Figure 1. PSA/KLK3 protein is expressed in LNCaP cells but absent in other cell lines, as detected using the PathScan® Total PSA/KLK3 Sandwich ELISA Kit #14119. The absorbance readings at 450 nm are shown in the top figure, while the corresponding western blot using PSA/KLK3 (D6B1) XP® Rabbit mAb #5365 is shown in the bottom figure.
Figure 2. The relationship between protein concentration of lysates from LNCaP cells and the absorbance at 450 nm as detected by the PathScan® Total PSA/KLK3 Sandwich ELISA Kit is shown. LNCaP cells (85% confluence) were harvested and lysed.
|Product Includes||Volume||Solution Color|
|PSA/KLK3 Rabbit mAb Coated Microwells||96 tests|
|PSA/KLK3 Mouse Detection mAb||1 ea||Green (Lyophilized)|
|Anti-mouse IgG, HRP-linked Antibody (ELISA Formulated)||1 ea||Red (Lyophilized)|
|Detection Antibody Diluent||11 ml||Green|
|HRP Diluent||11 ml||Red|
|TMB Substrate 7004||11 ml|
|STOP Solution 7002||11 ml|
|Sealing Tape||2 ea|
|ELISA Wash Buffer (20X) 9801||25 ml|
|ELISA Sample Diluent||25 ml||Blue|
|Cell Lysis Buffer (10X) 9803||15 ml|
PathScan® Total PSA/KLK3 Sandwich ELISA Kit is a solid phase sandwich enzyme-linked immunosorbent assay (ELISA) that detects endogenous levels of PSA/KLK3. A PSA/KLK3 Rabbit mAb has been coated onto the microwells. After incubation with cell lysates, PSA/KLK3 protein is captured by the coated antibody. Following extensive washing, a PSA/KLK3 Mouse Detection mAb is added to detect the captured PSA/KLK3 protein. A HRP-linked, anti-mouse antibody is then used to recognize the bound detection antibody. HRP substrate, TMB, is added to develop color. The magnitude of the absorbance for this developed color is proportional to the quantity of total PSA/KLK3.
Antibodies in kit are custom formulations specific to kit.
NOTE: Prepare solutions with purified water.
*NOTE: Some PathScan® ELISA Kits may include HRP-Linked Streptavidin in place of HRP-Linked Antibody.
NOTE: Initial color of positive reaction is blue, which changes to yellow upon addition of STOP Solution.
posted November 2013
Protocol Id: 204
PathScan® Total PSA/KLK3 Sandwich ELISA Kit detects endogenous levels of PSA/KLK3 protein as shown in Figure 1. The kit sensitivity is shown in Figure 2. This kit detects proteins from the indicated species, as determined through in-house testing, but may also detect homologous proteins from other species.Species Reactivity:
Kallikrein 3 (KLK3), also known as Prostate Specific Antigen (PSA), is a member of the glandular kallikrein subfamily of serine proteases (1). It is produced by prostate epithelial cells and is secreted into prostatic ducts. Upon cleavage of 7 amino-terminal amino acids (2), it is activated to liquefy semen in the seminal coagulum. Although PSA/KLK3 is produced in healthy individuals, investigators have found abnormally high levels in the blood of men with advanced prostate cancer (2,3).
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