Product Pathways - PathScan ELISA
PathScan® Phospho-Axl (panTyr) Sandwich ELISA Kit #7042
When ordering five or more kits, please contact us for processing time and pricing at firstname.lastname@example.org.
|Kit Includes||Volume||Solution Color|
|Axl Mouse Ab Coated Microwells||96 tests|
|Phospho-Tyrosine Rabbit Detection Antibody||11 ml||Green|
|Anti-rabbit 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|
|ELISA 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® Phospho-Axl (panTyr) Sandwich ELISA Kit is a solid phase sandwich enzyme-linked immunosorbent assay (ELISA) that detects endogenous levels of tyrosine-phosphorylated Axl protein. An Axl mouse antibody has been coated on the microwells. After incubation with cell lysates, Axl protein (phospho and nonphospho) is captured by the coated antibody. Following extensive washing, a phospho-tyrosine rabbit antibody is added to detect captured tyrosine-phosphorylated Axl protein. Anti-rabbit 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 Axl protein phosphorylated on tyrosine residues.Antibodies in kit are custom formulations specific to kit.
Specificity / Sensitivity
CST's PathScan® Phospho-Axl (panTyr) Sandwich ELISA Kit #7042 detects endogenous levels of tyrosine-phosphorylated Axl 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 Axl in NCI-H2347 cells lysed in the presence of phosphatase inhibitors* (phospho lysate) is detected by PathScan® Phospho-Axl (panTyr) Sandwich ELISA Kit #7042 (top, right). In contrast, a low level of phospho-Axl protein is detected in NCI-H2347 cells lysed in the absence of phosphatase inhibitors* (nonphospho lysate). Similar levels of total Axl protein from both nonphospho or phospho lysates are detected by PathScan® Total Axl Sandwich ELISA Kit #7040 (top, left). Absorbance at 450 nm is shown in the top figure while corresponding western blots using a phospho-Axl (Tyr814) rabbit antibody (right) or a total Axl (C44G1) Rabbit mAb #4566 (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. NCI-H2347 cells were cultured (85% confluence) and lysed with or without the addition of phosphatase inhibitor to the lysis buffer (phospho or nonphospho lysate).
Axl, Sky, and Mer are three members of a receptor tyrosine kinase (RTK) family that share a conserved intracellular tyrosine kinase domain and an extracellular domain similar to those seen in cell adhesion molecules. These RTKs bind the vitamin K-dependent protein growth-arrest-specific 6 (Gas6), which is structurally related to the protein S anticoagulation factor (1). Upon binding to its receptor, Gas6 activates phosphatidylinositol 3-kinase (PI3K) and its downstream targets Akt and S6K, as well as NF-κB (2,3). A large body of evidence supports a role for Gas6/Axl signaling in cell growth and survival in normal and cancer cells (4).
- Crosier, K.E. and Crosier, P.S. (1997) Pathology 29, 131-135.
- Demarchi, F. et al. (2001) J. Biol. Chem. 276, 31738-31744.
- Lee, W. P. et al. (2002) Oncogene 21, 329-336.
- Bellosta, P. et al. (1997) Oncogene 15, 2387-2397.
Have you published research involving the use of our products? If so we'd love to hear about it. Please let us know!
- 7040 PathScan® Total Axl Sandwich ELISA Kit
- 4939 Axl (C2B12) Rabbit mAb
- 4566 Axl (C44G1) Rabbit mAb
- 3269 Axl (L283) Antibody
- 4977 Axl Antibody
- 9411 Phospho-Tyrosine Mouse mAb (P-Tyr-100)
- 9416 Phospho-Tyrosine Mouse mAb (P-Tyr-102)
- 8954 Phospho-Tyrosine (P-Tyr-1000) Rabbit mAb
- 7074 Anti-rabbit 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.