| Product Includes | Product # | Quantity | Color | Storage Temp |
|---|---|---|---|---|
| Phospho-CrkL (Tyr207) Rabbit mAb Coated Microwells | 17435 | 96 tests |
|
+4C |
| CrkL Rabbit Detection mAb (Biotinylated) | 84578 | 1 ea |
|
+4C |
| HRP-Linked Streptavidin (ELISA Formulated) | 11805 | 1 ea |
|
+4C |
| Detection Antibody Diluent | 13339 | 11 ml |
|
+4C |
| HRP Diluent | 13515 | 11 ml |
|
+4C |
| TMB Substrate | 7004 | 11 ml |
|
+4C |
| STOP Solution | 7002 | 11 ml |
|
+4C |
| Sealing Tape | 54503 | 2 ea |
|
+4C |
| ELISA Wash Buffer (20X) | 9801 | 25 ml |
|
+4C |
| ELISA Sample Diluent | 11083 | 25 ml |
|
+4C |
| Cell Lysis Buffer (10X) | 9803 | 15 ml |
|
-20C |
*The microwell plate is supplied as 12 8-well modules - Each module is designed to break apart for 8 tests.
Description
The PathScan® Phospho-CrkL (Tyr207) Sandwich ELISA Kit is a solid phase sandwich enzyme-linked immunosorbent assay (ELISA) that detects endogenous levels of CrkL protein phosphorylated at Tyr207. A phospho-CrkL (Tyr207) rabbit antibody has been coated onto the microwells. After incubation with cell lysates, phospho-CrkL (Tyr207) protein is captured by the coated antibody. Following extensive washing, a biotinylated CrkL rabbit detection antibody is added to detect the captured phospho-CrkL (Tyr207) protein. HRP-linked streptavidin 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 CrkL protein phosphorylated at Tyr207.
*Antibodies in this kit are custom formulations specific to kit.
Specificity/Sensitivity
Background
CrkL, a 39 kDa adaptor protein, has a key regulatory role in hematopoietic cells. CrkL has one SH2 and two SH3 domains, with 60% homology to CrkII (1). The amino-terminal SH3 domain of CrkL binds proteins, such as C3G, SOS, PI3K, c-Abl, and BCR/Abl. The SH2 domain of CrkL can bind to tyrosine-phosphorylated proteins, such as Cbl, HEF1, CAS, and paxillin (2,3). CrkL is involved in various signaling cascades initiated by different cytokines and growth factors. The biological outcomes of the Crk-activated signal transduction include the modulation of cell adhesion, cell migration, and immune cell responses (4). CrkL is a prominent substrate of the BCR/Abl oncoprotein in chronic myelogenous leukemia and binds to both BCR/Abl and c-Abl (5). CrkL is prominently and constitutively tyrosine phosphorylated in CML neutrophils and is not phosphorylated in normal neutrophils. Moreover, stimulation of normal neutrophils with cytokines and agonists does not induce tyrosine phosphorylation of this protein (6), indicating that it may be a useful target for therapeutic intervention or as a disease marker. Tyr207 in CrkL is the BCR/Abl phosphorylation site (7).
- Satter, M. and Salgia, R. (1998) Leukemia 12, 637-644.
- Feller, S. M. et al. (1998) J. Cell. Physiol. 177, 535-552.
- Kiyokawa, E. et al. (1997) Crit. Rev. Oncog. 8, 329-342.
- Feller, S. M. et al. (2001) Oncogene 20, 6348-6371.
- Grumbach, I. M. et al. (2001) Br. J. Haematol. 112, 327-336.
- Nicholas, G. L. et al. (1994) Blood 84, 2912-2918.
- de Jong, R. et al. (1997) Oncogene 14, 507-513.
Background References
Cross-Reactivity Key
H: human M: mouse R: rat Hm: hamster Mk: monkey Vir: virus Mi: mink C: chicken Dm: D. melanogaster X: Xenopus Z: zebrafish B: bovine Dg: dog Pg: pig Sc: S. cerevisiae Ce: C. elegans Hr: horse GP: Guinea Pig Rab: rabbit All: all species expected
Trademarks and Patents
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