ReadiUse™ Preactivated PE-iFluor™ 750 Tandem

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<p>Our preactivated PE was premodified with our Buccutite™ FOL (provided). Your antibody (or other proteins) is modified with our Buccutite™ MTA (provided as free sample) to give MTA-modified protein (such as antibody). The MTA-modified protein readily reacts with FOL-modified PE (provided) to give the desired PE-antibody conjugate in much higher yield than the SMCC chemistry. In addition our preactivated PE reacts with MTA-modified biopolymers at much lower concentrations than the SMCC chemistry.</p>
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Unit Size: Cat No: Price (USD): Qty:
1 mg 2578 $345

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Additional Ordering Information
Telephone: 1-800-990-8053
Fax: 1-408-733-1304
International: See distributors


Ex/Em (nm)566/778
Storage Refrigerated (2-8 °C)
Minimize light exposure
Category PE and APC,
Assay and Sample Preparation,
Biomolecule Labeling

R-Phycoerythrin (PE) is isolated from red algae. Its primary absorption peak is at 565 nm with secondary peaks at 496 and 545 nm. Our PE-iFluor™ 750 Tandem is an excellent replacement for PE-Cy7® and PE-Alexa Fluor® 750 Tandem due to their similar spectral properties. ReadiUse™ Preactivated PE-iFluor™ 750 Tandem is an activated PE protein, and can be easily conjugated to antibodies with much higher conjugation yield than the conventional PE.

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This protocol only provides a guideline, and should be modified according to your specific needs.
  1. Pre-activate Antibody with Buccutite™ MTA
    1. Reconstitute Buccutite™ MTA in DMSO at ~10 mg/mL.
      Note: Please store unused MTA at -20°C and could be used up to two freeze and thaw cycles.
    2. Prepare target antibody (Ab) in pH = 8.5~9.0 buffer at concentration above 1 mg/ml
    3. Add the MTA to Ab solution at the ratio of 8~10 µg MTA/100 µg Ab.
    4. Mix well and react at RT for 60 minutes, rotating during the reaction.
    5. Purify the reaction mixture with desalting column to remove unreacted MTA and exchange buffer to PBS or buffer of your choice.
    6. Collect the MTA-activated Ab, and estimate the concentration by 70% yield of the original starting amount.

  2. Conjugate with Pre-activated PE, PE-iFluors, or PE-iFluor Tandems
    1. Reconstitute pre-activated PE, PE, PE-iFluors, or PE-iFluor Tandems in 100 µL ddH2O to 10mg/mL.
      Note: Reconstituted pre-activated PE, PE-iFluors, or PE-iFluor Tandems are not stable and could be stored at 4 °C for one month, please kept it from light.
    2. Add pre-activated PE, PE-iFluors, or PE-iFluor Tandems directly to MTA-activated target Ab solution (from 1.5) at the ratio of 300 µg PE-iFluor647/100µg MTA-activated Ab.
    3. Rotate the mixture for 1~2 hours at room temperature
    4. The Ab/ PE, PE-iFluors, or PE-iFluor Tandems conjugates are now ready to use.
      Optional: Ab/ PE, PE-iFluors, or PE-iFluor Tandems conjugate could be further purified through size exclusion chromatography to get best performance.
      Note1: The antibody conjugate should be stored at > 0.5 mg/mL in the presence of a carrier protein (e.g., 0.1% bovine serum albumin) and 0.02-0.05% sodium azide.
      Note2: The Ab/ PE, PE-iFluors, or PE-iFluor Tandems conjugate solution could be stored at 4 °C for two months and kept from light.

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