Human copeptin ELISA kit

Instructions
Code CSB-E12130h
Size 96T,5×96T,10×96T
Trial Size 24T ELISA kits trial application
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Product Details

Description

The copeptin ELISA kit is used to detect and quantify protein levels of endogenous copeptin. The assay recognizes human copeptin in serum, urine, cell culture supernates, and tissue homogenates. It provides a detection range of 78 pg/mL-5000 pg/mL. It has a high sensitivity, as low as 19.5 pg/mL. In addition, this kit also possesses high precision, good lineaity, excellent specificity, and high recovery.

This experiment uses the sandwich ELISA technique. The pre-coated antibody on the micro ELISA plate is an anti-human copeptin antibody and the detection antibody is a biotin-conjugated polyclonal copeptin antibody. Samples and biotin labeling antibody are added into wells and thoroughly washed out. Avidin-HRP conjugates are added to wells. The TMB substrate is added for coloring after thorough washing. The solution turns blue in HRP catalytic and finally turns yellow under the action of the stop solution. The color depth is positively correlated to the testing analyte in samples.

Copeptin, the C-terminal part of precursor pre-provasopressin (pre-proAVP), is usually elevated in acute stress and cardiac emergencies. It is secreted together with vasopressin (AVP) equally as a response of AVP stimulation. Therefore, it is a sensitive and stable surrogate marker for the release of arginine AVP. AVP is a key stress hormone involved in hemodynamics and osmoregulation.

Target Name copeptin
Abbreviation copeptin
Species Homo sapiens (Human)
Sample Types serum, urine, cell culture supernates, tissue homogenates
Detection Range 78 pg/mL-5000 pg/mL
Sensitivity 19.5 pg/mL
Assay Time 1-5h
Sample Volume 50-100ul
Detection Wavelength 450 nm
Research Area Others
Assay Principle quantitative
Measurement Sandwich
Precision
Intra-assay Precision (Precision within an assay): CV%<8%
Three samples of known concentration were tested twenty times on one plate to assess.
Inter-assay Precision (Precision between assays): CV%<10%
Three samples of known concentration were tested in twenty assays to assess.
Linearity
To assess the linearity of the assay, samples were spiked with high concentrations of human Copeptin in various matrices and diluted with the Sample Diluent to produce samples with values within the dynamic range of the assay.
 SampleSerum(n=4)
1:1Average %89
Range %82-95
1:2Average %99
Range %92-105
1:4Average %97
Range %90-106
1:8Average %96
Range %91-102
Recovery
The recovery of human Copeptin spiked to levels throughout the range of the assay in various matrices was evaluated. Samples were diluted prior to assay as directed in the Sample Preparation section.
Sample TypeAverage % RecoveryRange
Serum (n=5) 9388-98
Typical Data
These standard curves are provided for demonstration only. A standard curve should be generated for each set of samples assayed.
pg/mlOD1OD2AverageCorrected
50002.535 2.752 2.644 2.533
25002.012 2.179 2.096 1.985
12501.598 1.599 1.599 1.488
6250.847 0.867 0.857 0.746
3120.478 0.502 0.490 0.379
1560.336 0.356 0.346 0.235
780.237 0.221 0.229 0.118
00.109 0.112 0.111  
Materials provided
  • A micro ELISA plate ---The 96-well plate has been pre-coated with an anti-Human copeptin antibody.
  • Two vials lyophilized standard ---Dilute a bottle of the standard at dilution series, read the OD values, and then draw a standard curve.
  • One vial Biotin-labeled copeptin antibody (100 x concentrate) (120 μl/bottle) ---Act as the detection antibody.
  • One vial HRP-avidin (100 x concentrate) (120 μl/bottle) ---Bind to the detection antibody and react with the TMB substrate to make the solution chromogenic.
  • One vial Biotin-antibody Diluent (15 ml/bottle) ---Dilute the Biotin-antibody.
  • One vial HRP-avidin Diluent (15 ml/bottle) ---Dilute the HRP-avidin solution.
  • One vial Sample Diluent (50 ml/bottle) ---Dilute the sample to an appropriate concentration.
  • One vial Wash Buffer (25 x concentrate) (20 ml/bottle) ---Wash away unbound or free substances.
  • One vial TMB Substrate (10 ml/bottle) ---Act as the chromogenic agent. TMB interacts with HRP, eliciting the solution turns blue.
  • One vial Stop Solution (10 ml/bottle) ---Stop the color reaction. The solution color immediately turns from blue to yellow.
  • Four Adhesive Strips (For 96 wells) ---Cover the microplate when incubating.
  • An instruction manual
Materials not provided
  • A microplate reader capable of measuring absorbance at 450 nm, with the correction wavelength set at 540 nm or 570 nm.
  • An incubator can provide stable incubation conditions up to 37°C±5°
  • Centrifuge
  • Vortex
  • Squirt bottle, manifold dispenser, or automated microplate washer
  • Absorbent paper for blotting the microtiter plate
  • 50-300ul multi-channel micropipette
  • Pipette tips
  • Single-channel micropipette with different ranges
  • 100ml and 500ml graduated cylinders
  • Deionized or distilled water
  • Timer
  • Test tubes for dilution
Troubleshooting
and FAQs
ELISA kit FAQs
Storage Store at 2-8°C. Please refer to protocol.
Lead Time 3-5 working days

Citations

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 Q&A
Q:

Please provide the following information regarding human copeptin ELISA Kit (CSB-E12130h)
Please confirm the concentration of the stock? (5000pg/ml)

A:
Thanks for your inquiry.
The standard in the kit is lyophilized powder. The concentration after reconstitution by sample diluent is 5000pg/ml.Pls let me know if you have any further questions. Thank you.

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