ATTO 647 

ATTO 647 is a carborhodamine fluorophore and imaging tracer with photostable properties. ATTO 647 serves as a fluorescent probe to investigate cell membrane structure and diffusion characteristics. When conjugated with wheat germ agglutinin, ATTO 647 specifically binds to N-acetyl-β-D-glucosamine and sialic acid residues on membrane glycoproteins, enabling single-molecule tracing of glycoprotein diffusion. ATTO 647 exhibits highly stable fluorescence properties with significantly reduced blinking in mounting media such as ROXS (AA/MV) and ROXS (TX/TQ), whereas its brightness properties vary in Ibidi-MM and Vectashield. ATTO 647 can also be used to label histone H2B-GFP in fixed cells for confocal microscopy photobleaching experiments^[1][2].

ATTO 647 (0.3 mg; 2 h) can covalently label the purified WGA, with an average of 1.5 ATTO 647 fluorescent groups binding to each WGA molecule^[1].
ATTO 647 (100 nM) in the Atto 647-WGA conjugate will exhibit single-step or double-step photobleaching when bound to the hyaluronic acid-coated glass cover slip, which is consistent with the situation where each conjugate contains one or two fluorescent groups^[1].

Guide (The following is the experimental plan we recommend. This plan serves only as a reference guide. The specific operations should be adjusted according to your actual needs.)
Application of ATTO 647 in the Stably Expressing Group Protein H2B-GFP Human HeLa Cells for Staining^[2][3]:
1. Cultivate the cells until 50-80% confluence, fix with 3.7% formaldehyde.
2. Triton X-100 0.5% permeabilization, BSA/PBST 2% blocking.
3. Incubate with ATTO 647N conjugated antibody to label the intranuclear group protein H2B-GFP.
4. Seal the sections with the following media: PBS, Vectashield (VS), Ibidi-MM, ROXS (AA/MV) (1 mM ascorbic acid + 1 mM methylene blue + enzymatic deoxygenation system), ROXS (TX/TQ) (1 mM Trolox + UV pre-generated Trolox - quinone + enzymatic deoxygenation system).
5. Observe the results under a 635 nm laser.

MedChemExpress (MCE) has not independently confirmed the accuracy of these methods. They are for reference only.

693.25

C₃₄H₄₅ClN₂O₉S

906664-68-4

O=C(CCCN1C2=C(C(CC1(C)C)C)C=C(C=C3C=C4C(C=C3C5(C)C)=[N+](C(C)(C)C=C4CS(=O)(O)=O)CC)C5=C2)O.O=Cl(=O)([O-])=O

Room temperature in continental US; may vary elsewhere.

Please store the product under the recommended conditions in the Certificate of Analysis.

• [1]. Bruckbauer A, et al. Nanopipette delivery of individual molecules to cellular compartments for single-molecule fluorescence tracking. Biophys J. 2007;93(9):3120-3131.

  [2]. Cordes T, et al. Mechanisms and advancement of antifading agents for fluorescence microscopy and single-molecule spectroscopy. Phys Chem Chem Phys. 2011;13(14):6699-6709.

  [3]. Kocabey S, et al. Cellular Uptake of Tile-Assembled DNA Nanotubes. Nanomaterials (Basel). 2014;5(1):47-60. Published 2014 Dec 30.