Synaptic Systems - GFP sdAb

GFP sdAb - FluoTag-Q - N0301-At488-L

GFP is a widely used green 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

Green fluorescent protein GFP and its derivates have become very popular and universal tools in cell biology. It is a monomeric and fast maturating protein with high photostability. Due to its sensitivity to pH changes it can be used as a biological pH indicator.

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

In FluoTag®-Q each fluorophore is coupled to exactly one FluoTag, which in turn binds to its target molecule in a monovalent fashion. The high binding affinity and a coupling efficiency of > 95% guarantees a highly linear relation between the number of target molecules and the intensity of fluorescence. This enables a direct count of the target molecule of interest. The fluorophore is located exceptionally close to the recognized epitope (< 1.5 nm), which is ideal for all microscopy techniques.

In FluoTag®-X two fluorophore molecules are site-specifically coupled to each FluoTag molecule. Therefore, the reagent simultaneously targets up to four fluorophores (in X4 variants) to the protein of interest, which ensures extra-bright 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 ) Hussain A et al., Elife (2018)
2 ) van Roessel P et al., Nat. Cell Biol. (2002)
3 ) Stephens DJ et al., J. Cell. Sci. (2001)
4 ) Chamberlain C et al., Traffic (2000)

Cat. No. N0301-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 form light!
Applications	WB: not recommended IP: N/A ICC: 1 : 1000 gallery IHC: not tested yet IHC-P: not tested yet 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) 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. 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.
Label	ATTO 488, one fluorophore coupled to one FluoTag
Clone	1H1
Subtype	single domain
Immunogen	Recombinant protein corresponding to AA 1 to 238 from jellyfish GFP (UniProt Id: P42212)
Specificity	Recognizes GFP, mEGFP, superfolder GFP, most common CFP and YFP variants. Does not cross-react with mCherry, mRFP, dsRed, mTagBFP or their most common derivatives.