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Violet 450 Antibody Labeling Kit
低温
2 Labelings
Component A: mFluor™ Violet 450 | 2 vials (One vial is for 50 μg protein) |
Component B: Reaction Buffer | 1 vial (20 µL) |
Component C: TQ™-Dyed Quench Buffer | 1 vial (20 µL) |
Table 1. Available fluorophores in AAT Bioquest ReadyLink™ Rapid Antibody Labelling Kits
Cat# | Labels | Ex (nm) | Em (nm) |
1100 | mFluor™ Violet 450 | 403 | 454 |
1105 | mFluor™ Violet 420 | 398 | 411 |
1110 | mFluor™ Violet 510 | 414 | 508 |
1114 | mFluor™ Violet 540 | 399 | 550 |
1120 | mFluor™ Blue 570 | 553 | 570 |
1123 | mFluor™ Green 620 | 522 | 617 |
1126 | mFluor™ Yellow 630 | 561 | 630 |
1130 | mFluor™ Red 700 | 657 | 700 |
1131 | mFluor™ Red 780 | 629 | 780 |
1220 | iFluor™ 350 | 345 | 442 |
1227 | iFluor™ 555 | 559 | 569 |
1230 | iFluor™ 594 | 592 | 614 |
1235 | iFluor™ 647 | 654 | 674 |
1240 | iFluor™ 680 | 682 | 701 |
1245 | iFluor™ 700 | 693 | 713 |
1250 | iFluor™ 750 | 753 | 779 |
1255 | iFluor™ 488 | 491 | 514 |
1260 | iFluor™ 633 | 638 | 655 |
1265 | iFluor™ 790 | 782 | 811 |
1290 | Cy3 | 555 | 565 |
1292 | Cy5 | 644 | 665 |
1294 | Cy7 | 749 | 776 |
1299 | FITC | 494 | 520 |
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.
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) 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.
Run conjugation reaction
Stop Conjugation reaction
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.
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