Product Pathways - PathScan ELISA
PathScan® Total DDR1 Sandwich ELISA Kit #7845
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|Kit Includes||Volume||Solution Color|
|DDR1 Rabbit Antibody Coated Microwells||96 tests|
|DDR1 Mouse Detection mAb||11 ml||Green|
|Anti-mouse IgG, HRP-linked Antibody||11 ml||Red|
|TMB Substrate #7004||11 ml||Colorless|
|STOP Solution #7002||11 ml||Colorless|
|Sealing Tape||2 sheets|
|ELISA Wash Buffer (20X)||25 ml||Colorless|
|Sample Diluent||25 ml||Blue|
|Cell Lysis Buffer (10X) #9803||15 ml||Yellowish|
Note: 12 8-well modules –Each module is designed to break apart for 8 tests.
Note: Kit should be stored at 4°C with the exception of Cell Lysis Buffer, which is stored at –20°C (packaged separately).
Species enclosed in parentheses are predicted to react based on 100% sequence homology.
The PathScan® Total DDR1 Sandwich ELISA Kit is a solid phase sandwich enzyme-linked immunosorbent assay (ELISA) that detects endogenous levels of total 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 DDR1 mouse mAb is added to detect captured 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 total DDR1 protein.
Specificity / Sensitivity
PathScan® Total DDR1 Sandwich ELISA Kit #7845 detects endogenous levels of total DDR1 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.
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.
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).
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). Research studies have implicated aberrant expression and signaling of DDRs in human diseases related to increased matrix degradation and remodeling, such as cardiovascular disease, liver fibrosis, and tumor invasion (1).
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- 7863 PathScan® Phospho-DDR1 (panTyr) Sandwich ELISA Kit
- 5583 DDR1 (D1G6) XP® Rabbit mAb
- 3917 DDR1 Antibody
- 7076 Anti-mouse IgG, HRP-linked Antibody
- 9803 Cell Lysis Buffer (10X)
- 9808 Phosphate Buffered Saline (PBS-20X)
- 9809 Phosphate Buffered Saline with Tween 20 (PBST-20X)
- 9998 BSA
- 7004 TMB Substrate
- 7002 STOP Solution
For Research Use Only. Not For Use In Diagnostic Procedures.