μMACS™ Streptavidin Kit ( #130-074-101 #130-091-287 )

μMACS™ Streptavidin Kit

Versatile—the µMACS™ Streptavidin Kit allows the magnetic isolation of any interacting molecule with a biotinylated probe.

Easily done—due to MACS® Column Technology, the target molecule is magnetically bound to a column for simple washing steps, in-column assays, and high-purity elution.

Gentle—to preserve the interaction of partners, centrifugation, resuspension, or multiple pipetting steps of the sample are avoided.

Specific—a biotinylated, individual interacting molecule, such as a complementary nucleic acid, an antibody, or a functional partner will bind the target.

Any interacting molecule. The µMACS Streptavidin Kit allows the highly specific isolation of any molecule if a biotinylated interaction partner is available.
Such isolations can be used for detection and downstream analysis of proteins, such as receptor ligands or signaling activators as well as nucleic acids, for example, RNA transcripts or viral sequences. Furthermore, large molecular complexes, organelles, or even viable viruses can be purified with MACS® Column Technology. Special protocols for diverse applications are available.

The convenient procedure. A biotinylated probe, such as DNA, RNA, protein, or an antibody specifically binds to the target molecule in the cell lysate or any other molecular suspension. µMACS Streptavidin MicroBeads bind to the biotin-residues, thereby, magnetically labeling the whole complex. Due to the extremely small size of the MACS® MicroBeads (diameter 50 nm), these MicroBeads are colloidal and remain in suspension, while they quickly find their target. When loading the labeled cell lysate on a µ Column placed in the strong magnet of the µMACS or thermoMACS™ Separator the magnetically labeled complex is retained in the column. Subsequent stringent washing steps, simply performed by adding buffer, remove non-specifically bound molecules. Finally, the non-biotinylated target molecules can be eluted with high purity, whereas the biotinylated probe with MicroBeads remains in the column.

Unlimited applications

Gene transcription and translation regulation
The regulation of gene transcription and translation plays a major role in differentiation, disease development, signal transduction, or cell signaling. Transcriptional control is accomplished by the binding of proteins (transcription factors) to specific regulatory DNA sequences. The control of translation and mRNA degradation is similarly regulated by specific proteins binding to RNA sequences. Using biotinylated target DNA or RNA sequences, interacting proteins can be pulled out of a cell lysate (fig.1).^1-6 Further means of regulation include the cell type-specific availability of tRNA molecules that influences the translation rate. With the help of biotinylated complementary sequences, tRNAs can be determined.

Phage display and protein interactions
To elucidate gene function and to decipher signaling pathways, the identification and isolation of protein interactions is crucial. By biotinylating a protein and adding μMACS Streptavidin MicroBeads, interacting partners such as activators or repressors can be magnetically pulled out of a cell lysate. Display technologies allow the discovery and characterization of new protein-protein interactions. μMACS Streptavidin MicroBeads can be used in combination with biotinylated target molecules (e.g. ligands) to screen a phage display library in order to identify interacting partners, for example receptors.⁷

Isolation of nucleic acids such as specific transcripts, viral sequences, and more
Specific nucleic acids can be detected and isolated using the biotinylated complementary sequence, for example, as single-stranded oligonucleotides. Thereby, catalytically active nucleic acids like ribozymes or viral sequences can be determined in bodily fluids. Furthermore, isolating specific transcripts that function as gene markers can be used to monitor the progression of a disease, such as an infection, or to identify stages of developmental differentiation. Further applications make use of μMACS Streptavidin MicroBeads for subtractive hybridization and serial analysis of gene expression (SAGE).

Viable HIV-1 virus isolation
Beside molecular isolation, μMACS Streptavidin MicroBeads are successfully used to isolate virions such as HIV-1. The gentle procedure yields viable and infectious virus from plasma, serum, or other bodily fluids without virion-destroying processing steps.⁸

Columns

μ Columns (included in kit)

Further information

Figure 1

Isolation of specific RNA binding proteins. Yeast extract was incubated with a full-length Mating Factor A2 mRNA bound to a 3'-biotinylated complementary ss-oligo and magnetically labeled with μMACS Streptavidin MicroBeads. As a control a magnetically labeled mutant mRNA, missing the binding site for Mating Factor A2 binding proteins was used. The figure shows the silver-stained SDS gel. Four proteins with apparent molecular weights of 33, 44, 48 and 51 kDa were isolated, which bind specifically to the RNA sequence; in the control experiment with the mutated RNA sequence no specific proteins were isolated. (Courtesy of Dr. Allan Albig, Washington State University, U.S.A.).

A Full-length RNA

B Mutant RNA