PathScan Sandwich ELISA Kits
PathScan® Phospho-DDR1 (panTyr) Sandwich ELISA Kit #7863
| Item # | Description | List Price | Web Price | QTY | Buy |
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| Applications | Reactivity | Sensitivity | MW (kDa) | Source |
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| H |
Applications Key:
Reactivity Key:
H = Human
Species cross-reactivity is determined by Western blot.
Specificity / Sensitivity
PathScan® Phospho-DDR1 (panTyr) Sandwich ELISA Kit detects endogenous levels of tyrosine-phosphorylated DDR1 protein in human cells, as shown in Figure 1. The kit sensitivity is shown in Figure 2.
Description
The PathScan® Phospho-DDR1 (panTyr) Sandwich ELISA Kit is a solid phase sandwich enzyme-linked immunosorbent assay (ELISA) that detects endogenous levels of tyrosine-phosphorylated DDR1 protein. A DDR1 rabbit antibody has been coated on the microwells. After incubation with cell lysates, DDR1 protein (phospho and nonphospho) is captured by the coated antibody. Following extensive washing, a phospho-tyrosine mouse mAb is added to detect captured tyrosine-phosphorylated DDR1 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 the absorbance for this developed color is proportional to the quantity of DDR1 protein phosphorylated on tyrosine residues.
Sensitivity
Figure 2: The relationship between protein concentration of phospho or nonphospho lysates and the absorbance at 450 nm is shown. Calu-3 cells were cultured (85% confluence) and lysed with or without the addition of phosphatase inhibitor to the lysis buffer (phospho or nonphospho lysate, respectively).
ELISA - Western correlation
Figure 1: Constitutive phosphorylation of DDR1 in Calu-3 cells lysed in the presence of phosphatase inhibitors* (phospho lysate) is detected by PathScan® Phospho-DDR1 (panTyr) Sandwich ELISA Kit #7863 (upper, right). In contrast, a low level of phospho-DDR1 protein is detected in Calu-3 cells lysed in the absence of phosphatase inhibitors* (nonphospho lysate). Similar levels of total DDR1 protein from both nonphospho and phospho lysates are detected by PathScan® Total DDR1 Sandwich ELISA Kit #7845 (upper, left). Absorbance at 450 nm is shown in the top figure while corresponding western blots using a Phospho-DDR1 (Tyr792/796/797) rabbit antibody (right) or a total DDR1 rabbit antibody (left) are shown in the bottom figure. *Phosphatase inhibitors include sodium pyrophosphate, β-glycerophosphate and Na3VO4.
Background
The discoidin domain receptors (DDRs) are receptor tyrosine kinases with a discoidin homology repeat in their extracellular domains, activated by binding to extracellular matrix collagens. So far, two mammalian DDRs have been identified: DDR1 and DDR2 (1). They are widely expressed in human tissues and may have roles in smooth muscle cell-mediated collagen remodeling (2). Aberrant expression and signaling of DDRs have been implicated in human disease related to increased matrix degradation and remodeling such as cardiovascular disease, liver fibrosis and tumor invasion (1).
- Vogel, W. et al. (1999) FASEB J.13 Suppl, S77 - S82.
- Ferri, N. et al. (2004) Am. J. Pathol.164, 1575 - 1585.
Application References
Have you published research involving the use of our products? If so we'd love to hear about it. Please let us know!
Companion Products
- 7845 PathScan® Total DDR1 Sandwich ELISA Kit
- 3917 DDR1 Antibody
- 7002 STOP Solution
- 9803 Cell Lysis Buffer (10X)
- 9998 BSA
- 9809 Phosphate Buffered Saline with Tween 20 (PBST-20X)
- 7076 Anti-mouse IgG, HRP-linked Antibody
- 9808 Phosphate Buffered Saline (PBS-20X)
- 7004 TMB Substrate
This product is for in vitro research use only and is not intended for use in humans or animals. This product is not intended for use as therapeutic or in diagnostic procedures.
