Welcome to TGpeptide-Professional peptide manufacturer.

Service hotline:+86-(0)-18115476705

FITC/FAM Labeling

      Fluorescent dyes are the primary means of labeling biomolecules. The most common such dyes are FITC derivatives, which have a wide range of applications in fluorescence microscopy, flow cytometry, and immunofluorescence-based assays. TGpeptide provides many kinds of fluorescent labelings: FITC, FAM, TAMRA, MCA etc. Contact us if your fluorescent dye is not on the list.

  • FITC/FAM Labeling Introduction

         TGpeptide provides fluorescent labelings including FITC, FAM, TAMRA, MCA etc.
          Here are structures for part of them:

          Here we list Ex(nm) and Em(nm) for Fluorescent dyes in following table:

  • Fluorescent dyes

    Ex(nm)

    Em(nm)

    Fluorescent dyes

    Ex(nm)

    Em(nm)

    Hydroxycoumarins

    325

    386

    >R-phycoerythrin
    (PE) (489)

    565

    578

    Dansyl chloride

    340

    578

    Rhodamine Red-X

    560

    580

    AMC

    345

    445

    Tamara

    565

    578

    7-Methoxycoumarin

    360

    410

    Alexa fluor 555

    556

    573

    Alexa fluor

    345

    442

    Alexa fluor 546

    556

    573

    3-Aminocoumarin

    350

    445

    Rox

    575

    602

    Dabcyl

    453

    -

    Alexa fluor 568

    578

    603

    Cy2

    490

    510

    Texas Red

    589

    615

    FAM

    495

    517

    Alexa fluor 594

    590

    617

    Alexa fluor 488

    494

    517

    Alexa fluor

    621

    639

    FITC

    495

    519

    Alexa fluor 633

    650

    668

    Alexa fluor 430

    430

    545

    Cy5 (625)

    650

    670

    5-FAM

    492

    518

    Alexa fluor 660

    663

    690

    Alexa fluor 532

    530

    555

    Cy5.5

    675

    694

    HEX

    535

    556

    TruRed 490

    675

    695

    5-TAMRA

    542

    568

    Alexa fluor 680

    679

    702

    Cy3

    550

    570

    Cy7

    743

    767

    TRITC

    547

    572

    Cy3.5

    581

    596

  • · FITC


          FITC is more reactive than carboxy fluorescein, which must also be activated before use. After the selective unmasking of a protecting group, FITC might react with either lysine, an ornithine side chain, or with a primary amino group at the N-terminus of the growing peptide. In this latter case, an alkyl spacer such as aminohexanoic acid (Ahx) is introduced between the last amino acid and the thiourea linkage that is generated during the reaction between isothiocyanate and amine. FITC can react extensively with sulfhydryl groups, such as the side chains of reduced cysteine residues too.

  • · AMC


          AMC(7-Amino-4-methylcoumarin) is another widely used fluorescein, but AMC is different because it is labeled at C-terminal of peptide:
          (1)AMC reacts with alfa- carboxy of first amino acids at C-terminal(AA-AMC);
          (2)Synthesize the left peptides sequence (from the second amino acid from C-terminal of the peptide) using SPPS;
          (3)Couple AA-AMC what we get in step (1) to full-protected peptides what we get in step (2) using liquid phase reaction;
          (4)Make cleavage for peptide what we get in step(3), then we get AMC labeled peptide.

          Basically, Fluorescent dyes such as biotin and FITC can generally be added to either the N- or C-terminus of a peptide. But we recommend adding such modifications to the N terminus of peptides because it results in shorter turnaround time, is more likely to be successful, and does not require additional coupling steps. The addition of an N-terminal modification is the final step in the SPPS protocol because peptides are synthesized from the C- to the N-terminus. We also recommended that clients include a flexible spacer, such as the six-carbon linker Ahx, in their synthetic peptide projects because it increased the stability of the fluorescent label. FITC can also be linked readily to the thiol of cysteine or an amino group of lysine.

Copyright © Nanjing TGpeptide Biotechnology Co.,Ltd.