Product Pathways - PathScan ELISA
PathScan® Total Notch1 Sandwich ELISA Kit #7245
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|Kit Includes||Volume||Solution Color|
|Notch1 Rat Antibody Coated Microwells||96 tests|
|Notch1 Rabbit 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.
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.
PathScan® Total Notch1 Sandwich ELISA Kit from Cell Signaling Technology is a solid phase sandwich enzyme-linked immunosorbent assay (ELISA) that detects endogenous levels of Notch1. A Notch1 rat antibody has been coated onto the microwells. After incubation with cell lysates, Notch1 protein is captured by the coated antibody. Following extensive washing, a Notch1 rabbit antibody is added to detect the captured Notch1 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 Notch1.Antibodies in kit are custom formulations specific to kit.
Specificity / Sensitivity
PathScan® Total Notch1 Sandwich ELISA Kit #7245 detects endogenous levels of Notch1 protein 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.
Figure 2. The relationship between the protein concentration of lysates from Jurkat cells, untreated or treated with γ secretase inhibitor, and the absorbance at 450 nm using the PathScan® Total Notch1 Sandwich ELISA Kit #7245 is shown.
ELISA - Western correlation
Figure 1. Both cleaved and noncleaved Notch1 can be detected by the PathScan® Total Notch1 Sandwich ELISA Kit #7245. Jurkat cells express cleaved Notch1 (Val1744) and γ secretase inhibitor inhibits the cleavage of Notch1. Jurkat cells were treated with 25 μM γ secretase inhibitor for 96 hours. The absorbance readings at 450 nm are shown on the top, while the corresponding western blots using Notch1 (D1E11) XP® Rabbit mAb #3608 (left panel) or Cleaved Notch1 (Val1744) Antibody #2421 (right panel) are shown on the bottom.
Notch proteins (Notch1-4) are a family of transmembrane receptors that play important roles in development and the determination of cell fate (1). Mature Notch receptors are processed and assembled as heterodimeric proteins, with each dimer comprised of a large extracellular ligand-binding domain, a single-pass transmembrane domain, and a smaller cytoplasmic subunit (Notch intracellular domain, NICD) (2). Binding of Notch receptors to ligands of the Delta-Serrate-Lag2 (DSL) family triggers heterodimer dissociation, exposing the receptors to proteolytic cleavages; these result in release of the NICD, which translocates to the nucleus and activates transcription of downstream target genes (3-4).
- Artavanis-Tsakonas, S. et al. (1999) Science 284, 770-6.
- Chan, Y.M. and Jan, Y.N. (1998) Cell 94, 423-6.
- Schroeter, E.H. et al. (1998) Nature 393, 382-6.
- Rand, M.D. et al. (2000) Mol Cell Biol 20, 1825-35.
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- 7194 PathScan® Cleaved Notch1 (Val1744) Sandwich ELISA Kit
- 3608 Notch1 (D1E11) XP® Rabbit mAb
- 4380 Notch1 (D6F11) XP® Rabbit mAb
- 4147 Cleaved Notch1 (Val1744) (D3B8) Rabbit mAb
- 7074 Anti-rabbit IgG, HRP-linked Antibody
- 9803 Cell Lysis Buffer (10X)
- 7004 TMB Substrate
- 7002 STOP Solution
For Research Use Only. Not For Use In Diagnostic Procedures.