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Streptavidin（SAV）Particles
Datasheet
Catalog Number:
E16MP003-05Amount:
5.0mlForm of Antibody:
Phosphate-buffered solution, pH 7.4, containing 0.09% sodium azideStorage/Stability:
Store at 4°C.DO NOT FREEZE. LIGHT SENSITIVE MATERIAL.Background:
Streptavidin Particles in combination with primary biotinylated antibodies are ideal for depletion or positive isolation of cells from different species (e.g. mouse, human), depending on the specificity of the primary antibody. Cells can be directly isolated from any sample such as whole blood, bone marrow, MNC suspensions or tissue digests.The primary biotinylated antibodies are either added to the cell sample (indirect technique) or pre-coated onto the beads(direct technique) prior to cell isolation. Streptavidin Particles are then mixed with the cell sample in a tube. Streptavidin Particles bind to the target cells during a short incubation, and then the bead-bound cells are separated by a magnet.Depletion – Discard the bead-bound cells and use the remaining, untouched cells for any application. Positive isolation – Discard the supernatant and use the bead-bound cells for downstream molecular applications.Reactivity:
Human,Not yet tested in other species.Experimental Methods:
1.Sample preparation: when working with anticoagulated peripheral blood should be isolated by density gradient centrifugation, for example, using Ficoll -paque. Add 2ml Isolation Buffer wash three times. Prepare PBMC to 1 × 107cells /ml in Isolation Buffer.
2.Add 0.1-20µg primary biotinylated antibody in 1107 PBMC , Incubate for 30 min at room temperature.
3.Sample tube is set to 1000 rpm centrifugation for 5 minutes, discard the supernatant; add 1 ml Isolation Buffer, discard the supernatant, Repeat 2 times(21ml).
4.Transfer 1 ml cells (1 × 107) to a tube and add 50μL re-suspended Streptavidin Particles. Incubate for 30 min at room temperature. With gentle tilting and rotation.
5.Place the tube in a magnet for 3-5min.
For depletion; transfer supernatant to a new tube for further use and discard the beads.
For positive isolation; while the tube is still in the magnet, carefully remove and discard the supernatant.
6.Remove the tube from the magnet and add 1 ml Isolation Buffer, pipet 2–3 times and place the tube in a magnet for 3-5 min. While the tube is still in the magnet, carefully remove and discard the supernatant.
7.Repeat step 4 at 4-5 times. Wash the bead-bound cells. This step is critical to obtain a high purity of isolated cells.
8.Resuspend the cell pellet in preferred cell medium.
9.Keep the cells on 2°C to 8°C until further use in downstream applications.Notices:
1．Use a mixer that provides tilting and rotation of the tubes to ensure that beads do not settle in the tube.
2．Isolation Buffer: Ca2+ and Mg2+ free PBS supplemented with 0.5% BSA and 2 mM EDTA, 0.09%NaN3, pH 7.4.
3．For research use only, not for diagnostic or therapeutic use.
Price:
530$
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Catalog Number
- Amount:5.0ml
- Price:530$
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