Instructions for use (taking 100μl of pure magnetic beads as an example):
1. Preparation of Streptavidin MagBeads:
(1) Shake or vortex the vial to completely resuspend the magnetic beads.
(2) Transfer 100μl of magnetic beads to a new tube (it is recommended to cut the tip of the pipette at an angle to enlarge the entrance).
(3) Place the test tube on a magnetic rack to collect the magnetic beads and discard the supernatant.
(4) Add 0.5 ml of wash buffer to the test tube and invert the test tube several times to mix. Use a magnetic rack to collect the magnetic beads and discard the supernatant. Repeat this step twice.
Note: When applied to nucleic acids, TES Buffer is recommended for washing buffer; when applied to antibodies/proteins, PBS (pH 7.4) is recommended for washing buffer.
2. Fixation of biotinylated molecules
(1) Related reagents
① Usage ratio: 2-3 mg of biotinylated sample: 1 ml of pure magnetic beads.
② Binding/washing buffer: nucleic acid: TES buffer; protein/antibody: PBS (pH 7.4).
③ Elution buffer: 8 M guanidine•HCl (pH 1.5).
(2) Operation steps
① Resuspend the magnetic beads in 100μl binding/washing buffer.
② Add the biotinylated sample to the magnetic beads prepared in step 1 and gently invert to mix.
③ Incubate the test tube at room temperature for one hour with shaking (oscillator or vortexer).
④ Transfer the test tube to a magnetic rack, collect the magnetic beads, and discard the supernatant. (If necessary, retain the supernatant for analysis)
⑤ Add 1 ml binding/washing buffer to the test tube and mix thoroughly, collect the magnetic beads using a magnetic rack and discard the supernatant, and repeat the wash 3 times.
⑥ Resuspend in an appropriate buffer to the desired concentration for downstream use.
3. Antigen purification
(1) Relevant reagents
① Ratio: 2-3 mg biotinylated antibody: 1 ml pure magnetic beads
② Binding/washing buffer: 0.1 M phosphate buffer (containing 0.15 M NaCl, pH 7.2)
③ Elution buffer: 0.1 M glycine•HCl (pH 2.5-2.8)
④ Neutralization buffer: 1 M Tris•HCl (pH 8.5)
(2) Operation steps
① Add 100 μl binding/washing buffer to the magnetic beads prepared in step 1 and resuspend.
② Add the biotinylated antibody solution to the magnetic beads and gently invert to mix.
③ Incubate the test tube at room temperature for one hour (on an oscillator or vortexer).
④ Transfer the test tube to the magnetic rack, collect the magnetic beads, and discard the supernatant. (If necessary, keep the supernatant for analysis)
⑤ Add 1 ml of binding/washing buffer to the test tube and mix thoroughly, collect the magnetic beads using a magnetic stand and discard the supernatant, and repeat the wash 3 times.
⑥ Add 100μl of binding/washing buffer to resuspend the magnetic beads.
⑦ Add the antigen sample to the test tube, gently invert to mix, and incubate at room temperature for 30 minutes or at 4°C overnight.
⑧ Add 1 ml of binding/washing buffer to the test tube to wash the magnetic beads, collect the magnetic beads using a magnetic stand and discard the supernatant, and repeat the wash 3 times.
⑨ Add 100μl of elution buffer to the test tube, mix thoroughly, and incubate at room temperature for 5 minutes.
⑩ Collect the magnetic beads using a magnetic stand and save the supernatant containing the target antigen.
⑪ Add neutralization buffer to adjust the pH, add 5 μl of neutralization buffer to every 50μl of elution.
Note:
1. The recommended storage temperature of Streptavidin MagBead is 2–8°C. Do not freeze this product!
2. During storage and use, Streptavidin MagBead must be kept in liquid suspension. Drying will seriously interfere with the performance and binding ability of the magnetic beads.
3. Please resuspend the magnetic beads before use.
4. Be careful during the use process to avoid bacterial/fungal contamination.