Mouse/Rat Angiopoietin-2 Quantikine ELISA Kit

Catalog # Availability Size / Price Qty
MANG20
Mouse/Rat Angiopoietin-2 Standard Curve
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Mouse/Rat Angiopoietin-2 Quantikine ELISA Kit Summary

Assay Type
Solid Phase Sandwich ELISA
Format
96-well strip plate
Assay Length
4.5 hours
Sample Type & Volume Required Per Well
Cell Culture Supernates (50 uL), Tissue Lysates (10 uL), Serum (10 uL), EDTA Plasma (10 uL), Heparin Plasma (10 uL)
Sensitivity
4.98 pg/mL
Assay Range
39.1 - 2,500 pg/mL (Cell Culture Supernates, Tissue Lysates, Serum, EDTA Plasma, Heparin Plasma)
Specificity
Natural and recombinant mouse and rat Angiopoietin-2.
Cross-reactivity
< 0.5% cross-reactivity observed with available related molecules.< 50% cross-species reactivity observed with species tested.
Interference
No significant interference observed with available related molecules.

Product Summary

The Quantikine Mouse/Rat Angiopoietin-2 Immunoassay is a 4.5 hour solid phase ELISA designed to measure mouse and rat Angiopoietin-2 in cell culture supernates, tissue lysates, serum, and plasma. It contains CHO cell-expressed recombinant mouse Angiopoietin-2 and antibodies raised against the recombinant factor. This immunoassay has been shown to quantitate the recombinant factor accurately. Results obtained using natural Angiopoietin-2 showed dose-response curves that were parallel to the standard curves obtained using the recombinant kit standards. These results indicate that this kit can be used to determine relative mass values for natural Angiopoietin-2.

Precision

Intra-Assay Precision (Precision within an assay) Three samples of known concentration were tested twenty times on one plate to assess intra-assay precision
Inter-Assay Precision (Precision between assays) Three samples of known concentration were tested in twenty separate assays to assess inter-assay precision. Assays were performed by at least three technicians using two lots of kit components

Cell Culture Supernates, Tissue Lysates, Serum, EDTA Plasma, Heparin Plasma

Intra-Assay Precision Inter-Assay Precision
Sample 1 2 3 1 2 3
n 20 20 20 20 20 20
Mean 119 290 831 116 283 841
Standard Deviation 3.27 9.05 22.2 9.84 23.2 63.1
CV% 2.7 3.1 2.7 8.5 8.2 7.5

Recovery

The recovery of Angiopoietin-2 spiked to levels throughout the range of the assay in various matrices was evaluated.

Sample Type Average % Recovery Range %
Cell Culture Samples (n=4) 114 108-120
EDTA Plasma (n=2) 105 101-110
Heparin Plasma (n=2) 107 103-111
Serum (n=2) 113 109-117

Linearity

To assess the linearity of the assay, samples containing and/or spiked with high concentrations of Angiopoietin-2 were serially diluted with Calibrator Diluent to produce samples with values within the dynamic range of the assay.
Mouse/Rat Angiopoietin-2 ELISA Linearity

Product Datasheets

Preparation and Storage

Storage
Store the unopened product at 2 - 8 °C. Do not use past expiration date.

Background: Angiopoietin-2

Angiopoietins, Ang-1, Ang-2, and Ang-3 (mouse)/Ang-4 (human), are natural agonists or antagonists of the Tie-2 receptor tyrosine kinase and are important modulators of angiogenesis. Two domains characterize the angiopoietin family of proteins: an N-terminal coiled-coil domain that mediates homo-oligomerization, and a C-terminal fibrinogen-like domain that binds Tie-2.

Entrez Gene IDs:
285 (Human); 11601 (Mouse)
Alternate Names:
AGPT2; ANG2; ANG-2; angiopoietin 2; Angiopoietin-2; angiopoietin-2a; angiopoietin-2B; angiopoitin 2; ANGPT2; Tie2-ligand
⚠ 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. 50 µL Assay Diluent
  4.   Add 50 µL of Assay Diluent to each well.

  5. 50 µL Standard, Control, or Sample
  6.   Add 50 µL of Standard, Control, or sample to each well. Cover with a plate sealer, and incubate at room temperature for 2 hours on a horizontal orbital microplate shaker.
  7.   Aspirate each well and wash, repeating the process 3 times for a total of 4 washes.

  8. 100 µL Conjugate
  9.   Add 100 µL of Conjugate to each well. Cover with a new plate sealer, and incubate at room temperature for 2 hours on the shaker.
  10.   Aspirate and wash 4 times.

  11. 100 µL Substrate Solution
  12.   Add 100 µL Substrate Solution to each well. Incubate at room temperature for 30 minutes on the benchtop. PROTECT FROM LIGHT.

  13. 100 µL Stop Solution
  14.   Add 100 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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