Product Datasheet

 
Product Description
 
Product: Human PAI-1 NBD labeled at the reactive center loop
Catalog Number: HPAI-P9NBD
Product Page: https://www.mol-innov.com/products/IHUPAI1RP9NBD500UG
Description: P9-NBD PAI-1 was created by mutagenesis of the P9 serine residue (Ser338) on the reactive center loop to cysteine. This provides a free thiol group for incorporation of N,N'-dimethyl-N-(acetyl)-N'-(7-nitrobenz-2-oxa-1,3-diazol-4-yl) (NBD), a fluorescent probe highly sensitive to changes in solvation and hydrophobic environment. The fluorescence emission of P9-NBD PAI-1 is enhanced 6-7 fold upon insertion of the reactive center loop into beta-sheet A following complex formation with proteinases, formation of the latent species, or cleavage by elastase. The incorporated probe is excited at 480 nm and displays a broad emission spectrum with a peak centered 542 nm with a resultant blue-shift to 520 nm following reactive center loop insertion. The modified PAI-1 is nearly as active as wt PAI-1 and is more resistant to the spontaneous latency reaction making this an excellent tool for monitoring reaction rates of PAI-1 (1). P9-NBD PAI-1 has been utilized in a number of studies to determine the rates of loop insertion and SERPIN reaction mechanisms when reacted with various proteinases (1,2), inactivating antibodies (2) and conformational changes imposed by the binding of vitronectin (4).

References:
1. Shore JD, et al.(1995) J Biol Chem 270:5395-5398.
2. Lawrence D, et al.(2000) J Biol Chem 275:5839-5844.
3. Verhamme IM, et al.(1999) J Biol Chem 274:17511-17517.
4. Gibson A, et al.(1997) J Biol Chem 272:5112-5121.

Recommended Storage: SAMPLE
Shipping Conditions: >95percent active by uPA titration; 1 mole NBD / mole PAI-1
Minimum Shelf Life: 500.50
Source: 2.5 mg/ml
Molecular Weight: Proteins
Extinction Coefficient: 43,000
Antigen: -70 C; Protect from light
Target: Frozen liquid
Target Molecular Weight: PAI-1
Host: 934.5
Clonality: 30
Isotype: E. coli
Clone: s
Applications: Dry ice
Cross-reactivity: 3 years from delivery
 
Sample Characteristics
 
Lot Number: HPAI-P9NBD-IHUPAI1RP9NBD1MG
Aliquot: 1 x Human PAI-1 Recombinant P9-NBD Labeled 1 mg
Concentration: 715
Volume: 1335
Titer: 0.05M Sodium Phosphate; 0.1M NaCl; 1mM EDTA; pH 6.6
Buffer: SAMPLE
Form: 0.5 mg
Preservative: 0.2 ml
 
Optimal experimental conditions and working dilutions must be determined by the end user. Sample concentration and volume are for example purposes only and do not necessarily represent the product that will be received. Please contact us for a current lot-specific product datasheet.
 
For in vitro laboratory use only
 
Molecular Innovations, Inc. - 46430 Peary Court, Novi, MI 48377, USA
Tel: (888) 557-5055 - Fax: (248) 896-0148 - info@mol-innov.com