Figure 1. Detection of CD31 (PECAM-1) protein in various cells is examined by using the PathScan® Total CD31 (PECAM-1) Sandwich ELISA Kit. The absorbance readings at 450 nm are shown in the top figure, while the corresponding
Western blot using CD31 (PECAM-1) (89C2) Mouse mAb #3528 is shown in the bottom figure.
Figure 2. The relationship between protein concentration of lysates from HUVEC cells and the absorbance at 450 nm as detected by the PathScan® Total CD31 (PECAM-1) Sandwich ELISA Kit is shown. Unstarved HUVEC cells were harvested and then lysed.
|Product Includes||Volume||Solution Color|
|CD31 Mouse mAb Coated Microwells||96 tests|
|CD31 (PECAM-1) Biotinylated Detection mAb||1 ea||Green (Lyophilized)|
|HRP-Linked Streptavidin (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 CD31 (PECAM-1) Sandwich ELISA Kit is a solid phase sandwich enzyme-linked immunosorbent assay (ELISA) that detects endogenous levels of CD31 (PECAM-1) protein. A CD31 (PECAM-1) mouse mAb has been coated onto the microwells. After incubation with cell lysates, the CD31 (PECAM-1) proteins are captured by the coated antibody. Following extensive washing, a CD31 (PECAM-1) biotinylated detection mAb is added to detect captured CD31 (PECAM-1) proteins. HRP-linked Streptavidin 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 CD31 (PECAM-1) protein.
Antibodies in the kit are custom formulations specific to the 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
revised January 2016
Protocol Id: 205
PathScan® Total CD31 (PECAM-1) Sandwich ELISA Kit detects endogenous levels of CD31 (PECAM-1) protein in human cells, 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.
CD31 (Platelet Endothelial Cell Adhesion Molecule-1: PECAM-1), a member of the Ig superfamily of cell adhesion molecules, is expressed by circulating platelets, monocytes, neutrophils, some T cells, and endothelial cells and modulates cell adhesion, endothelial cell migration, and angiogenesis (1). CD31 is phosphorylated on Tyr686 at the cytoplasmic carboxy-terminal tail upon various stimuli (e.g. mechanical or oxidative stress), presumably by Src family members (2). The tyrosine phosphorylation mediates associations with a number of SH2 domain-containing binding partners such as PI3 kinase, SHIP, PLCγ, and SHP-2. Thus, CD31 serves as a scaffold for various signaling molecules (3).
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