Product Pathways - PathScan ELISA
PathScan® Phospho-Axl (panTyr) Sandwich ELISA Kit #7042
|7042C||1 Kit (96 assays)||---||In Stock||---|
|7042||carrier free and custom formulation / quantity||email request|
When ordering five or more kits, please contact us for processing time and pricing at email@example.com.
|Kit Includes||Volume||Solution Color|
|Axl Mouse Ab Coated Microwells||96 tests|
|Phospho Tyrosine Rabbit Detection mAb (7G11)||1 ea||Green (Lyophilized)|
|Anti-rabbit IgG, HRP-linked Antibody (ELISA Formulated)||1 ea||Red (Lyophilized)|
|Detection Antibody Diluent||11 ml||Green|
|HRP Diluent||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.
Storage: Kit should be stored at 4°C with the exception of Lysis Buffer, which is stored at –20°C (packaged separately).
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).
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- 7074 Anti-rabbit IgG, HRP-linked Antibody
- 4939 Axl (C2B12) Rabbit mAb
- 4566 Axl (C44G1) Rabbit mAb
- 8661 Axl (C89E7) Rabbit mAb
- 3269 Axl (L283) Antibody
- 9998 BSA
- 9803 Cell Lysis Buffer (10X)
- 7040 PathScan® Total Axl Sandwich ELISA Kit
- 9808 Phosphate Buffered Saline (PBS-20X)
- 9809 Phosphate Buffered Saline with Tween® 20 (PBST-20X)
- 8954 Phospho-Tyrosine (P-Tyr-1000) MultiMab™ Rabbit mAb mix
- 9411 Phospho-Tyrosine Mouse mAb (P-Tyr-100)
- 9416 Phospho-Tyrosine Mouse mAb (P-Tyr-102)
- 7002 STOP Solution
- 7004 TMB Substrate
This product is intended for research purposes only. The product is not intended to be used for therapeutic or diagnostic purposes in humans or animals.
PathScan is a trademark of Cell Signaling Technology, Inc.
Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc.