Figure 1. Androgen receptor protein from human (HeLa, MCF7, T-47D, LNCaP, or U-87 MG) cells was detected using PathScan® Total Androgen Receptor Sandwich ELISA Kit. However, this kit has no detection in human (Jurkat, K-562, or PC-3), monkey (COS-7), hamster (CHO), mouse (NIH/3T3, C2C12), or rat (C6, PC-12) cells. The absorbance readings at 450 nm are shown in the top figure, while corresponding western blots using Androgen Receptor (D6F11) XP® Rabbit mAb #5153 are shown in the bottom figure.
Figure 2. The relationship between protein concentration of lysates from LNCaP cells and the absorbance at 450 nm is shown. LNCaP cells (85% confluence) were harvested and then lysed.
|12850C||1 Kit (96 assays)||$ 513|
|Product Includes||Volume||Solution Color|
|Androgen Receptor Rabbit mAb Coated Microwells||96 tests|
|Androgen Receptor 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|
The PathScan® Total Androgen Receptor Sandwich ELISA Kit is a solid phase sandwich enzyme-linked immunosorbent assay (ELISA) that detects endogenous levels of total androgen receptor protein. An Androgen Receptor Rabbit mAb has been coated onto the microwells. After incubation with cell lysates, androgen receptor protein is captured by the coated antibody. Following extensive washing, an Androgen Receptor Mouse Detection mAb is added to detect the captured androgen receptor protein. Anti-mouse IgG, HRP-linked Antibody is then used to recognize the bound detection antibody. HRP substrate, TMB, is added to develop color. The magnitude of absorbance for the developed color is proportional to the quantity of total androgen receptor protein.
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 Androgen Receptor Sandwich ELISA Kit recognizes endogenous levels of androgen receptor 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.
Androgen receptor (AR), a zinc finger transcription factor belonging to the nuclear receptor superfamily, is activated by phosphorylation and dimerization upon ligand binding (1). This promotes nuclear localization and binding of AR to androgen response elements in androgen target genes. Research studies have shown that AR plays a crucial role in several stages of male development and the progression of prostate cancer (2,3).
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