PROVETOP

Biotinylated Human Notch 1 Protein

Catalog No: NOTCH1-HB001

Species
Human
Expression System
HEK293
Tag
His, Avi
Activity
Activity verified

Product overview

Recombinant Biotinylated Human Notch 1 Protein is expressed in HEK293 cells with a His tag and Avi tag at the C-terminus. It contains amino acid residues Ala19-Gln526 (UniProt accession: P46531).

Product Details

Molecular Aliases
Notch-1; TAN1; NOTCH1; 9930111A19Rik; lin-12; Mis6; N1;lin12
Protein Length
Ala19-Gln526
Expression System
HEK293
Theoretical Molecular Weight
The protein has a predicted MW of 56.5 kDa. Due to glycosylation, the protein migrates to 70-80 kDa based on Bis-Tris PAGE result.
Purity
> 95% as determined by Bis-Tris PAGE > 90% as determined by HPLC
Endotoxin
Less than 1 EU per μg by the LAL method.
Buffer / Formulation
Lyophilized from 0.22μm filtered solution in PBS (pH 7.4). Normally 8% trehalose is added as protectant before lyophilization.
State
Lyophilized
Storage Conditions
-20 to -80°C for 12 months as supplied from date of receipt. -80°C for 3 months after reconstitution. Recommend to aliquot the protein into smaller quantities for optimal storage. Please minimize freeze-thaw cycles.
Reconstitution Advice
Dissolve the lyophilized protein in distilled water. Please refer to the Certificate of Analysis for detailed instructions.

Data Display

Bis-Tris PAGE
NOTCH1-HB001 Bis-Tris-PAGE-white result

Biotinylated Human Notch 1 on Bis-Tris PAGE under reduced conditions. The purity is greater than 95%.

SEC-HPLC
NOTCH1-HB001 SEC-HPLC result

The purity of Biotinylated Human Notch 1 is greater than 90% as determined by SEC-HPLC.

ELISA
NOTCH1-HB001 ELISA result

Immobilized Biotinylated Human Notch 1, His-Avi tag at 2μg/ml (100μl/well) on the streptavidin precoated plate (5μg/ml). Dose response curve for Human DLL4, hFc Tag with the EC50 of 9.0ng/ml determined by ELISA (QC Test).

SPR
NOTCH1-HB001 SPR result

Human DLL4, hFc Tag captured on CM5 Chip via Protein A can bind Biotinylated Human Notch 1, His Tag with an affinity constant of 0.31 μM as determined in SPR assay (Biacore T200).

SPR
NOTCH1-HB001 SPR-2 result

Biotinylated Human Notch 1, His-Avi tag immobilized on CM5 Chip can bind Human DLL1, His Tag (Cat. DLL-HM101) with an affinity constant of 7.33 μM as determined in SPR assay (Biacore S200).

Background

Human Notch-1 is a 300 kDa type I transmembrane glycoprotein that is one of four human Notch homologues involved in developmental processes. Notch-1 functions as a receptor for membrane-bound ligands Jagged-1 (JAG1), Jagged-2 (JAG2) and Delta-1 (DLL1) to regulate cell-fate determination. Upon ligand activation through the released notch intracellular domain (NICD) it forms a transcriptional activator complex with RBPJ/RBPSUH and activates genes of the enhancer of split locus. Affects the implementation of differentiation, proliferation and apoptotic programs.

References

  1. Hales E C, Taub J W, Matherly L H. New insights into Notch1 regulation of the PI3K–AKT–mTOR1 signaling axis: Targeted therapy of γ-secretase inhibitor resistant T-cell acute lymphoblastic leukemia[J]. Cellular Signalling, 2014, 26(1):149-161.

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