Product Pathways - PathScan ELISA
PathScan® Total FLT3 Sandwich ELISA Kit #7202
|7202S||1 Kit (96 assays)||---||In Stock||---|
|7202||carrier free and custom formulation / quantity||email request|
When ordering five or more kits, please contact us for processing time and pricing at firstname.lastname@example.org.
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|Kit Includes||Volume||Solution Color|
|FLT3 mAb coated microwells||96 tests|
|FLT3 Detection Ab||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.
Storage: Kit should be stored at 4°C with the exception of Cell Lysis Buffer, which is stored at –20°C (packaged separately).
CST's PathScan® Total FLT3 Sandwich ELISA Kit is a solid phase sandwich enzyme-linked immunosorbent assay (ELISA) that detects endogenous levels of total FLT3 protein. A FLT3 Mouse mAb has been coated onto the microwells. After incubation with cell lysates, both phospho- and nonphospho-FLT3 proteins are captured by the coated antibody. Following extensive washing, FLT3 Rabbit Antibody is added to detect both the captured phospho- and nonphospho-FLT3 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 total FLT3 protein.
Antibodies in kit are custom formulations specific to kit.
Specificity / Sensitivity
PathScan® Total FLT3 Sandwich ELISA Kit #7202 detects endogenous levels of total FLT3 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 phosphorylated of FLT3 in SEM cells is detected by PathScan® Phospho-FLT3 (Tyr591) Sandwich ELISA kit #7206. In contrast, only a low level of phospho-FLT3 protein is detected in SEM cells treated with CT-53518, an inhibitor of FLT3 tyrosine phosphorylation. The inhibitor does not affect the level of total FLT3 protein detected by PathScan® Total FLT3 Sandwich ELISA kit #7202. Absorbance at 450 nm is shown in the top figure, while the corresponding western blots using Phospho-FLT3 (Tyr591) Antibody #3461 (right panel) or FLT3 Rabbit mAb #3462 (left panel), is shown in the bottom figure.
FMS-related tyrosine kinase 3 (FLT3, also called Flk2), is a member of the type III receptor tyrosine kinase family, which includes c-Kit, PDGFR and M-CSF receptors. FLT3 is expressed on early hematopoietic progenitor cells and supports growth and differentiation within the hematopoietic system (1,2). FLT3 is activated after binding with its ligand FL, which results in a cascade of tyrosine autophosphorylation and tyrosine phosphorylation of downstream targets (3). The p85 subunit of PI3 kinase, SHP2, GRB2 and Shc are associated with FLT3 after FL stimulation (4-6). Tyr589/591 is located in the juxtamembrane region of FLT3 and may play an important role in regulation of FLT3 tyrosine kinase activity. Somatic mutations of FLT3 consisting of internal tandem duplications (ITDs) occur in 20% of patients with acute myeloid leukemia (7).
- Shurin, M.R. et al. (1998) Cytokine Growth Factor Rev 9, 37-48.
- Naoe, T. et al. (2001) Cancer Chemother Pharmacol 48 Suppl 1, S27-30.
- Namikawa, R. et al. (1996) Stem Cells 14, 388-95.
- Beslu, N. et al. (1996) J Biol Chem 271, 20075-81.
- Zhang, S. and Broxmeyer, H.E. (2000) Biochem Biophys Res Commun 277, 195-9.
- Zhang, S. et al. (1999) J Leukoc Biol 65, 372-80.
- Mizuki, M. et al. (2000) Blood 96, 3907-14.
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- 7206 PathScan® Phospho-FLT3 (Tyr591) Sandwich ELISA Kit
- 3474 Phospho-FLT3 (Tyr591) (33G6) Rabbit mAb
- 3461 Phospho-FLT3 (Tyr591) Antibody
- 3466 Phospho-FLT3 (Tyr591) (54H1) Mouse mAb
- 3462 FLT3 (8F2) 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.
PathScan® is a trademark of Cell Signaling Technology, Inc.
Cell Signaling Technology® is a trademark of Cell Signaling Technology, Inc.