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M 280 streptavidin coated magnetic beads

Manufactured by Thermo Fisher Scientific
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The M-280 streptavidin-coated magnetic beads are a versatile laboratory tool designed for various biomolecular separation and purification applications. Streptavidin, a high-affinity protein, is covalently coupled to the surface of these superparamagnetic beads, enabling efficient capture and separation of biotinylated molecules, such as proteins, nucleic acids, and cells. The magnetic properties of these beads allow for easy manipulation and separation using a suitable magnetic separator.

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Lab products found in correlation

Ampligase, by Illumina (2 mentions) Trizol reagent, by Thermo Fisher Scientific (2 mentions) Pierce magnetic rna protein pull down kit, by Thermo Fisher Scientific (2 mentions) Protease inhibitor, by Solarbio (2 mentions) Lysis buffer, by Promega (1 mentions) Human il 1β, by BioLegend (1 mentions) Sars cov 2 rbd mfc protein, by Sino Biological (1 mentions)

9 protocols using m 280 streptavidin coated magnetic beads

1

DNA Ligation and Capture Protocol

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Example 2

A protocol suitable for performing the method illustrated in FIG. 2 is as follows: 1) 10 ng of DNA is digested with 1 unit of restriction enzyme in corresponding compatible restriction enzyme buffer. The reaction is incubated in 37 C for 1 h, followed by enzymatic deactivation at 80 C for 20 min. 2) The DNA fragments are denatured to single stranded fragments at 95 C for 10 min and mixed with probes and ligase to form linear ligation products. The probe pool are added in 10 pM individual concentration along with 1 U of Ampligase (Epicentre) and incubated at 55 C for 1 h in ligase buffer. 3) The ligation product is captured on magnetic streptavidin beads. To remove non-reacted probes and fragments, the solution is mixed with 10 ml M-280 streptavidin coated magnetic beads (Invitrogen) in Tris-HCl (pH 7.5), 3.5 mM EDTA and 0.07% Tween-20 in a final volume of 200 ml, and incubated at room temperature for 15 min. After incubation, the beads are collected using a ring magnet and supernatant removed.

US10731214B2. Nucleic acid probe and method of detecting genomic fragments (2020-08-04). Vanadis Diagnostics [SE]. Inventors: Carl Oscar Fredrik Dahl [SE], Olof John Ericsson [SE].
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2

DNA Ligation Protocol for Targeted Sequencing

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Example 2

A protocol suitable for performing the method illustrated in FIG. 2 is as follows: 1) 10 ng of DNA is digested with 1 unit of restriction enzyme in corresponding compatible restriction enzyme buffer. The reaction is incubated in 37 C for 1 h, followed by enzymatic deactivation at 80 C for 20 min. 2) The DNA fragments are denatured to single stranded fragments at 95 C for 10 min and mixed with probes and ligase to form linear ligation products. The probe pool are added in 10 pM individual concentration along with 1 U of Ampligase (Epicentre) and incubated at 55 C for 1 h in ligase buffer. 3) The ligation product is captured on magnetic streptavidin beads. To remove non-reacted probes and fragments, the solution is mixed with 10 ml M-280 streptavidin coated magnetic beads (Invitrogen) in Tris-HCl (pH 7.5), 3.5 mM EDTA and 0.07% Tween-20 in a final volume of 200 ml, and incubated at room temperature for 15 min. After incubation, the beads are collected using a ring magnet and supenatant removed.

US10240198B2. Nucleic acid probe and method of detecting genomic fragments (2019-03-26). Vanadis Diagnostics [SE]. Inventors: Carl Oscar Fredrik Dahl [SE], Olof John Ericsson [SE].
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3

Detecting lncRNA H19-miR-130a Interaction

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RNA pull down was performed to detect the interplay between lncRNA H19 and miR-130a. In brief, C28/I2 cells were harvested and rinsed with cold PBS solution three times, followed by the addition of 1 mL of lysis buffer containing protease inhibitor (Solarbio, Beijing, China). Cell lysates were mixed with biotin-labeled miRNAs or lncRNA probe sequences, and incubated with M-280 streptavidincoated magnetic beads (Invitrogen) for 30 min at room temperature. Then, the beads were washed with ice-cold lysis buffer, low-salt buffer, and high-salt buffer, in this order, and bound RNAs was extracted using TRIzol reagent (Invitrogen) and quantified by RT-qPCR assay.
Hu Y., Li S, & Zou Y. (2019). Knockdown of LncRNA H19 Relieves LPS-Induced Damage by Modulating miR-130a in Osteoarthritis. Yonsei Medical Journal, 60(4), 381-388.
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4

Detecting PVT1-miR-24-3p Interaction

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RNA pull-down was allowed to determine the interaction between PVT1 and miR-24-3p. HEK293 cells or RN-c cells were added with lysis buffer containing protease inhibitors (Solarbio, Beijing, China), mixed with biotin-labeled lncRNA probe sequence, and incubated with M-280 streptavidin-coated magnetic beads (Invitrogen). After washing the beads with ice-cold lysis buffer, low-salt buffer and high-salt buffer in sequence, the bound RNA was collected using Trizol reagent (Invitrogen), and quantified by RT-qPCR.
Chen Z., Fan T., Zhao X, & Zhang Z. (2021). Depleting SOX2 improves ischemic stroke via lncRNA PVT1/microRNA-24-3p/STAT3 axis. Molecular Medicine, 27, 107.
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5

DANCR-miR-34a-5p Interaction Examination

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The interaction between DANCR and miR-34a-5p was examined using the Pierce Magnetic RNA-Protein Pull-Down Kit (Thermo Fisher Scientific) in line with the manufacturer’s protocols. Biotin-labeled wild-type DANCR (Bio-DANCR WT) containing the putative miR-34a-5p binding sites and the biotin-labeled mutant DANCR (Bio-DANCR MUT) designed to disrupt the base-pairing between DANCR and miR-34a-5p were mixed with protein extracts of DU145/DTX and PC3/DTX cells and M-280 streptavidin-coated magnetic beads (Invitrogen). The beads were subsequently washed with ice-cold lysis buffer, low-salt buffer, and high-salt buffer. Following that, bound RNAs were purified from the RNA–protein complex and miR-34a-5p expression was determined by qRT-PCR.
Ma Y., Fan B., Ren Z., Liu B, & Wang Y. (2019). Long noncoding RNA DANCR contributes to docetaxel resistance in prostate cancer through targeting the miR-34a-5p/JAG1 pathway. OncoTargets and therapy, 12, 5485-5497.
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6

Biotinylated miR-149 Pulldown Assay

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Chondrocytes were transfected with 50 nM biotinylated miR-149 or the mut (GenePharma) for 48 h. After rinsing with cold PBS, cells were lysed with lysis buffer (Promega). Afterward, one half of specimen was aliquoted for input. The other samples were incubated with M-280 streptavidin-coated magnetic beads (Invitrogen) at 4°C. Three hours later, the samples were washed twice with ice-cold lysis buffer and subsequent rinsed three times with low-salt buffer. Then, the specimens were rinsed with high-salt buffer containing 0.1% SDS, 2 mM 193 EDTA, 1% Trition X-100, 500 mM NaCl, 20 mM Tris-HCl with pH 8.0. The bound RNAs were then extracted using TRIzol reagent and subjected into qRT-PCR assay to detect lncRNA PVT1 enrichment.
Zhao Y., Zhao J., Guo X., She J, & Liu Y. (2018). Long non-coding RNA PVT1, a molecular sponge for miR-149, contributes aberrant metabolic dysfunction and inflammation in IL-1β-simulated osteoarthritic chondrocytes. Bioscience Reports, 38(5), BSR20180576.
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7

Probing lncRNA-H19 and miR-29b-3p Interaction

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The interaction between lncRNA-H19 and miR-29b-3p was examined using the Pierce Magnetic RNA-Protein Pull-Down Kit (Thermo Fisher Scientific) according to the manufacturer's protocols. Biotin-labeled wild-type lncRNA-H19 (Bio-H19 WT) containing the putative miR-29b-3p binding sites and the biotin-labeled mutant lncRNA-H19 (Bio-H19 MUT) designed to disrupt the base pairing between lncRNA-H19 and miR-29b-3p were mixed with protein extracts of SKOV3-CB cells and M-280 streptavidin-coated magnetic beads (Invitrogen). The beads were subsequently washed with ice-cold lysis buffer, low-salt buffer, and high-salt buffer. Next, bound RNAs were purified from the RNA-protein complex, and miR-29b-3p expression was determined by qRT-PCR.
Tian X., Zuo X., Hou M., Li C, & Teng Y. (2021). LncRNA-H19 regulates chemoresistance to carboplatin in epithelial ovarian cancer through microRNA-29b-3p and STAT3. Journal of Cancer, 12(19), 5712-5722.
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8

Isolation of Anti-IL-1β Antibodies

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Anti-IL-1β antibodies were isolated from HX02 human Fab phage display library (Humanyx Pte Ltd) via in vitro selection. We followed the procedures of biopanning, phage amplification, Fab expression and purification described by de Haard et al.39 (link) Briefly, biopanning was performed using human IL-1β (Biolegend) biotinylated using the EZ-Link NHS-PEG4-Biotin labeling kit (Pierce). In the first two rounds of biopanning, IL-1β was immobilized on M280 streptavidin-coated magnetic beads (Life Technologies); in the third round, biotinylated IL-1β was immobilized on the neutravidin-coated microplate in order to avoid isolation of streptavidin magnetic beads binders. 1013 cfu phage in 1 mL casein-PBS blocking buffer was used in the first round, and 1011 cfu phage were used in the second and third rounds. After three rounds of biopanning, the Fab of selected clones was expressed in E. coli TG1 cells (Stratagene) to screen for IL-1β binders by ELISA. Unique clones were identified by DNA fingerprinting technique as previously described39 (link) and confirmed by DNA sequencing.
Goh A.X., Bertin-Maghit S., Ping Yeo S., Ho A., Derks H., Mortellaro A, & Wang C.I. (2014). A novel human anti-interleukin-1β neutralizing monoclonal antibody showing in vivo efficacy. mAbs, 6(3), 764-772.
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9

Isolation of Anti-SARS-CoV-2 Spike RBD Antibodies

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Anti-SARS-CoV-2 Spike RBD antibodies were isolated from an HX02 human Fab phage display library (Humanyx Pte Ltd) via in vitro selection. Briefly, biopanning was performed using SARS-CoV-2 RBD (YP_009724390.1) (Arg319-Phe541) with a mouse Fc tag (Sino Biological, Cat#40592-V05H) biotinylated using the EZ-Link NHS-PEG4-Biotin labeling kit (Thermo Fisher Scientific, Cat#A39259). In both rounds of biopanning, biotinylated SARS-CoV-2 RBD-mFc protein (Sino Biological, Cat#40592-V05H) was immobilized on M280 streptavidin-coated magnetic beads (Life Technologies, Cat#11205D); 3.5 × 1012 cfu phage in 1ml 1% casein-PBS blocking buffer was used in the first round, and 1.64 × 1011 cfu phage were used in the second round. During the biopanning process, binders to mouse Fc were removed by pre-incubation of phage with 2 μM mouse IgG before mixing with the RBD-mFc antigen. After two rounds of biopanning, the Fabs of selected clones were expressed in E. coli HB2151 cells (Stratagene) to screen for RBD binders by ELISA. Unique clones were identified by DNA sequencing.
Asarnow D., Wang B., Lee W.H., Hu Y., Huang C.W., Faust B., Ng P.M., Ngoh E.Z., Bohn M., Bulkley D., Pizzorno A., Ary B., Tan H.C., Lee C.Y., Minhat R.A., Terrier O., Soh M.K., Teo F.J., Yeap Y.Y., Seah S.G., Chan C.E., Connelly E., Young N.J., Maurer-Stroh S., Renia L., Hanson B.J., Rosa-Calatrava M., Manglik A., Cheng Y., Craik C.S, & Wang C.I. (2021). Structural insight into SARS-CoV-2 neutralizing antibodies and modulation of syncytia. Cell, 184(12), 3192-3204.e16.
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