ReadiLink™ Rapid XFD488 Antibody Labeling Kit *XFD488 Same Structure to Alexa Fluor™ 488*
|Shipping||Standard overnight for United States, inquire for international|
|Extinction coefficient (cm -1 M -1)||73000|
|H-phrase||H303, H313, H333|
|Intended use||Research Use Only (RUO)|
|R-phrase||R20, R21, R22|
|ReadiLink™ Rapid iFluor® 488 Antibody Labeling Kit *Microscale Optimized for Labeling 50 μg Antibody Per Reaction*|
Extinction coefficient (cm -1 M -1)
AT A GLANCE
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
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) or gelatin will not 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
- 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.
- 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.
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.
|Extinction coefficient (cm -1 M -1)||73000|
Authors: Martín-Fontecha, Mar and Angelina, Alba and Rückert, Beate and Rueda-Zubiaurre, Ainoa and Martín-Cruz, Leticia and van de Veen, Willem and Akdis, Mübeccel and Ortega-Gutiérrez, Silvia and López-Rodríguez, María Luz and Akdis, Cezmi A and Palomares, Oscar
Journal: Bioconjugate chemistry (2018): 382-389
Authors: Amoury, Manal and Blume, Tobias and Brehm, Hannes and Niesen, Judith and Tenhaef, Niklas and Barth, Stefan and Gattenlöhner, Stefan and Helfrich, Wijnand and Fitting, Jenny and Nachreiner, Thomas and Pardo, Alessa
Journal: Current pharmaceutical design (2013): 5429-36
Authors: Girgenrath, Tanya and Mahalingam, Mohana and Svensson, Bengt and Nitu, Florentin R and Cornea, Razvan L and Fessenden, James D
Journal: The Journal of biological chemistry (2013): 16073-84
Authors: Hirose, Hisaaki and Takeuchi, Toshihide and Osakada, Hiroko and Pujals, Sílvia and Katayama, Sayaka and Nakase, Ikuhiko and Kobayashi, Shouhei and Haraguchi, Tokuko and Futaki, Shiroh
Journal: Molecular therapy : the journal of the American Society of Gene Therapy (2012): 984-93
Authors: Lemke, Edward A
Journal: Methods in molecular biology (Clifton, N.J.) (2011): 3-15
Authors: Goto, Hiroaki and Okuda, Satohiro and Mizukami, Akane and Mori, Hitoshi and Sasaki, Narie and Kurihara, Daisuke and Higashiyama, Tetsuya
Journal: Plant & cell physiology (2011): 49-58
Authors: DeKroon, Robert M and Osorio, Cristina and Robinette, Jennifer B and Mocanu, Mihaela and Winnik, Witold M and Alzate, Oscar
Journal: Journal of proteome research (2011): 1632-44
Authors: Anderson, Valerie L and Webb, Watt W
Journal: BMC biotechnology (2011): 125
Authors: Li, Xiuru and Guo, Jun and Asong, Jinkeng and Wolfert, Margreet A and Boons, Geert-Jan
Journal: Journal of the American Chemical Society (2011): 11147-53
Authors: Tomita, Kazunari and Berger, Evelyn J and Berger, Richard A and Kraisarin, Jirachart and An, Kai-Nan
Journal: The Journal of hand surgery (2007): 466-73