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hKLK3 QKit (1 Kit)
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hKLK3 QKit (1 Kit)

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品牌: R&D Systems
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Product Summary

The Quantikine Human KLK3/PSA Immunoassay is a 4.5 hour solid phase immunoassay designed to measure KLK3/PSA in cell culture supernates, serum, and plasma. It contains NS0-expressed recombinant human KLK3/PSA, and antibodies raised against the recombinant protein. Natural human KLK3/PSA showed dose-response curves that were parallel to the standard curves obtained using the recombinant Quantikine kit standards, indicating that this kit can be used to determine relative levels of natural human KLK3/PSA.

Recovery

The recovery of KLK3/PSA spiked to levels throughout the range of the assay in various matrices was evaluated.

Sample TypeAverage % RecoveryRange %
Cell Culture Media (n=4)9993-105
EDTA Plasma (n=4)10093-103
Heparin Plasma (n=4)10197-109
Serum (n=4)10297-109

Linearity

To assess the linearity of the assay, samples containing and/or spiked with high concentrations of KLK3/PSA were serially diluted with the appropriate Calibrator Diluent to produce samples with values within the dynamic range of the assay.
Human Kallikrein 3/PSA ELISA Linearity

Scientific Data

Human Kallikrein 3/PSA ELISA Cell Culture Supernate Standard Curve

Human Kallikrein 3/PSA ELISA Serum/Plasma Standard Curve

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 the appropriate 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.
  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.

Human Kallikrein 3/PSA 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)
Sensitivity
0.069 ng/mL
Assay Range
0.9 - 60 ng/mL (Cell Culture Supernates, Serum, EDTA Plasma, Heparin Plasma)
Specificity
Natural and recombinant human KLK3/PSA
Cross-reactivity
< 0.5% cross-reactivity observed with available related molecules.Cross-species reactivity not tested.
Interference
No significant interference observed with available related molecules.
背景
别名
APS seminin,EC 3.4.21,EC 3.4.21.77,Gamma-seminoprotein,hK3,Kallikrein 3,kallikrein 3, (prostate specific antigen),Kallikrein-3,kallikrein-related peptidase 3,KLK2A1,KLK3,P-30 antigen,prostate specific antigen,prostate-specific antigen,PSA semenogelase,PSA,Semenogelase,Seminin
背景

Background: Kallikrein 3/PSA

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:
354 (Human)
Alternate Names:
APS seminin; EC 3.4.21; EC 3.4.21.77; Gamma-seminoprotein; hK3; Kallikrein 3; kallikrein 3, (prostate specific antigen); Kallikrein-3; kallikrein-related peptidase 3; KLK2A1; KLK3; P-30 antigen; prostate specific antigen; prostate-specific antigen; PSA semenogelase; PSA; Semenogelase; Seminin
制备和贮存
保存方式

Preparation and Storage

Shipping
The product is shipped at ambient temperature. Upon receipt, store it immediately at the temperature recommended below.
Storage
Store the unopened product at 2 - 8 °C. Do not use past expiration date.
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货号:
DKK300
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