|46484C||1 Kit (96 assays)||
|Product Includes||Volume||Solution Color|
|Claudin-6 Rabbit mAb Coated Microwells||96 assays|
|Claudin-6 Rabbit Detection mAb||1 ea||Red (Lyophilized)|
|HRP Diluent||5.5 ml||Red|
|TMB Substrate 7004||11 ml|
|STOP Solution 7002||11 ml|
|Sealing Tape||2 ea|
|ELISA Wash Buffer (20X) 9801||25 ml|
|Cell Lysis Buffer (10X) 9803||15 ml|
The rapid protocol (RP) PathScan® RP Claudin-6 Sandwich ELISA Kit is a solid phase sandwich enzyme-linked immunosorbent assay (ELISA) that detects endogenous levels of Claudin-6 protein in a reduced assay time of 1.5 hours. Incubation of cell lysates and detection antibody on the coated microwell plate forms a sandwich with Claudin-6 protein in a single step. The plate is then extensively washed and TMB reagent is added for signal development. The magnitude of absorbance for the developed color is proportional to the quantity of Claudin-6 protein. Learn more about your ELISA kit options here.
*Antibodies in this kit are custom formulations specific to kit.
NOTE: This protocol is for PathScan® kits that use an HRP directly conjugated to the detection antibody (Rapid Protocol), rather than a 2-step method where the detection antibody and a secondary-HRP are added sequentially.
NOTE: Prepare solutions with deionized/purified water or equivalent.
For adherent cells
For suspension cells
NOTE: Equilibrate all materials and prepared reagents to room temperature prior to running the assay.
NOTE: Initial color of positive reaction is blue, which changes to yellow upon addition of STOP Solution.
created July 2020
Protocol Id: 2144
Tight junctions, or zonula occludens, form a continuous barrier to fluids across the epithelium and endothelium. They function in regulation of paracellular permeability and in the maintenance of cell polarity, blocking the movement of transmembrane proteins between the apical and the basolateral cell surfaces. Tight junctions are composed of claudin and occludin proteins, which join the junctions to the cytoskeleton (1,2). The claudin family is composed of 23 integral membrane proteins, and their expression, which varies among tissue types, may determine both the strength and properties of the epithelial barrier. Alteration in claudin protein expression pattern is associated with several types of cancer (2,3). Claudin-1 is expressed primarily in keratinocytes (4) and normal mammary epithelial cells, but is absent or reduced in breast carcinomas and breast cancer cell lines (5,6).
Claudin-6 is a member of the CLDN family that is expressed in epithelial cell sheets. Downregulation of Claudin-6 has been reported in breast invasive ductal carcinoma associated with lymphatic metastasis, which may point to a function of Claudin-6 as a tumor suppressor. Claudin-6 is reported to play a role in inhibiting malignancy of breast cancer cells by inducing apoptosis, inhibiting proliferation and migration. Mechanisms of action of Claudin-6 have been described through various signaling pathways such as p38-MAPK, JAKs-STATs, ASK1-JNK, and other pathways (7,8). Regulation of Claudin-6 expression may occur through epigenetic mechanisms (9). Other reports describe aberrant expression in various malignancies (10,11). The clinical significance of Claudin-6 dysregulation has created interest in the potential for pharmaceutical intervention (12-14).
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