Human Kallikrein 5 Quantikine ELISA Kit

Catalog # Availability Size / Price Qty
DKK500
Control Products Available
Human Kallikrein 5 Standard Curve
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Product Details
Procedure
Citations (1)
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Human Kallikrein 5 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), Serum (50 uL), EDTA Plasma (50 uL), Heparin Plasma (50 uL), Saliva (13 uL), Human Milk (13 uL)
Sensitivity
30 pg/mL
Assay Range
78.1 - 5,000 pg/mL (Cell Culture Supernates, Serum, EDTA Plasma, Heparin Plasma, Saliva, Human Milk)
Specificity
Natural and recombinant human KLK5
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 Human KLK5 Immunoassay is a 4.5 hour solid-phase ELISA designed to measure human KLK5 in cell culture supernates, serum, plasma, saliva, and human milk. It contains NS0-expressed recombinant human KLK5 and has been shown to accurately quantitate the recombinant factor. Results obtained using natural human KLK5 showed linear curves that were parallel to the standard curves obtained using the Quantikine kit standards. These results indicate that this kit can be used to determine relative mass values for naturally occurring human KLK5.

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 known concentration were tested in separate assays to assess inter-assay precision

Cell Culture Supernates, Serum, EDTA Plasma, Heparin Plasma, Saliva, Human Milk

Intra-Assay Precision Inter-Assay Precision
Sample 1 2 3 1 2 3
n 20 20 20 20 20 20
Mean 510 1685 3407 577 1941 3852
Standard Deviation 35.8 57.2 75.4 63.6 161 278
CV% 7 3.4 2.2 11 8.3 7.2

Recovery

The recovery of KLK5 spiked to levels throughout the range of the assay in various matrices was evaluated.

Sample Type Average % Recovery Range %
Cell Culture Media (n=4) 97 91-109
EDTA Plasma (n=4) 90 82-95
Heparin Plasma (n=4) 91 86-103
Serum (n=4) 93 85-106

Linearity

To assess the linearity of the assay, samples containing and/or spiked with high concentrations of KLK5 were serially diluted with Calibrator Diluent to produce samples with values within the dynamic range of the assay.
Human Kallikrein 5 ELISA Linearity

Data Examples

Human Kallikrein 5 Standard Curve

Product Datasheets

Preparation and Storage

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

Background: Kallikrein 5

The human tissue kallikrein (KLK) gene family contains 15 members that play important roles in cancer. Notably, kallikrein-1, also known as tissue kallikrein, cleaves kininogen to release the vasoactive kinin peptide, bradykinin or lysyl-bradykinin. Kallikrein-3, called prostate specific antigen (PSA), is an established tumor marker that aids in the diagnosis, staging, and follow up of prostate cancer. Kallikrein-4 is specifically expressed in the prostate and over-expressed in prostate cancer. Kallikrein-5 is widely expressed but found at high levels in skin, breast, brain and testis; over-expression is an indicator of poor prognosis in ovarian cancer. Kallikrein-8 is expressed in the brain and is a novel marker of ovarian and cervical cancer.

Human plasma kallikrein, a serine protease, is synthesized in the liver and circulates in the plasma bound to high molecular weight (HMW) kininogen or as a free zymogen. Once activated by its physiological activator, coagulation factor XII, it displays endopeptidase activity towards peptide bonds after arginine (preferred) and lysine. It cleaves HMW kininogen, its major physiological substrate, to release the potent vasodilator peptide bradykinin. It is also able to cleave a number of inactive precursor proteins to generate active products, such as plasminogen and prourokinase.

Entrez Gene IDs:
25818 (Human); 68668 (Mouse); 690603 (Rat)
Alternate Names:
EC 3.4.21; EC 3.4.21.4; Kallikrein 5; Kallikrein-like protein 2; kallikrein-related peptidase 5; KLK5; KLKL2; KLK-L2; KLK-L2EC 3.4.21.-; SCTE; SCTEkallikrein-5; Stratum corneum tryptic enzyme
⚠ 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. 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. Addition of samples must be completed within 15 minutes. Cover with a plate sealer, and incubate at room temperature for 2 hours.
  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.
  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. 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.

Citation for Human Kallikrein 5 Quantikine ELISA Kit

R&D Systems personnel manually curate a database that contains references using R&D Systems products. The data collected includes not only links to publications in PubMed, but also provides information about sample types, species, and experimental conditions.

1 Citation: Showing 1 - 1

  1. Topical Treatment of Rosacea with Ivermectin Inhibits Gene Expression of Cathelicidin Innate Immune Mediators, LL-37 and KLK5, in Reconstructed and Ex Vivo Skin Models
    Authors: S Thibaut de, C Rosignoli, E Soares, M Roquet, B Bertino, JP Chappuis, C Defoin-Pla, D Piwnica
    Dermatol Ther (Heidelb), 2017;0(0):.
    Species: Human
    Sample Types: Cell Culture Supernates

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