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ReadiLink™ xtra Rapid XFD488 Antibody Labeling Kit *BSA-Compatible, XFD488 Same Structure to Alexa Fluor™ 488*
XFD488 is manufactured by AAT Bioquest, and it has the same chemical structure of Alexa Fluor® 488 (Alexa Fluor® is the trademark of ThermoFisher). ReadiLink™ xtra rapid antibody labeling kits require essentially only 2 simple mixing steps without a column purification needed. Specially formulated and preactivated XFD488 (chemically equivalent to Alexa Fluor® 488) used in this ReadiLink™ kit is quite stable and shows good reactivity and selectivity with antibodies. The kit has all the essential components for labeling ~2x50 ug antibody. Each of the two vials of preactivated XFD488 dye provided in the kit is optimized for labeling ~50 µg antibody. ReadiLink™ xtra XFD488 rapid antibody labeling kit provides a convenient and robust method to label monoclonal and polyclonal antibodies with green fluorescent XFD488 fluorophore. XFD488 is one of the most used fluorophores for labeling antibodies.
Example protocol
AT A GLANCE
Important
Warm all the components and centrifuge the vials briefly before opening, and immediately prepare the required solutions before starting your conjugation. The following protocol is for recommendation.PREPARATION OF WORKING SOLUTION
Protein working solution (Solution A)
For labeling 50 µg of protein (assuming the target protein concentration is 1 mg/mL), mix 5 µL (10% of the total reaction volume) of Reaction Buffer (Component B) with 50 µL of the target protein solution. Note: If you have a different protein concentration, adjust the protein volume accordingly to make ~50 µg of protein available for your labeling reaction. Note: For labeling 100 µg of protein (assuming the target protein concentration is 1 mg/mL), mix 10 µL (10% of the total reaction volume) of Reaction Buffer (Component B) with 100 µL of the target protein solution. Note: The protein should be dissolved in 1X phosphate buffered saline (PBS), pH 7.2 - 7.4; if the protein is dissolved in glycine buffer, it must be dialyzed against 1X PBS, pH 7.2 - 7.4, or use Amicon Ultra-0.5, Ultracel-10 Membrane, 10 kDa (cat# UFC501008 from Millipore) to remove free amines or ammonium salts (such as ammonium sulfate and ammonium acetate) that are widely used for protein precipitation. Note: Impure antibodies or antibodies stabilized with bovine serum albumin (BSA) with 0.1 to 0.5 % will be labeled well. Note: For optimal labeling efficiency, a final protein concentration range of 1 - 2 mg/mL is recommended, with a significantly reduced conjugation efficiency at less than 1 mg/mL.SAMPLE EXPERIMENTAL PROTOCOL
Run conjugation reaction
- Add the protein working solution (Solution A) to ONE vial of labeling dye (Component A), and mix them well by repeatedly pipetting for a few times or vortex the vial for a few seconds. Note: If labeling 100 µg of protein, use both vials (Component A) of labeling dye by dividing the 100 µg of protein into 2 x 50 µg of protein and reacting each 50 µg of protein with one vial of labeling dye. Then combine both vials for the next step.
- Keep the conjugation reaction mixture at room temperature for 30 - 60 minutes. Note: The conjugation reaction mixture can be rotated or shaken for longer time if desired.
Stop Conjugation reaction
- Add 5 µL (for 50 µg protein) or 10 µL (for 100 µg protein) which is 10% of the total reaction volume of TQ™-Dyed Quench Buffer (Component C) into the conjugation reaction mixture; mix well.
- Incubate at room temperature for 10 minutes. The labeled protein (antibody) is now ready to use.
Storage of Protein Conjugate
The protein conjugate should be stored at > 0.5 mg/mL in the presence of a carrier protein (e.g., 0.1% bovine serum albumin). For longer storage, the protein conjugates could be lyophilized or divided into single-used aliquots and stored at ≤ –20°C.Spectrum
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References
View all 50 references: Citation Explorer
TiO2 Nanoparticles and Commensal Bacteria Alter Mucus Layer Thickness and Composition in a Gastrointestinal Tract Model.
Authors: Limage, Rhodesherdeline and Tako, Elad and Kolba, Nikolai and Guo, Zhongyuan and García-Rodríguez, Alba and Marques, Cláudia N H and Mahler, Gretchen J
Journal: Small (Weinheim an der Bergstrasse, Germany) (2020): e2000601
Authors: Limage, Rhodesherdeline and Tako, Elad and Kolba, Nikolai and Guo, Zhongyuan and García-Rodríguez, Alba and Marques, Cláudia N H and Mahler, Gretchen J
Journal: Small (Weinheim an der Bergstrasse, Germany) (2020): e2000601
Long-term treatment with naproxen changes the chemical coding of the porcine intramural duodenum neurons.
Authors: Czajkowska, Marta and Rychlik, Andrzej and Całka, Jarosław
Journal: Annals of anatomy = Anatomischer Anzeiger : official organ of the Anatomische Gesellschaft (2020): 151425
Authors: Czajkowska, Marta and Rychlik, Andrzej and Całka, Jarosław
Journal: Annals of anatomy = Anatomischer Anzeiger : official organ of the Anatomische Gesellschaft (2020): 151425
Conformational Switching in Bcl-xL: Enabling Non-Canonic Inhibition of Apoptosis Involves Multiple Intermediates and Lipid Interactions.
Authors: Vasquez-Montes, Victor and Kyrychenko, Alexander and Vargas-Uribe, Mauricio and Rodnin, Mykola V and Ladokhin, Alexey S
Journal: Cells (2020)
Authors: Vasquez-Montes, Victor and Kyrychenko, Alexander and Vargas-Uribe, Mauricio and Rodnin, Mykola V and Ladokhin, Alexey S
Journal: Cells (2020)
Tracking the physical stability of fluorescent-labeled mAbs under physiologic in vitro conditions in human serum and PBS.
Authors: Schuster, Joachim and Mahler, Hanns-Christian and Koulov, Atanas and Joerg, Susanne and Racher, Andy and Huwyler, Joerg and Detampel, Pascal and Mathaes, Roman
Journal: European journal of pharmaceutics and biopharmaceutics : official journal of Arbeitsgemeinschaft fur Pharmazeutische Verfahrenst (2020): 193-201
Authors: Schuster, Joachim and Mahler, Hanns-Christian and Koulov, Atanas and Joerg, Susanne and Racher, Andy and Huwyler, Joerg and Detampel, Pascal and Mathaes, Roman
Journal: European journal of pharmaceutics and biopharmaceutics : official journal of Arbeitsgemeinschaft fur Pharmazeutische Verfahrenst (2020): 193-201
Generation of biparatopic antibody through two-step targeting of fragment antibodies on antigen using SpyTag and SpyCatcher.
Authors: Akiba, Hiroki and Takayanagi, Kensuke and Kusano-Arai, Osamu and Iwanari, Hiroko and Hamakubo, Takao and Tsumoto, Kouhei
Journal: Biotechnology reports (Amsterdam, Netherlands) (2020): e00418
Authors: Akiba, Hiroki and Takayanagi, Kensuke and Kusano-Arai, Osamu and Iwanari, Hiroko and Hamakubo, Takao and Tsumoto, Kouhei
Journal: Biotechnology reports (Amsterdam, Netherlands) (2020): e00418
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