Human HTRA2/Omi Quantikine ELISA Kit

    
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  • 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), Serum (25 uL), Platelet-poor EDTA Plasma (25 uL), Platelet-poor Heparin Plasma (25 uL)
  • Sensitivity
    13.2 pg/mL
  • Assay Range
    31.3 - 2,000 pg/mL (Cell Culture Supernates, Serum, Platelet-poor EDTA Plasma, Platelet-poor Heparin Plasma)
  • Specificity
    Natural and recombinant HTRA2/Omi
  • 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.
Control Available
QC103 , Quantikine Immunoassay Control Set 792 for Human HTRA2/Omi - Please Inquire
Product Summary
The Quantikine Human HTRA2/Omi Immunoassay is a 4.5 hour solid phase ELISA designed to measure HTRA2 levels in cell culture supernates, serum, and platelet-poor plasma. It contains E. coil-expressed recombinant human HTRA2 and antibodies raised against the recombinant protein. Results obtained for naturally occurring human HTRA2 and recombinant human HTRA2 showed linear curves that were parallel to the standard curves obtained using the Quantikine Human HTRA2/Omi Immunoassay standards. These results indicate that this kit can be used to determine relative mass values for natural human HTRA2.

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 a 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, Serum, Platelet-poor EDTA Plasma, Platelet-poor Heparin Plasma
Intra-Assay Precision Inter-Assay Precision
Sample 1 2 3 1 2 3
n 20 20 20 20 20 20
Mean 284 582 1108 280 567 1089
Standard Deviation 15.7 15.9 28.6 25.1 41.3 65.3
CV% 5.5 2.7 2.6 9 7.3 6

Recovery

The recovery of human VEGF spiked to levels throughout the range of the assay in various matrices was evaluated.

Sample Type Average % Recovery Range %
Cell Culture Media (n=4) 92 86-101
Platelet-poor EDTA Plasma (n=4) 90 79-100
Platelet-poor Heparin Plasma (n=4) 89 81-95
Serum (n=4) 91 80-111
Linearity
To assess the linearity of the assay, samples containing and/or spiked with high concentrations of HTRA2 were serially diluted with Calibrator Diluent to produce samples with values within the dynamic range of the assay.
 HTRA2/Omi [HRP]
Product Datasheets

Certificate of Analysis Lookup

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Note: Certificate of Analysis not available for kit components.

Certificate of Analysis Lookup

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Note: Certificate of Analysis not available for kit components.

Preparation and Storage
  • Storage
    Store the unopened product at 2 - 8 °C. Do not use past expiration date.
Background: HTRA2/Omi
HTRA2/Omi is the mammalian homolog of the bacterial high temperature requirement protein (HTRA). HTRA2/Omi is a homotrimeric serine protease that localizes to the mitochondria and is processed to expose an N-terminal reaper-like motif similar to SMAC/Diablo. HTRA2/Omi is released from the mitochondria in response to apoptotic insult and can interact with the BIR2 or BIR3 domains of XIAP to relieve caspase-IAP inhibition.
  • Long Name:
    High Temperature Requirement Protein-2
  • Entrez Gene IDs:
    27429 (Human); 64704 (Mouse)
  • Alternate Names:
    High temperature requirement protein A2; HtrA serine peptidase 2; HTRA2; HtrA-like serine protease; Omi; OMIOmi stress-regulated endoprotease; PARK13EC 3.4.21.108; PRSS25; PRSS25serine, 25; Serine protease 25; serine protease HTRA2, mitochondrial; Serine proteinase OMI
Related Research Areas
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. 100 µL Assay Diluent
  4.   Add 100 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. 200 µL Conjugate
  9.   Add 200 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. 200 µL Substrate Solution
  12.   Add 200 µL Substrate Solution to each well. Incubate at room temperature for 30 minutes on the benchtop. PROTECT FROM LIGHT.

  13. 50 µL Stop Solution
  14.   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.

FAQs

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ELISA Controls
Description Application Cat# Citations Images  

Quantikine Immunoassay Control Set 792 for Human HTRA2/Omi

Ctrl QC103
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Supplemental ELISA Products
Description Application Cat# Citations Images  

Quantikine ELISA Wash Buffer 1

ELISA WA126 6  
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