PRINCIPLE

Biotin labeling assay is primarily used for the preparation of biotin-labeled antibody for enzyme immunoassay (EIA). NH2-Reactive Biotin has a succinimidyl ester group, and can easily make a covalent bond with an amino group of the target protein or other macromolecules.

LABELING ANTIBODY WITH BIOTIN

MATERIALS

Reagents

  • NH2-Reactive Biotin
  • Reaction Buffer
  • WS Buffer

Instruments and Consumables

  • Filtration Tube
  • Incubator
  • Microcentrifuge
  • Microtube

METHODS

1. Add 100 μl WS buffer and the sample solution containing 100 μg protein to a Filtration tube.

2. Pipette to mix and centrifuge at 8,000g for 10 min.

3. Add 10 μl DMSO to NH2-Reactive Biotin, and dissolve with pipetting.

4. Add 100 μL reaction buffer to the filtration tube, and then add 8 μL NH2-reactive biotin solution to the filtration tube and pipette to mix.

5. Incubate the tube at 37°C for 10 min.

6. Add 100 μL WS buffer to the filtration tube, and centrifuge at 8,000g for 10 min. Discard the filtrate.

7. Add 200 μL WS buffer to the filtration tube, and centrifuge at 8,000g for 10 min. Repeat this step one more time.

8. Add 200 μL WS buffer, and pipette about 10 times to recover the conjugate. Transfer the solution to a microtube, and store at 0-5°C.

9. Collect the labeled solution. Store at 4°C and protect from light.

HOT PRODUCTS

PDCD1 HIV1 gp120 CD33 IAV HA PRNP TNF MS4A1 CD274 APP EGFR CD40 ERBB3 TFPI ERBB2 SARS-CoV S PCSK9 MET BSG HAVCR2 IL6 CD3 LAG3 ROR1 TNFRSF4 IL33 CD19 CD38 JAG1 IL17A IFNG CD47 HCV E2 Protein IL1A / IL1B HBsAg IGF1R CD4 AXL ANGPT2 IL23A NGF MUC1 TNFRSF18 TNFRSF9 MSTN IL1B HRSV F Protein KDR SORT1 C5 VEGF CXCR4 TNFRSF17 Tau TFRC CTLA4 IL13 IAV H5N1 HA TSLP MSLN SOST
For research use only, not directly for clinical use.
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