Synaptic Systems - anti-mScarlet-i sdAb

anti-mScarlet-i sdAb - FluoTag-X2 - N1302-At488-L

mScarlet is a bright monomeric red fluorescent protein tag

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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-At488-L	200 µl purified antibody, lyophilized from PBS, fluorescence-labeled with ATTO^® 488. For many of the fluorescence labeled antibodies conjugated to ATTO^® dyes from ATTO-TEC the established fluorescence detection systems can be used. ATTO^® 488: λ_ex 500 nm / λ_em 520 nm ATTO^® 550: λ_ex 554 nm / λ_em 576 nm ATTO^® 565: λ_ex 564 nm / λ_em 590 nm ATTO^® 594: λ_ex 603 nm / λ_em 626 nm ATTO^® 647N: λ_ex 646 nm / λ_em 664 nm ATTO^®488, ATTO^®565, ATTO^®647N and ATTO^®594 dyes are suitable for Stimulated Emission Depletion (STED) microscopy which allows higher resolution imaging compared to confocal laser scanning microscopy. ATTO^®488, ATTO^®565 and ATTO^®594 are also recommended for PALM and dSTORM high resolution microscopy. This product or portions thereof is manufactured under license from ATTO-TEC GmbH. ATTO is a trademarks of ATTO-TEC GmbH, Siegen/Germany. Purchase of reagents related to the AbberiorStar 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 ATTO-TEC GmbH. 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	ATTO 488, 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