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Preactivated Protein Conjugation Reaction

Preactivated Protein Conjugation Reaction

Top panel: sequential sections of mouse retina were stained with anti-NPHP5 antibody (Cat No: 90005), followed by labeling of BiCell Scientific conjugated secondary antibody or Thermo Fisher conjugated secondary antibody. Bottom panel: sequential sections of mouse colon were stained with anti-ZO-1 antibody (Cat No: 00236), followed by labeling of BiCell Scientific conjugated secondary antibody or Thermo Fisher conjugated secondary antibody.

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Preactivated Fluorophore & Enzyme Conjugation Kit

$475.00 – $595.00

Item Cat No.: BCCONJ

Concentration: 1 mg/ml of purified protein

Application: Recombinant Protein Conjugation

Reactivity: Human, Insect, Yeast

BiCell Scientific Inc has developed a series of preactivated protein conjugation kits allowing rapid conjugation of recombinant proteins as well as antibodies to fluorophore or reporter enzyme.

BiCell Scientific’s protein conjugation kit utilized the preactivated succinimidyl ester to conjugate fluorophore or reporter enzyme to target proteins (such as antibodies) that possess exposed amino groups (e.g. N-terminal amino group or ε-amino group). The reaction takes place at pH 9.0 and within concentration range of 0.5-1.0 mg/ml.

Each kit contains 10 pre-packed desalting columns and a full set of buffers to complete the conjugation and purification processes of 5 independent proteins. The kit is able to achieve conjugation on >90% available amino groups in a target protein.

Each kit contains columns, reagents and buffers sufficient for 5 independent conjugation reactions.

  • Preactivated conjugation moiety (0.5 ml)
  • 10 desalting columns (bed volume: 1 ml)
  • Activation buffer (50 ml) (Sodium bicarbonate buffered saline, pH 9.0)
  • Stop buffer (50 ml) (Tris buffered saline, pH 7.5)

Store at 4oC.

For Research Use Only. Not for use in clinical diagnostics.

Step 1. Activation of target protein

  • Centrifuge desalting column at 1,000 xg for 5min;
  • Add 1ml of Activation buffer and centrifuge at 1,000 xg for 5min, and repeat 3 times;
  • Add 1ml of target protein (0.5-1.0 mg/ml) and centrifuge at 1,000 xg for 5min;
  • Collect elutant.

Step 2. Conjugation reaction

  • Add 100μl of preactivated conjugation moiety to the eluted target protein solution;
  • Rotate at room temperature for 30min.

Step 3. Purification of conjugated protein

  • Centrifuge desalting column at 1,000 xg for 5min;
  • Add 1ml of Stop buffer and centrifuge at 1,000 xg for 5min, and repeat 3 times;
  • Add the conjugation reaction solution and centrifuge at 1,000 xg for 5min;
  • Collect elutant.

Additional information

Conjugation labels

Alexa Fluor 488, Alexa Fluor 594, Alexa Fluor 647, Horseradish Peroxidase, Alkaline Phosphatase

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