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Human RBP4 Quantikine ELISA Kit

R&D Systems | Catalog # DRB400

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Key Product Details

Assay Length

2.5 hours

Sample Type & Volume Required Per Well

Cell Culture Supernates (20 µL), Serum (10 µL), EDTA Plasma (10 µL), Heparin Plasma (10 µL), Saliva (10 µL), Urine (10 µL)

Sensitivity

0.628 ng/mL

Assay Range

1.56-100 ng/mL (Cell Culture Supernates, Serum, EDTA Plasma, Heparin Plasma, Saliva, Urine)
View all RBP4/Retinol-Binding Protein 4 ELISA Kits »

Product Summary for Human RBP4 Quantikine ELISA Kit

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.

Product Specifications

Assay Type

Solid Phase Sandwich ELISA

Format

96-well strip plate

Measurement

Quantitative ELISA

Detection Method

Colorimetric - 450nm (TMB)

Conjugate

HRP

Species

Human

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. 

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, EDTA Plasma, Heparin Plasma, Saliva, Serum, Urine

Intra-Assay Precision Inter-Assay Precision
Sample 1 2 3 1 2 3
n 20 20 20 40 40 40
Mean (ng/mL) 11.0 30.3 61.9 9.80 27.5 54.7
Standard Deviation 0.77 1.73 5.01 0.84 1.60 4.00
CV% 7.0 5.7 8.1 8.6 5.8 7.3

Recovery for Human RBP4 Quantikine ELISA Kit

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 Images for Human RBP4 Quantikine ELISA Kit

Human RBP4/Retinol-Binding Protein 4 ELISA Standard Curve

Human RBP4/Retinol-Binding Protein 4 ELISA Standard Curve

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: 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.

Alternate Names

RetinolBinding Protein 4

Entrez Gene IDs

5950 (Human); 19662 (Mouse); 25703 (Rat)

Gene Symbol

RBP4

Additional RBP4/Retinol-Binding Protein 4 Products

Product Documents for Human RBP4 Quantikine ELISA Kit

Certificate of Analysis

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Note: Certificate of Analysis not available for kit components.

Product Specific Notices for Human RBP4 Quantikine ELISA Kit

For research use only

⚠ 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.

Citations for Human RBP4 Quantikine ELISA Kit

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Protocols

View specific protocols for Human RBP4 Quantikine ELISA Kit (DRB400):

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.

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