homeostasis via promoting and stabilizing retinol binding protein 4
    Authors: J Liu, C Nie, L Xue, Y Yan, S Liu, J Sun, M Fan, H Qian, H Ying, L Wang, Y Li
    Theranostics, 2021-07-13;11(17):8283-8300.
    Species: Human
    Sample Types: Serum
  13. The Effects of Beverage Intake after Exhaustive Exercise on Organ Damage, Inflammation and Oxidative Stress in Healthy Males.
    Authors: Tominaga T, Ikemura T, Yada K, Kanda K, Sugama K, Ma S, Choi W, Araya M, Huang J, Nakamura N, Suzuki K
    Antioxidants (Basel), 2021-05-28;10(6):.
    Species: Human
    Sample Types: Urine
  14. Determination of FABP4, RBP4 and the MMP-9/NGAL complex in the serum of women with breast cancer
    Authors: D Tsakogiann, E Kalogera, F Zagouri, E Zografos, D Balalis, G Bletsa
    Oncology Letters, 2020-12-04;21(2):85.
    Species: Human
    Sample Types: Serum
  15. Changes of serum retinol-binding protein 4 associated with improved insulin resistance after laparoscopic sleeve gastrectomy in Chinese obese patients
    Authors: X Wang, Y Huang, J Gao, H Sun, M Jayachandr, S Qu
    Diabetol Metab Syndr, 2020-01-14;12(0):7.
    Species: Human
    Sample Types: Serum
  16. Plasma Levels of Retinol Binding Protein 4 Relate to Large VLDL and Small LDL Particles in Subjects with and without Type 2 Diabetes
    Authors: H Wessel, A Saeed, J Heegsma, MA Connelly, KN Faber, RPF Dullaart
    J Clin Med, 2019-10-25;8(11):.
    Species: Human
    Sample Types: Plasma
  17. The role of renal biomarkers to predict the need of surgery in congenital urinary tract obstruction in infants
    Authors: D Kostic, GPNS Beozzo, SB do Couto, AHT Kato, L Lima, P Palmeira, VLJ Krebs, V Bunduki, RPV Francisco, M Zugaib, FT Dénes, WB de Carvalh, VHK Koch
    J Pediatr Urol, 2019-03-16;0(0):.
    Species: Human
    Sample Types: Urine
  18. Evaluation of serum retinol-binding protein-4 levels as a biomarker of poor short-term prognosis in ischemic stroke
    Authors: YY Zhu, JL Zhang, L Liu, Y Han, X Ge, S Zhao
    Biosci. Rep., 2018-09-12;0(0):.
    Species: Human
    Sample Types: Serum
  19. Elevated RBP4 plasma levels were associated with diabetic retinopathy in type 2 diabetes
    Authors: JY Li, XX Chen, XH Lu, CB Zhang, QP Shi, L Feng
    Biosci. Rep., 2018-09-12;0(0):.
    Species: Human
    Sample Types: Plasma
  20. 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., 2017-08-01;58(10):4095-4103.
    Species: Mouse
    Sample Types: Serum
  21. 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, 2017-07-06;0(0):.
    Species: Human
    Sample Types: Serum
  22. 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, 2017-06-21;2017(0):3937842.
    Species: Human
    Sample Types: Plasma
  23. Low Retinol-Binding Protein and Vitamin D Levels Are Associated with Severe Outcomes in Children Hospitalized with Lower Respiratory Tract Infection and Respiratory Syncytial Virus or Human Metapneumovirus Detection
    Authors: JL Hurwitz, BG Jones, RR Penkert, S Gansebom, Y Sun, L Tang, AM Bramley, S Jain, JA McCullers, SR Arnold
    J. Pediatr., 2017-05-31;0(0):.
    Species: Human
    Sample Types: Serum
  24. 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., 2017-03-31;23(0):185-197.
    Species: Mouse
    Sample Types: Serum
  25. 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, 2015-02-11;13(0):30.
    Species: Human
    Sample Types: Serum
  26. 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, 2012-08-31;11(0):103.
    Species: Human
    Sample Types: Plasma
  27. 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., 2011-10-01;165(4):837-44.
    Species: Human
    Sample Types: Serum
  28. Kinetics of circulating Th17 cytokines and adipokines in psoriasis patients.
    Authors: Nakajima H, Nakajima K, Tarutani M, Morishige R, Sano S
    Arch. Dermatol. Res., 2011-06-17;303(6):451-5.
    Species: Human
    Sample Types: Serum
  29. 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., 2010-02-11;30(5):1043-50.
    Species: Human
    Sample Types: Plasma
  30. Retinol-binding protein-4 in women with untreated essential hypertension.
    Authors: Solini A, Santini E, Madec S, Rossi C, Muscelli E
    Am. J. Hypertens., 2009-06-25;22(9):1001-6.
    Species: Human
    Sample Types: Serum

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