Call 408.493.1800 | Fax 408.493.1801 | Toll Free 800.891.9699 (US Only) | Email: [email protected]

RBP4 (human) Competitive ELISA Kit

No cross-reactivity with mouse/rat RBP4
Catalog #: K4911

In stock

$835.00

Product Details

Cat # +Size K4911-100
Size 100 assays
Detection Method Absorbance (450 nm)
Species Reactivity Mammalian
Applications This assay detects human RBP4 in the range of 0.001 – 5 µg RBP4/ml and a detection limit of 1 ng/ml (based on adding two standard deviations to the mean value of the (50) zero standards).
Features & Benefits • Simple procedure
• Fast and convenient
• High-throughput adaptable
• Cross Reactivity: No significant cross-reactivity or interference between this analyte and its analogues was observed.
Kit Components • Pre-coated Microtiter Plate
• Wash Buffer (10X)
• Diluent (5X)
• Detection Antibody
• Detector 100X (Hrp conjugated anti-IgG)
• Mouse RBP4 Standard (lyophilized, 24 ng)
• Mouse RBP4 QC Sample (lyophilized)
• Substrate Solution I (TMB)
• Substrate Solution II (Peroxidase)
• Stop Solution
• Plate Sealers
Storage Conditions 4°C
Shipping Conditions Gel Pack
USAGE For Research Use Only! Not For Use in Humans.

Details

Retinol binding protein (RBP) 4 is the only specific transport protein for vitamin A in the circulation whose function is to deliver vitamin to target tissues. In obesity and type 2 diabetes, the expression of Glut4 is significantly impaired in adipocytes. Glucose transport via Glut4 is the rate-limiting step for glucose use by muscle and adipose tissue. Adipocyte-specific deletion of Gluts leads to notable elevation of mouse RBP4 causing systemic insulin resistance, and that reduction of RBP4 improves insulin resistance. This identified a novel role of RBP4 in regulating insulin action and RBP4 is recorded as an adipocyte-derived hormone. The RBP4 (human) ELISA Kit is to be used for the in vitro quantitative determination of human RBP4 in serum, urine and cell culture supernatant. This assay is a competitive ELISA which utilizies a 96-well microtiter plate which was pre-coated with a human RBP4. A purified polyclonal which recognizing native human RBP4 reacts with a series of predetermined recombinant human RBP4 standard proteins or the test samples under competition in the human RBP4-coated plate. Their relative reactivity is plotted with that of the standard proteins. This ELISA is specific for the measurement of natural and recombinant human RBP4. It does not cross-react with mouse RBP4, rat RBP4, human adiponectin, rat adiponectin, human resistin, human vaspin, human clusterin, human leptin, human IL-23, human IL-33, human GPX3, human Nampt, human ANG1, human ANG2, human ANGPTL3, human ANGPTL4, human ANGPTL6, human FABP4, human RELM-β, rat RELM-α, mouse Nampt, human PAI-1. The assay range is 0.001 – 5 µg RBP4/ml and a detection limit of 1 ng/ml (based on adding two standard deviations to the mean value of the (50) zero standards).


Why buy BioVision Products?
Innovation
Affordability
Global Presence
Technical Support
BioVision aims to provide our customers innovative tools for accelerating drug discovery and biological research. BioVision offers >8,000 products including the most comprehensive array of assay kits for key targets in Metabolic pathways.
BioVision is committed to providing the highest quality products at a competitive price.
We have a broad network of global distributors who are ready to address your research needs and ensure fast delivery.
Our highly trained Technical Support team provides comprehensive product support and is dedicated to resolving your issues quickly and efficiently.
What is a sandwich ELISA?
The analyte of interest in a sample is "sandwiched" by an immobilized capture antibody coated on the plate and a labeled antibody used for detection. Sandwich ELISA is very sensitive. The density of color is proportional to the amount of analyte captured from the samples and can be quantified when compared with standard curve.
What is a competitive ELISA?
The endogenous unlabled analyte of interest in a sample is "competed" by the exogenous labeled antigen coated on the plate for a limited amount of antibody binding sites. Therefore, the lower signal indicate higher concentration of the analyte.
What dilution range should I use for the samples?
A preliminary experiment is always recommended when working with new samples and ELISA Kits to ensure all results fall within the detection range.
Can I extend the standard curve (in either direction)?
Extended standard curve is not recommended by BioVision.
What type of software is needed to graph a 4-parameter or 5-parameter curve?
SoftMax Pro by Molecular Devices, SigmaPlot® by Systat Software Inc., or others can be used for this purpose.
Low absorbance; No signal
Target present below detection limits of assay
Reduce dilution factor to increase sample concentration  (Preliminary experiment is recommended for optimal dilution range)
Incompatible sample types
Use samples recommended by the kit
Standard dilutions are unstable
Use fresh standard dilutions
Buffers/substrates are contaminated
Use new buffers and substrates
Insufficient incubation time 
Extend incubation time
Cover or seal plates during all incubation 
Plate washings too vigorous
Pipette wash buffer gently or make sure the automatic wash system has correct pressure
Wells scratched with pipette or washing tips
Use caution when dispensing and aspirating into and out of wells
Incubation temperature too low
Ensure incubations are carried out at temperatures recommended by the manual
Reagents are cold
Bring all reagents to room temperature before each assay
Plate read at incorrect wavelength

Ensure the plate reader is set at the correct wavelength recommended on the protocol

High Background
Insufficient washing
Increase number of washes; Add a 30 second soaking step in between washes;  at the end of each washing step, invert plate on absorbent tissue to completely drain all wells
Residual buffer/substrates remain in wells
Remove all residual buffer and substrates at the end of each wash
Too much HRP-Streptavidin
Centrifuge and mix the substrate vial before use to avoid percipitation and inconsistent HRP concentration
Excessive incubation time
Reduce incubation time
Substrate incubation carry out in light or wait too long to read plate after adding stop solution
Avoid light during incubation and read signals immediately after adding stop solution
Standards improperly reconstituted or diluted
Briefly spin vials before opening; inspect for undissolved material after reconstitution
Poor standard curve
Standard improperly diluted
Confirm dilutions are made correctly
Standard dilutions are unstable
Use freshly diluted standards
Pipetting error 
use calibrated pipette and stay consistent between each pipette
Curve doesn't fit scale
Try plotting using different scales  (log-log, 4 parameter logistic, 5 parameter etc.)
Large intra-coefficient of variation (CV)
Bubbles in wells
Avoid bubbles during experiment, eliminate all bubbles prior to reading
Insufficient washing
Increase number of washes; add a 30 second soak step in between washes;  at the end of each washing step, invert plate on absorbent tissue to completely drain all wells
Wells not washed equally/thoroughly
Use calibrated (automated) multi-channel pipette for consistent handling, make sure all parts of the automated washer are unobstructed
Edge effects
Seal the plate completely during incubation
Incomplete reagent mixing
Make sure all reagents are mixed thoroughly during each step
Inconsistent sample preparation or storage
Consistent sample preparation. Optimize sample storage conditions and minimize freeze and thaw cycles
Stacked plates
Avoid stacking plates during incubation
Plate sealers not used or reused
Cover assay plate with plate sealer, use a fresh sealer each time to prevent contamination
Large inter-coefficient of variation (CV))
Insufficient washing
Increase number of washes; add a 30 second soak step in between washes;  at the end of each washing step, invert plate on absorbent tissue to completely drain all wells
Inconsistent incubation temperature
Make sure to follow recommended incubation temperatures. Avoid fluctuations in temperature due to environmental conditions.
Plate sealers not used or reused
Cover assay plate with plate sealer, use a fresh sealer each time to prevent contamination
$
Subscribe to our Newsletter!!
and automatically get enrolled into a raffle to win Google Home.
Offer valid for US customers only.