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GFP sdAb - FluoTag-X4 - N0304-AF647-L

GFP is a widely used green fluorescent protein tag

 

This product was developed by   

NanoTag-Biotechnologies

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

Cat. No.: N0304-AF647-L
Amount: 200 µl
Price: $515.00
Cat. No. N0304-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: λex 568 nm / λem 665 nm (identical to Alexa® 568)
Alexa® 647: λex 650 nm / λem 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: not tested yet
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) 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.

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 two FluoTags each
Clone 1H1-1B2
Subtype single domain
Immunogen Recombinant protein corresponding to AA 1 to 238 from jellyfish GFP (UniProt Id: P42212)
Specificity Recognizes GFP (green fluorescent protein) and common GFP derivatives like EGFP, mEGFP, Sirius, tSapphire, Cerulean, eCFP, mTurquoise, acGFP, Emerald, superecliptic pHluorin, paGFP, superfolder GFP, eYFP, mVenus and Citrine

Cat. No.: N0304-AF647-L
Amount: 200 µl
Price: $515.00
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.