Mouse/Rat Angiopoietin-2 Quantikine ELISA Kit

R&D Systems | Catalog # MANG20

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Key Product Details

Assay Length

4.5 hours

Sample Type & Volume Required Per Well

Cell Culture Supernates (50 µL), Tissue Lysates (10 µL), Serum (10 µL), EDTA Plasma (10 µL), Heparin Plasma (10 µL)

Sensitivity

4.98 pg/mL

Assay Range

39.1-2500 pg/mL (Cell Culture Supernates, Tissue Lysates, Serum, EDTA Plasma, Heparin Plasma)
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Product Summary for Mouse/Rat Angiopoietin-2 Quantikine ELISA Kit

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.

Product Specifications

Assay Type

Solid Phase Sandwich ELISA

Format

96-well strip plate

Measurement

Quantitative ELISA

Detection Method

Colorimetric - 450nm (TMB)

Conjugate

HRP

Species

Mouse, Rat

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.

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, EDTA Plasma, Heparin Plasma, Serum, Tissue Lysates

Intra-Assay Precision Inter-Assay Precision
Sample 1 2 3 1 2 3
n 20 20 20 20 20 20
Mean (pg/mL) 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 for Mouse/Rat Angiopoietin-2 Quantikine ELISA Kit

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

Scientific Data Images for Mouse/Rat Angiopoietin-2 Quantikine ELISA Kit

Mouse/Rat Angiopoietin-2 ELISA Standard Curve

Mouse/Rat Angiopoietin-2 ELISA Standard Curve

Mouse/Rat Angiopoietin-2 Quantikine ELISA Kit

Mouse/Rat Angiopoietin-2 Quantikine ELISA Kit

Endothelial permeability in response to CLP. Plasma levels of endocan (A) and angiopoietin‐2 (B), soluble markers of endothelial permeability were measured in Sham and CLP rats (n = 8/group) via ELISA. *P < 0.05 versus Sham. Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/31054188), licensed under a CC-BY license. Not internally tested by R&D Systems.

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: Angiopoietin-2

Angiopoietin-2 (Ang-2; also ANGPT2) is a secreted glycoprotein that regulates vascular remodeling by promoting endothelial cell survival, proliferation, and migration and destabilizing the interaction between EC and perivascular cells. Ang-2 is required for postnatal vascular remodeling, and it cooperates with Ang-1 during lymphatic vessel development. It mediates the upregulation of ICAM-1 and VCAM-1 on EC, which facilitates the adhesion of leukocytes during inflammation. Its direct interaction with Integrins promotes tumor cell invasion. Ang-2 is widely expressed during development, but it is restricted postnatally to highly angiogenic tissues such as the placenta, ovaries, and uterus. It associates into dimers, trimers, and larger multimers and binds the receptor Tie-2, with its function as an agonist or antagonist depending on its state of multimerization.

Alternate Names

ANGPT2

Entrez Gene IDs

285 (Human); 11601 (Mouse); 89805 (Rat); 396730 (Porcine); 101925495 (Cynomolgus Monkey)

Gene Symbol

ANGPT2

Additional Angiopoietin-2 Products

Product Documents for Mouse/Rat Angiopoietin-2 Quantikine ELISA Kit

Certificate of Analysis

To download a Certificate of Analysis, please enter a lot or batch number in the search box below.

Note: Certificate of Analysis not available for kit components.

Product Specific Notices for Mouse/Rat Angiopoietin-2 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 Mouse/Rat Angiopoietin-2 Quantikine ELISA Kit

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Protocols

View specific protocols for Mouse/Rat Angiopoietin-2 Quantikine ELISA Kit (MANG20):

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
  3.   Add 50 µL of Assay Diluent to each well.
    4. 50 µL Standard, Control, or Sample
  4.   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.
  5.   Aspirate each well and wash, repeating the process 3 times for a total of 4 washes.
    6. 100 µL Conjugate
  6.   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.
  7.   Aspirate and wash 4 times.
    8. 100 µL Substrate Solution
  8.   Add 100 µL Substrate Solution to each well. Incubate at room temperature for 30 minutes on the benchtop. PROTECT FROM LIGHT.
    9. 100 µL Stop Solution
  9.   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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FAQs

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