alexa Fluorescent Products | OMICS International
ISSN : 2153-2435
Pharmaceutica Analytica Acta
Like us on:
Make the best use of Scientific Research and information from our 700+ peer reviewed, Open Access Journals that operates with the help of 50,000+ Editorial Board Members and esteemed reviewers and 1000+ Scientific associations in Medical, Clinical, Pharmaceutical, Engineering, Technology and Management Fields.
Meet Inspiring Speakers and Experts at our 3000+ Global Conferenceseries Events with over 600+ Conferences, 1200+ Symposiums and 1200+ Workshops on
Medical, Pharma, Engineering, Science, Technology and Business

Fluorescent Products

Hideharu Shintani*

Chuo University, School of Science, 1-13-27, Kasuga Bunkyo 112-0003 Tokyo, Japan

*Corresponding Author:
Hideharu Shintani
Chuo University, School of Science, 1-13-27
Kasuga Bunkyo 112-0003 Tokyo, Japan
Tel: +81425922336
E-mail: [email protected]

Citation: Shintani H (2013) IFluorescent Products. Pharmaceut Anal Acta 4:e152. doi: 10.4172/2153-2435.1000e152

Copyright: © 2013 Shintani H. This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.

Visit for more related articles at Pharmaceutica Analytica Acta

Introduction

The reaction of lipid peroxidation products, such as malonaldehyde, with amino groups of proteins, free amino acids, aminophospholipids, or nucleic acid bases, produces fluorescent lipid-peroxidation products known as conjugated Shiff bases with the general structure RN=CHCH=CHNHR’ (N,N’-disubstituted 1-amino- 3-iminopropenes) [1]. When these products are excited at 360 nm the fluorescence maximum is in the region 430-440 nm.

Protocol

Spectrofluorimetric analysis of fluorescent lipid peroxidation products is covered in detail elsewhere [2,3]. The method described here is a simple procedure for detection of lipid-soluble and watersoluble fluorescent products formed during lipid peroxidation of biological membranes.

Lipid-Soluble Fluorescent Products

1. Place membrane sample (e.g. oxidized tissue homogenate, plasma, mitochondria, microsomal suspension, or lipoproteins; 0.5 mL) and ethanol-ether (3:1, v/v, 1.5 mL) in screw-capped tubes (1.3×10 cm, Pyrex).

2. Mix on a vortex mixer for 1 min.

3. Centrifuge for 10 min at 3,000 rpm (4°C).

4. Wash the sediment at the bottom of tube twice with ethanolether (3:1, v/v, 2 mL) repeating assay steps 2 and 3.

5. Pipette the solution (1.5 mL) into a 1-mL quartz cuvette.

6. Measure fluorescence (emission) and excitation spectra by spectrofluorimetry. (The fluorescent lipid-peroxidation products have usually an excitation maximum in the region of 355?365 nm and an emission maximum at 430-440 nm).

Water-Soluble Fluorescent Products

1. Dissolve sediments from assay of lipid-soluble fluorescent products (above) in 2.0 mL 15% SDS-PBS (phosphate-buffered saline, pH 7.4) on a vortex mixer for 5 min.

2. Centrifuge for 10 min at 3,000 rpm (room temperature).

3. Pipette the solution (1.5 mL) into a 1-mL quartz cuvette.

4. Measure fluorescence (emission) and excitation spectra by spectrofluorimetry.

Calculation

The ftuorescence intensity of quinine sulfate (1 mg mL-1) in H2SO4 solution (0.05M) is the standard for the relative ftuorescence intensity of the sample [2]. Quinine sulfate in the solution has a ftuorescence maximum at 457 nm when excited at 360nm.

Comments

Fluorescent products which bind to proteins are insoluble in common organic solvents and their structures (ftuorophores) can be disrupted, in part, during extraction of lipids with 2:1 (v/v) chloroformmethanol. They are, however, stable in 3:1 (v/v) ethanol-ether, as described elsewhere [4]. It should be noted that the lipid extracts from biological membranes and tissue homogenates contain retinol as one of interfering ftuorescent compounds. Retinol, a lipid-soluble compound, has a ftuorescence maximum at 478nm when excited at 335 nm [5] and its ftuorescence can be removed from the lipid extracts by exposing it to high-intensity ultraviolet light for 30s [2].

References

Select your language of interest to view the total content in your interested language
Post your comment

Share This Article

Recommended Conferences

  • 7th International Conference on Phamaceutical Auditing
    June 08-09, 2018 Baltimore, USA
  • 4th International Conference on Drug Discovery, Designing and Development
    June 27-28, 2018 Vancouver, Canada
  • 18th World Pharma Congress
    October 18-19, 2018 Warsaw, Poland

Article Usage

  • Total views: 11779
  • [From(publication date):
    August-2013 - Jan 20, 2018]
  • Breakdown by view type
  • HTML page views : 7968
  • PDF downloads : 3811
 

Post your comment

captcha   Reload  Can't read the image? click here to refresh

Peer Reviewed Journals
 
Make the best use of Scientific Research and information from our 700 + peer reviewed, Open Access Journals
International Conferences 2018-19
 
Meet Inspiring Speakers and Experts at our 3000+ Global Annual Meetings

Contact Us

Agri & Aquaculture Journals

Dr. Krish

[email protected]

1-702-714-7001Extn: 9040

Biochemistry Journals

Datta A

[email protected]

1-702-714-7001Extn: 9037

Business & Management Journals

Ronald

[email protected]

1-702-714-7001Extn: 9042

Chemistry Journals

Gabriel Shaw

[email protected]

1-702-714-7001Extn: 9040

Clinical Journals

Datta A

[email protected]

1-702-714-7001Extn: 9037

Engineering Journals

James Franklin

[email protected]

1-702-714-7001Extn: 9042

Food & Nutrition Journals

Katie Wilson

[email protected]

1-702-714-7001Extn: 9042

General Science

Andrea Jason

[email protected]

1-702-714-7001Extn: 9043

Genetics & Molecular Biology Journals

Anna Melissa

[email protected]

1-702-714-7001Extn: 9006

Immunology & Microbiology Journals

David Gorantl

[email protected]

1-702-714-7001Extn: 9014

Materials Science Journals

Rachle Green

[email protected]

1-702-714-7001Extn: 9039

Nursing & Health Care Journals

Stephanie Skinner

[email protected]

1-702-714-7001Extn: 9039

Medical Journals

Nimmi Anna

[email protected]

1-702-714-7001Extn: 9038

Neuroscience & Psychology Journals

Nathan T

[email protected]

1-702-714-7001Extn: 9041

Pharmaceutical Sciences Journals

Ann Jose

[email protected]

1-702-714-7001Extn: 9007

Social & Political Science Journals

Steve Harry

[email protected]

1-702-714-7001Extn: 9042

 
© 2008- 2018 OMICS International - Open Access Publisher. Best viewed in Mozilla Firefox | Google Chrome | Above IE 7.0 version