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Kallikrein-2 ELISA Kit (KLK2)

Full Name: Kallikrein-2 ELISA Kit (KLK2)
Reactivity: Human
Sample Type: Serum, Tissue Homogenates, Plasma, Biological Fluids
Sensitivity: 0.094 ng/ml

INTRODUCTION

Kallikrein-2 (KLK2) is a protein-coding gene, located on chromosome 19q13.33. Primarily expressed in the prostate gland and plays a crucial role in prostate physiology and pathology. A single-chain glycosylated serine protease enzyme that consists of 244 amino acids. It has a molecular weight of 28 kDa. The crystal structure reveals a typical serine protease fold, with a central beta-sheet surrounded by alpha-helices. The active site of KLK2 contains a serine residue that is essential for catalytic activity. Structurally it is classified as an alpha/beta/epsilon-type serine protease with eight beta-strands and four epsilon-helices. The beta-sheets are surrounded by eight alpha helices, making the shape of the molecule overall a long and thin one. The active site has been found to contain a serine residue that is essential for catalytic activity in humans, but not in mouse or rat counterparts.

In the physiology and pathophysiology of the prostate, kallikrein-2 is an important player. PSA is processed through proteolysis by KLK2, which is expressed in the epithelial cells of the prostate gland. It transforms the inactive pro-PSA into the active form of PSA by cleaving it. The correct measurement of PSA levels in blood, which are frequently employed for the identification and monitoring of prostate cancer, depends on this conversion. Kallikrein-2 is also over-expressed in prostate cancer, leading to a lack of PSA cleavage. It facilitates cancer cell invasion by inhibiting the expression of EphA2 and PTEN, as well as inhibiting the motility of normal cells.

The membrane-bound form of kallikrein-2 is typically about 15 kDa, which is replaced by a 26–28 kDa intracellular form that is phosphorylated. The plasma membrane bound form has a molecular weight of about 2250 kDa, with subunits that are approximately 30kD each. In contrast, the nuclear DNA protein translated from mRNA takes on a molecular weight of 2480–2510 kDa, with subunits ranging from 27–39kD.

INTENDED USE

Human kallikrein-2 ELISA kit is designed for analysing amounts of KLK2 (kallikrein-2, glandular kallikrein-1, hGK, KLK-2) using human serum, plasma, tissue homogenates and many other biological fluids.

CONTENT

All reagents supplied need to be stored at 2 °C – 8 °C, unopened reagents will retain reactivity until expiration date. Do not use reagents beyond this date.

  • One 96-Well Plate: Pre-coated with anti KLK2 antibody.
  • Standards: Lyophilized recombinant.
  • Sample/Standard Dilution Buffer.
  • Biotinylated-labelled Antibody.
  • Antibody Dilution Buffer.
  • HRP-Streptavidin Conjugate (SABC).
  • SABC Dilution Buffer.
  • TMB Substrate.
  • Wash Buffer (25x).
  • Plate Sealer.
  • Product Instructions.

TYPICAL RESULTS

For this kallikrein-2 ELISA kit it is recommended that a standard curve is generated for each assay carried out.

Standard Curve: 0, 0.156, 0.312, 0.625, 1.25, 5, 10 ng/ml.
Reactivity: Human
Sensitivity: 0.094 ng/ml
Range: 0.156 – 10 ng/ml
Principle: Sandwich
Application: Research Use Only.

ASSAY CHARACTERISTICS

– Specificity: Highly specific for KLK2, no cross reactivity or interference between KLK2 and analogues was detected.
– Recovery: Serum (88 – 103%), EDTA Plasma (91 – 103%), Heparin Plasma (85 – 104%).
– Linearity: Serum (85 – 96%), EDTA Plasma (86 – 100%), Heparin Plasma (87 – 97%).
– Precison Intra-Assay: CV < 8%.
– Precison Inter-Assay: CV < 10%.
– Stability: Less than 10%.

REFERENCES

  1. Prostate specific antigen and human glandular kallikrein 2 in early detection of prostate cancer. J Urol. (2003) 169 (2): 445-57. Karazanashvili G. and Abrahamsson P.A.
  2. Targeting kallikrein-related peptidases in prostate cancer. Expert Opin Ther Targets. (2014) 18 (4): 365-83. Mavridis K, et al.
  3. Unleashing the therapeutic potential of human kallikrein-related serine proteases. Nat Rev Drug Discov. (2015) 14 (3): 183-202. Prassas I., et al.
  4. Human kallikrein-2-gene polymorphism is associated with the occurrence of prostate cancer. J Urol. (2005) 173 (2): 429-32. Chiang C.H., et al.

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