Synaptic Systems - anti-mScarlet-i sdAb

anti-mScarlet-i sdAb - FluoTag-X2 - N1302-AF647-L

mScarlet is a bright monomeric red fluorescent protein tag

This product was developed by

Camelid single domain antibodies (sdAbs) consist only of one antigen binding site of an Alpaca heavy chain antibody. With only ~15 kDa, these Tags are about 10-times smaller than conventional IgG antibody molecules.

Camelid single domain antibody

Background

mScarlet-i is a basic, very bright red fluorescent protein, derived from a synthetic construct (1). It is reported to be a rapidly-maturing, constitutively fluorescent monomer with moderate acid sensitivity. It is a useful acceptor in Foerster resonance energy transfer (FRET) based imaging techniques (2).

Unlabeled variants and several modifications of sdAbs like biotin, fluorophore or DBCO conjugation are available.

In FluoTag®-X2 two fluorophore molecules are site-specifically coupled to each FluoTag molecule. Therefore, the reagent simultaneously targets two fluorophores to the protein of interest, which ensures up to two-fold („2X“)-brighter signals. Owing to the small size of the FluoTags, the distance between the target epitope and each fluorophore is ~ 3 nm.
In comparison to detection systems using conventional antibodies, FluoTag-X can thus improve the localization accuracy by 10-15 nm. Both features - superior brightness and precise fluorophore placement - render the FluoTag-X products excellent tools for all microscopy techniques.

1 ) Bindels DS et al., Nat. Methods (2017)
2 ) McCullock TW et al., PLoS One (2020)

Cat. No. N1302-AF647-L	200 µl purified antibody, lyophilized from PBS, fluorescence-labeled with Alexa 647. Fluorescence labeled antibodies conjugated to Alexa^® dyes are well suited for standard epi-fluorescence setups and confocal microscopy. AZdye^® 568: λ_ex 568 nm / λ_em 665 nm (identical to Alexa^® 568) Alexa^® 647: λ_ex 650 nm / λ_em 665 nm This product or portions thereof is manufactured under license from Life Technologies Corporation. Alexa Fluor is a registered trademark of Life Technologies Corporation. Purchase of reagents related to the Alexa Fluor technology from Synaptic Systems GmbH provides a license for non-profit and in-house research use only. Any application of above mentioned technology for commercial purpose requires a separate license from Life Technologies Corporation. Albumin was added for stabilization. For reconstitution add 200 µl H₂O. Either add 1:1 (v/v) glycerol, then aliquot and store at -20°C until use, or store aliquots at -80°C without additives. Reconstitute immediately upon receipt! Avoid bright light when working with the antibody to minimize photo bleeching of the fluorescent dye.
Storage	Up to three months: -20°C Up to 12 months: -80°C or below Protect from light!
Applications	WB: not recommended IP: N/A ICC: 1 : 500 IHC-P: not tested yet FACS: yes Western blot (WB); separation of proteins by PAGE and subsequent transfer to a membrane. Detection of target molecules is carried out with antibodies. Some antibodies require special sample preparation steps. For details, please refer to the “Remarks” section. Immunoprecipitation (IP); Immunoisolation or pulldown of a target molecule using an antibody. For details and product specific hints, please refer to the ”Remarks” section. 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-P) of formalin fixed, paraffin embedded (FFPE) tissue (some antibodies require special antigen retrieval steps, please refer to the ”Remarks” section). Immunoreactivity is usually revealed by fluorescence or a chromogenic substrate. Fluorescence-activated cell sorting (FACS); usually cells are fluorescently labeled with antibodies against cell-surface antigens followed by cell-sorter analysis.
Label	Alexa 647, two fluorophores coupled to one FluoTag
Clone	2B12
Subtype	single domain
Immunogen	Recombinant protein corresponding to AA 1 to 232 from sea anemone mScarlet-i
Specificity	Specific for mScarlet-i and most common RFP derivatives