Tetramethylrhodamine-5-Isothiocyanate (5-TRITC; G isomer) - BioReagent,Biological Stain,for fluorescence analysis,for microscopy,≥90% , CAS No.80724-19-2

CAS: 80724-19-2 Cat. No.: T639043 Molecular Weight: 443.518 PubChem CID: 2762684

GRADE & PURITY BioReagent, Biological Stain, for Fluorescence analysis, for Microscopy, ≥90%

Synonyms

Tetramethylrhodamine-5-Isothiocyanate | 5-TRITC | G isomer

Storage

Protected from light,Store at -20°C

Shipped In

Ice chest + Ice pads

Application

Cell Staining, Fluorescence Analysis

Overview

FITC and TRITC are among the most simple and commonly used reagents for protein fluorescent labeling. These isothiocyanates will crosslink to amino, sulfhydryl, imidazoyl, tyrosyl or carbonyl groups on a protein. However, only the derivatives of primary and secondary amines generally yield stable products. Reactions are most efficient at pH 8-9, and must be performed in an amine-free buffer such as carbonate/bicarbonate. Avoid Tris buffer, which is a primary amine that will compete with the intended labeling reaction. Antibody and other proteins can be effectively labeled with several fluorophore tags per protein molecule when reacted with a 20- to 25-fold molar excess of isothiocyanate-activated fluorophore. Excess nonreacted and hydrolyzed reagent can be removed by dialysis or gel filtration.

Extinction Coeff: 100,000 M⁻¹ cm⁻¹ (at 544 nm in methanol)； Ex/Em Wavelength =541/572 nm

Procedure for Labeling an Antibody with TRITC

This method is adapted from Larsson²

Materials Required

1. TRITC: dissolve in DMSO at 1 mg/ml

2. Conjugation buffer: 100 mM carbonate/bicarbonate buffer, pH 9.0

3. Antibody solution: dialyze antibody into conjugation buffer at 6 mg/ml

Procedure

1. While stirring, slowly add 35 μl of TRITC to 1 ml of the 6 mg/ml antibody solution; mix thoroughly.

2. Incubate for 2 hours at room temperature in the dark.

3. Remove excess and hydrolyzed TRITC by gel filtration.

Additional Information

Fading (photobleaching in tissue sections) can sometimes be reduced by mounting in an alkaline buffered media (pH 9).² There are several reagents that may be used with FITC and/or TRITC derivatives to prevent fading including n-propylgallate at 0.1-0.25 M dissolved in glycerol for FITC or TRITC.³ For FITC derivatives, o- or p-phenylenediamine added to the mounting buffer from 1 μg/ml to 1 mg/ml in glycerin also may be used.²³

References

1. Horisberger, M. 1984. In Immunolabeling for Electron Microscopy. Polak, J., Varndel, I. Ed. Elsevier: Amsterdam, p. 98.

2. Larsson, L. (1988). Immunocytochemistry: Theory and Practice. CRC. Boca Raton, 77-83, 224-225.

3. Goding, J. (1986). Monoclonal Antibodies: Principles and Practice, 2nd ed. Academic, London.

Specifications

Synonyms

Tetramethylrhodamine-5-Isothiocyanate | 5-TRITC | G isomer

Specifications & Purity

BioReagent, Biological Stain, for fluorescence analysis, for microscopy, ≥90%

Stability And Storage

Store at -20℃ long term (36 months). Store in the dark.

Storage

Protected from light, Store at -20°C

Shipped In

Ice chest + Ice pads

This product requires cold chain shipping. Ground and other economy services are not available.

Grade

Biological Stain, BioReagent, for Fluorescence analysis, for Microscopy

Purity

≥90%

Names and Identifiers

• Isomeric SMILES: CN(C)C1=CC2=C(C=C1)C(=C3C=CC(=[N+](C)C)C=C3O2)C4=C(C=C(C=C4)N=C=S)C(=O)[O-]
• PubChem CID: 2762684
• Molecular Weight: 443.518

Chemical and Physical Properties

• Sensitivity: Light-sensitive