Biotin Antibody Labeling Kit

To perform multiplex analysis for immunofluorescence (IF) or immunohistochemistry, antibody conjugates are often used but are not always commercially available. As an alternative option, and to support their research, scientists often create their own conjugates through selecting carrier free antibodies and conjugation kits which allows for flexibility to tailor design a set of conjugates (correct combination of antibodies with fluors, enzymes or biotin) . The easy to use biotin antibody labeling kits conjugates 50-100 μg of purified antibody (0.5-1.0 mg/mL) to biotin in 10 minutes. The kit also includes a ready-to-use spin filter to rapidly remove >80% of biotin from the conjugated antibody, with minimal loss (optional step) . All components required to label an antibody are included in the kit. Please note - for high labeling efficiency, the starting antibody concentration should be 0.5-1.1mg/ml in buffer free of extraneous proteins such as BSA, serum or gelatin. Antibody formulations containing azide, trehalose or other sugars are acceptable. No tris buffer. Benefits of the kit include: Start to finish time is ~10 minutes and requires 15-30 seconds hands-on time. Antibody yield 95% to 10%. The kit is extremely easy to use - no prior conjugation experience is needed. Select from a diverse range of antibody labels. All components are included in each kit.

Antibody Preparation: 1) Add 1/10 volume of 10X modifier directly to your antibody tube (i.e. for 100 μl of antibody, add 10 μl of modifier) . Labeling with biotin 2) Add antibody solution to the biotin tube, mix thoroughly with pipette or vortex. Incubate in the dark at room temperature for 10 minutes. Final buffer step 3) After the 10-minute incubation is complete, add 90 μL final buffer diluent to biotin/fluor-labeled antibody, vortex, and it is now ready to use. Final antibody concentration is ~0.25-0.5 mg/mL, depending on starting concentration. Notes • For biotin conjugation it is highly recommended for optimal performance to proceed to the PURIFICATION STEP below following the 10-minute incubation with final buffer. Optional Purification Step (a) PBS/azide desalting column preparation - Break off bottom of RED-capped column and loosen the RED cap, place column in a collection tube, then place inmicrocentrifuge and spin for 2 minutes at 1500 g. Discard flow through. (b) Transfer probe labeled mixture to the RED-capped column, place column in a new collection tube and centrifuge for 2 minutes at 1500 g. (c) Transfer desalted fluor labeled antibody to a new capped tube for 4oC storage and you now have a biotin free, labeled sample ready to use.