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RFP sdAb - N0405-DBCO

A red 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.: N0405-DBCO
Amount: 250 µg
Price: $770.00
Cat. No. N0405-DBCO 250 µg purified antibody, lyophilized from PBS, fluorescence-labeled with DBCO.

Dibenzocyclooctyne (DBCO, ADIBO) is one of the most reactive cycloalkynes for strain promoted alkyne azide cycloaddition (spAAC), a copper-free click chemistry reaction.

Albumin was added for stabilization. For reconstitution add 250 µl H2O to get a 1mg/ml solution in PBS. 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.
Applications
 
ICC: yes
FACS: yes

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.

Fluorescence-activated cell sorting (FACS); usually cells are fluorescently labeled with antibodies against cell-surface antigens followed by cell-sorter analysis.

Label DBCO
Clone 2B12
Immunogen Recombinant protein corresponding to AA 1 to 225 from sea anemone RFP (UniProt Id: Q9U6Y8)
Specificity mRFP and derivatives like mCherry, mScarlet-i, tdTomato, dsRed, mOrange
Cat. No.: N0405-DBCO
Amount: 250 µg
Price: $770.00
Background

Red fluorescent protein RFP is a common fusion protein used in cell biology to follow the localization and expression of proteins of interest. RFP is known for its rapid maturation and high resistance to photobleaching and therefore well suited for multi color imaging.

 

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.