Synaptic Systems

Cat. No.: 710 006DY654

Amount: 5 nmol

Price: $355.00

Cat. No. 710 006DY654	5nmol mCLING labeled with DY^® 654 in 100 µl PBS (lyophilized). For reconstitution add 100 µl H₂O, then aliquot and store at -80°C until use. Reconstitute immediately upon receipt! Avoid bright light when working with the probe to minimize photo bleeching of the fluorescent dye.
Applications	ICC: 1 : 50 up to 1 : 250 (1 - 0.2 nmol/ml) gallery IHC: 1 : 25 up to 1 : 50 (2 - 1 nmol/ml) Immunocytochemistry (ICC) on 4% PFA fixed cells. Immunoreactivity is usually revealed by fluorescence. Some antibodies require special fixation methods. For details, please refer to the “Remarks” section. Immunohistochemistry (IHC) on 4% PFA perfusion fixed tissue with 24h PFA post fixation. Immunoreactivity is usually revealed by fluorescence or a chromogenic substrate. Some antibodies require special fixation methods or antigen retrieval steps. For details, please refer to the ”Remarks” section.
Label	DY 654
Remarks	Due to the positive charge of mCLING, negatively charged coatings of cover-slips should be avoided. We recommend a positively charged coating like poly-L-lysine (PLL). mCLING is a fixable dye but paraformaldehyde alone is not able to fix this molecule sufficiently. Therefore, a mixture of 4 %paraformaldehyde (PFA) and 0.2 % glutaraldehyde is strongly advised. For detailed protocols see Revelo NH & Rizzoli SO, 2016.
Data sheet	710_006dy654.pdf

References for mCLING - 710 006DY654

CtBP1-Mediated Membrane Fission Contributes to Effective Recycling of Synaptic Vesicles.
Ivanova D, Imig C, Camacho M, Reinhold A, Guhathakurta D, Montenegro-Venegas C, Cousin MA, Gundelfinger ED, Rosenmund C, Cooper B, Fejtova A, et al.
Cell reports (2020) 307: 2444-2459.e7. 710 006DY654 ICC; tested species: mouse

Cat. No.: 710 006DY654

Amount: 5 nmol

Price: $355.00

ICC

Background

The membrane-binding fluorophore-cysteine-lysine-palmtoyl group (mCLING) is a new probe that selectively binds to the plasma membrane. It is taken up during endocytosis and, in contrast to conventional membrane dyes, remains attached to membranes after fixation and permeabilization and can therefore be combined with immunostaining and super-resolution microscopy.
mCLING was used so far in mammalian-cultured cells, yeast, bacteria, primary cultured neurons, Drosophila melanogaster larval neuromuscular junctions, and mammalian tissue.

1 ) McDonald NA et al., Elife (2017)
2 ) Baranov MV et al., Cell Rep (2016)
3 ) Revelo NH et al., Curr Protoc Neurosci (2016)
4 ) Revelo NH et al., J. Cell Biol. (2014)

Protocols

ICC

IHC

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