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Streptavidin conjugated magnetic beads

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Streptavidin-conjugated magnetic beads are a type of laboratory equipment used for various biomedical applications. Streptavidin is a protein that binds strongly to the small molecule biotin, and these beads leverage this interaction to capture and isolate biotinylated molecules, proteins, or cells from complex samples. The magnetic properties of the beads allow for easy separation and manipulation using a magnetic field.

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Lab products found in correlation

Rneasy mini kit, by Qiagen (5 mentions) Rnase free dnase 1, by Promega (3 mentions) T7 rna polymerase, by Promega (3 mentions) Trizol reagent, by Thermo Fisher Scientific (2 mentions) Biotinylated peptide, by GenScript (1 mentions) Scaffold software, by Proteome Software (1 mentions) Agilent 2100 bioanalyzer, by Agilent Technologies (1 mentions) Complete edta free protease inhibitor, by Roche (1 mentions) Absolutely rna nanoprep kit, by Agilent Technologies (1 mentions) Rnasin, by Promega (1 mentions) Nanodrop 1000 spectrophotometer, by Agilent Technologies (1 mentions) Lipofectamine 2000 reagent, by Thermo Fisher Scientific (1 mentions) Bio nc, by Sangon (1 mentions) Lipofectamine rnaimax, by Thermo Fisher Scientific (1 mentions) Lysis buffer, by Thermo Fisher Scientific (1 mentions) Ez link sulfo nhs lc biotin, by Thermo Fisher Scientific (1 mentions) Megascript t7 transcription kit, by Thermo Fisher Scientific (1 mentions) Pierce biotin 3 end dna labeling kit, by Thermo Fisher Scientific (1 mentions) Dnase 1, by Merck Group (1 mentions) Magnetic column, by Miltenyi Biotec (1 mentions)

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Streptavidin magnetic beads (382 citations) Magnetic beads (264 citations) Streptavidin beads (263 citations) Streptavidin-conjugated beads (12 citations)

34 protocols using streptavidin conjugated magnetic beads

1

Constructing Probe-coated Magnetic Beads

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Probe-coated beads were constructed by incubating biotin-coupled circCYP24A1 or oligo probes with streptavidin-conjugated magnetic beads (Life Technologies, USA) for 3 h at 25 °C, which were used for RNA-pulldown analysis described in a previous study [23 (link)].
Wu X., Zhou J., Zhao L., Yang Z., Yang C., Chen Y, & Xue W. (2022). CircCYP24A1 hampered malignant phenotype of renal cancer carcinoma through modulating CMTM-4 expression via sponging miR-421. Cell Death & Disease, 13(2), 190.
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2

Pull-down analysis of Dectin-1 and CD69 interactions

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For pull-down assays, C57BL/6 mice were subcutaneously injected with Flt3 ligand-expressing B16 melanoma cells in order to expand the pool of dendritic cells (DC) in the spleen (27 (link)). After 12 days, CD11c+ cells were isolated and total lysates were prepared in lysis buffer (TBS - 1% NP40). Soluble lysates or, alternatively, recombinant tSHIP-1 (28 (link)) were incubated with biotinylated peptides (GenScript, Piscataway, NJ) corresponding to the cytoplasmic tail of mouse Dectin-1 (NH2-MKYHSHIENLDEDGYTQLDFSTQDIHKR-C, biotin in C-terminal lysine), phosphorylated or not in the Tyr3 and 15, or mouse CD69 (NH2-MDSENCSITENSSSHLERGQKDHGTSIHFEK-C, biotin in C-terminal lysine). Peptides were pulled down using streptavidin-conjugated magnetic beads (Life Technologies, Carlsbad, CA) and interacting proteins were analyzed by mass spectrometry or western blotting using antibodies against SYK (N-1), SHIP-1 (P1C1) for Fig. 1C and Supplemental Fig. 1B or (D20 and V19) for Fig. 1D (Santa Cruz, Dallas, TX), LYN (C13F9) and SHP-2 (#3752) (Cell Signaling). Scaffold software (Proteome software, Runcorn, UK) was used to analyze mass spectrometry data.
Blanco-Menéndez N., del Fresno C., Fernandes S., Calvo E., Conde-Garrosa R., Kerr W.G, & Sancho D. (2015). SHIP-1 couples to the Dectin-1 hemITAM and selectively modulates reactive oxygen species production in dendritic cells in response to C. albicans. Journal of immunology (Baltimore, Md. : 1950), 195(9), 4466-4478.
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3

Affinity Purification of EGFP-Tagged Polysomes

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Affinity purification of EGFP-tagged polysomes was done 3–4 weeks after virus injections. Three biological replicates consisting of tissue pooled from 3–5 mice (mixed sex) were collected for each condition. IPs and RNA extractions were carried out as previously described (Heiman et al., 2014 (link)) and in Supplemental Experimental Procedures. Briefly, brain tissue was homogenized in buffer containing 10 mM HEPES-KOH (pH 7.4), 150 mM KCl, 5 mM MgCl2, 0.5 mM DTT, 100 μg/ml cycloheximide, RNasin (Promega, Madison, WI) and SUPERas-InTM (Life Technologies) RNase inhibitors, and Complete-EDTA-free protease inhibitors (Roche), and then cleared by two-step centrifugation to isolate polysome-containing cytoplasmic supernatant. Polysomes were immunoprecipitated using monoclonal anti-EGFP antibodies (clones 19C8 and 19F7; see Heiman et al., 2008 (link)) bound to biotinylated-Protein L (Pierce, Thermo Fisher, Waltham, MA) coated streptavidin-conjugated magnetic beads (Life Technologies) and bound RNA was purified using the Absolutely RNA Nanoprep kit (Agilent). RNA quantity was measured with a Nanodrop 1000 spectrophotometer and quality was assayed on an Agilent 2100 Bioanalyzer. Only samples with RNA integrity values ≥7.0 were used for RNA-seq and qRT-PCR analyses.
Nectow A.R., Moya M.V., Ekstrand M.I., Mousa A., McGuire K.L., Sferrazza C.E., Field B.C., Rabinowitz G.S., Sawicka K., Liang Y., Friedman J.M., Heintz N, & Schmidt E.F. (2017). Rapid Molecular Profiling of Defined Cell Types Using Viral TRAP. Cell reports, 19(3), 655-667.
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4

Biotin-tagged miRNA Pulldown Assay

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Biotin-conjugated miR-657 (50 nM), Biotin-conjugated miR-657 antisense (50 nM), miR-657 mimics (50 nM), miR-657 antisense (50 nM) were transfected into A549 and HCC827 cells using Lipofectamine™ 2000 reagent (Invitrogen) and cultured for 48 h. After that, cells were harvested (1 × 107 cells) and the cells were lysed, then500 μl of the lysates were incubated with 500 μl of streptavidin-conjugated magnetic beads (Life Technologies) at room temperature for 2 h. After washing, the RNA was extracted with TRIzol and evaluated by PCR.
Zhang Y., Yuan J., Guo M., Xiang R., Xie T., Zhuang X., Dai W., Li Q, & Lai Q. (2023). Upregulation of long intergenic non-coding RNA LINC00326 inhibits non-small cell lung carcinoma progression by blocking Wnt/β-catenin pathway through modulating the miR-657/dickkopf WNT signaling pathway inhibitor 2 axis. Biology Direct, 18, 3.
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5

Investigating circKRT17-EIF4A3 Interaction

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An RNA pull-down assay was utilized to determine the interaction between circKRT17 and EIF4A3 using biotin-labeled circKRT17. Biotin-labeled circKRT17 was designed and provided by GenePharm (Shanghai, China). A specific circKRT17 probe was mixed and incubated with cell lysates of the indicated LUAD cells overnight. Next, streptavidin-conjugated magnetic beads (Invitrogen, Carlsbad, CA, USA) were added and incubated for another 2 h. Beads were then rinsed and interested protein levels were detected via western blotting.
Ji Y., Zhao Q., Feng W., Peng Y., Hu B, & Chen Q. (2022). N6-Methyladenosine Modification of CIRCKRT17 Initiated by METTL3 Promotes Osimertinib Resistance of Lung Adenocarcinoma by EIF4A3 to Enhance YAP1 Stability. Cancers, 14(22), 5582.
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6

Identifying circ-PGAP3 and miR-330-3p Targets

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MDA-MB-231 and HS598T cell lysates were collected and incubated with biotin-labeled circ-PGAP3 probe, or biotin-labeled miR-330-3p mimics were transfected into TNBC cells. After that, cell lysates were incubated with streptavidin-conjugated magnetic beads (Invitrogen) at 25 °C for 3 h. After the beads were attached to the magnetic frame, the lysates were discarded and Trizol was added to extract total RNA, followed by qRT-PCR analysis of circ-PGAP3 or miRNA enrichment.
He D., Yang X., Kuang W., Huang G., Liu X, & Zhang Y. (2020). The Novel Circular RNA Circ-PGAP3 Promotes the Proliferation and Invasion of Triple Negative Breast Cancer by Regulating the miR-330-3p/Myc Axis. OncoTargets and therapy, 13, 10149-10159.
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7

Affinity Purification of lincRNA Protein Interactors

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Biotin-labelled lincRNA-MIR99AHG were transcribed with biotin RNA labelling mix and T7 RNA polymerase (Promega), treated with RNase-free DNase I (Promega), and purified using RNeasy Mini Kit (Qiagen). Biotin-labeled lincRNA-MIR99AHG was mixed and incubated with nuclear extracts of IL-4/IL-13 stimulated and Mtb HN878 infected BMDMs. Streptavidin-conjugated magnetic beads (Invitrogen) were added to each binding reaction and further incubated. Beads were washed thoroughly. Retrieved proteins were detected by mass spectrometry.
Gcanga L., Tamgue O., Ozturk M., Pillay S., Jacobs R., Chia J.E., Mbandi S.K., Davids M., Dheda K., Schmeier S., Alam T., Roy S., Suzuki H., Brombacher F, & Guler R. (2022). Host-Directed Targeting of LincRNA-MIR99AHG Suppresses Intracellular Growth of Mycobacterium tuberculosis. Nucleic Acid Therapeutics, 32(5), 421-437.
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8

Investigating PVT1-miR-124 Interaction in HK-2 Cells

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RNA pull down assay was carried out to test possible interaction between PVT1 and miR-124 in HK-2 cells as described previously.39 (link) Biotin-labeled wide type miR-124 (Bio-miR-124-WT), biotin-labeled mutant type miR-124 (Bio-miR-124-MUT) with mutant PVT binding sites, biotin-labeled wide type PVT1 probe (Bio-probe-PVT1-WT), biotin-labeled mutant type PVT1 probe containing mutant miR-124 binding sites (Bio-probe-PVT1-MUT) and a biotin-labeled control probe (Bio-NC) were synthesized by Sangon Inc. (Shanghai, China). Then, biotin-labeled probes or miRNAs were incubated with streptavidin-conjugated magnetic beads (Invitrogen) and added into HK-2 cell lysate. Finally, the enrichment degrees of PVT1 and miR-124 in eluted RNA complex were determined by RT-qPCR assay.
Zhu X., Shi J., li H, & Chen F. (2018). Retracted Article: PVT1 knockdown alleviates vancomycin-induced acute kidney injury by targeting miR-124 via inactivation of NF-κB signaling. RSC Advances, 8(55), 31725-31734.
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9

Quantifying Protein-Lipid Interactions with Biotinylated Liposomes

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For most experiments, 0.5 µg of purified GST fusion protein was incubated with 3 µM biotinylated liposomes composed of 5% PIP lipids in 20 mM Hepes, 120 mM NaCl, 1 mM EGTA, 0.2 mM CaCl2, 1 mM MgCl2, 5 mM KCl, and 1 mg/ml fatty acid-free BSA, pH 7.4, for 15 min at room temperature. For the binding isotherm (Fig. 1 D) and competition assays (Fig. 5, E and F), 70 nM cortactin was used instead of 0.5 µg. Liposome–protein complexes were then further incubated with streptavidin-conjugated magnetic beads (Invitrogen) for 30 min at room temperature. After washing with binding buffer and isolation with a magnet, liposome-bound protein was analyzed by SDS-PAGE. The intensity of bands was quantified by ImageJ software (NIH). To compare relative binding affinity of each protein to PI(3,5)P2, nonspecific bead binding band intensity (beads only lane) was subtracted from the specific liposome-bead band intensity, and then normalized by the intensity of the band in the input lane. Apparent equilibrium Kd were calculated using a one-site binding hyperbola nonlinear regression analysis in Prism (GraphPad, version 5 software).
Hong N.H., Qi A, & Weaver A.M. (2015). PI(3,5)P2 controls endosomal branched actin dynamics by regulating cortactin–actin interactions. The Journal of Cell Biology, 210(5), 753-769.
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10

Biotinylated miR-326 Mimics Pulldown

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Using Lipofectamine RNAiMax (Life Technologies), OA-FLS were transfected with biotinylated miR-326 mimics or mutant (RiboBio, Guangzhou, China, 50 nM). After 48 hours, cells were incubated with a lysis buffer (Ambion). The lysates were further incubated with streptavidin-conjugated magnetic beads (Invitrogen, United States). 10% of the cell lysates were aliquot for input. RNAs bound to the beads were extracted using the RNeasy Mini Kit (QIAGEN) for the subsequent analysis.
Lin Z., Ma Y., Zhu X., Dai S., Sun W., Li W., Niu S., Chu M, & Zhang J. (2022). Potential predictive and therapeutic applications of small extracellular vesicles-derived circPARD3B in osteoarthritis. Frontiers in Pharmacology, 13, 968776.
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