Streptavidin magnetic beads

Manufactured by BioLegend

Streptavidin magnetic beads are a type of laboratory equipment used for the separation and purification of biotinylated molecules. Streptavidin, a protein with a high affinity for biotin, is covalently attached to the surface of the magnetic beads, allowing for the efficient capture and isolation of biotin-labeled targets.

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Lab products found in correlation

Anti nk1.1 biotin, by BioLegend (1 mentions) Dnase 1, by Merck Group (1 mentions) Biotin conjugated anti cd11b ab, by BioLegend (1 mentions) Bx40 microscope, by Olympus (1 mentions) Infinity lite b camera, by Olympus (1 mentions) Iscript cdna synthesis kit, by Bio-Rad (1 mentions) Cfx connect instrument, by Bio-Rad (1 mentions) Nucleospin rna kit, by Macherey-Nagel (1 mentions) Iq sybr green supermix reagent, by Bio-Rad (1 mentions)

Spelling variants of this product

Magnetic beads (9 citations) Streptavidin conjugated magnetic beads (2 citations)

4 protocols using streptavidin magnetic beads

Cytotoxicity Assay for Tumor Immunity

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Cytotoxicity by splenocytes was assessed with a standard 4-hour ⁵¹Cr-release assay. ~24 hours after CDN or PBS treatment of tumors, spleens were harvested and treated with ACK lysing buffer. Pooled splenocytes from 4–6 mice were employed as effector cells. Triplicate samples of 10^{4 51}Cr-labeled RMA-B2m^−/− cells per 96-well V-bottom plate well were incubated with splenocytes at the indicated E:T ratios for 4 hrs before determining the percent ⁵¹Cr release in the supernatant. % specific lysis = 100 x (experimental - spontaneous release_Avg)/(maximum release_Avg - spontaneous release_Avg), where maximum release was release with addition of Triton X-100 (final concentration 2.5%).
Where shown, pooled splenocytes were NK-depleted by incubating on ice for 30 min with anti-NKp46-biotin (Biolegend) and anti-NK1.1-biotin (Biolegend) followed by 20 min incubation with streptavidin magnetic beads (Biolegend), and magnetic removal of bead-bound cells. Depletion (>95%) was confirmed by flow cytometry.

Nicolai C.J., Wolf N., Chang I.C., Kirn G., Marcus A., Ndubaku C.O., McWhirter S.M, & Raulet D.H. (2020). NK cells mediate clearance of CD8+ T cell–resistant tumors in response to STING agonists. Science immunology, 5(45), eaaz2738.

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Tumor-Associated Macrophage Isolation

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Under deep ether anesthesia, the liver was perfused with Hanks Balanced Salt Solution (HBSS) via the portal vein. Immediately after perfusion, the liver was removed. Tumors were separated from the liver (with a margin of approximately 2 mm from the tumor edge) and minced with scissors. Liver specimens were incubated in 10 ml of HBSS containing 0.05% Type IV collagenase and 0.01 mg/ml DNase I (Sigma). The specimens were shaken for 40 min at 37 °C and then filtered through a stainless-steel mesh (70 µm), suspended in 33% Percoll solution and centrifuged for 15 min at 450 g at room temperature. After the red blood cells were lysed, the remaining cells were washed twice^{38 (link),39 (link)}.
For TAM magnetic sorting, single cells prepared as described above were incubated with Fc-blocker and then stained with biotin-conjugated anti-F4/80 Ab (ebioscience) and incubated with streptavidin magnetic beads (Biolegend). Positively labeled cells were collected using a magnet. For monocyte/macrophage magnetic sorting during the monocyte maturation process, biotin-conjugated anti-CD11b Ab (Biolegend) was used, followed by the above-described steps.

Li M., Lai X., Zhao Y., Zhang Y., Li M., Li D., Kong J., Zhang Y., Jing P., Li H., Qin H., Shen L., Yao L., Li J., Dou K, & Zhang J. (2018). Loss of NDRG2 in liver microenvironment inhibits cancer liver metastasis by regulating tumor associate macrophages polarization. Cell Death & Disease, 9(2), 248.

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Sickled Cell Disease Morphology Analysis

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SCD cells at day 21 of differentiation were stained with CD71-biotin antibody (Invitrogen, catalog 13-0711-82, clone R17217; 1:200). CD71⁻ cells were isolated using streptavidin magnetic beads (BioLegend, catalog no. 480016). Cells were then resuspended in 100 μl of HEMOX buffer supplemented with 20 mM glucose and 0.32% bovine serum albumin (BSA). Cells were subsequently incubated under 2.5% O₂ at 37 °C for 1 h. Cells were immediately fixed with 200 μl of 2% glutaraldehyde solution. Fixed-cell suspensions were spread on to glass microslides and subjected to microscopic morphological analysis of bright-field images of single-layer cells on an Olympus BX40 microscope fitted with an Infinity Lite B camera (Olympus). The cells with irregular shapes (sickled shape or protruding structures) were counted as sickled cells. Cell shapes were measured by software CellProfiler⁵⁰. Irregular shape was determined by the cell area compactness (cutoff 1.3). Over 1,300 cells were analyzed for each condition.

Huang P., Peslak S.A., Ren R., Khandros E., Qin K., Keller C.A., Giardine B., Bell H.W., Lan X., Sharma M., Horton J.R., Abdulmalik O., Chou S.T., Shi J., Crossley M., Hardison R.C., Cheng X, & Blobel G.A. (2022). HIC2 controls developmental hemoglobin switching by repressing BCL11A transcription. Nature genetics, 54(9), 1417-1426.

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Isolation and Analysis of CD8+ T cells

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For gene-expression analysis, tumor-infiltrating CD8 + T cells were isolated from enriched tumor-infiltrating lymphocytes by incubating lymphocytes in biotin anti-CD8a (BioLegend) followed by streptavidin magnetic beads (BioLegend). The purity of isolated CD8 + T cells exceeds 95 %. Total RNA from cells were extracted using the NucleoSpin® RNA kit (Macherey-Nagel) according to the manufacturer's protocol. Reverse-transcription of the RNA was performed using random primers iScript™ cDNA Synthesis Kit (Bio-Rad). Realtime PCR and data collection were performed with IQ SYBR Green supermix reagent (Bio-Rad) on a CFX connect instrument (Bio-Rad) and the following primers: β-actin forward, 5´-AGCCATGTACGTAGCCATCC-3´; β-actin reverse, 5´-TCTCAGCTGTGGTGGTGAAG-3´; IFN-γ forward, 5´-CAGCAACAGCAAGGCGAAAA-3´; IFN-γ reverse, 5´-TCATTGAATGCTTGGCGCTG-3´; TNF-α forward, 5´-ATGGCCTCCCTCTCATCAGT-3´; TNF-α reverse, 5´-TTTGCTACGACGTGGGCTAC-3´. Human Furin, PCAE4, PC5/6, PC7
and GAPDH primers will be provided upon request. Each sample was run in triplicate and normalized to the housekeeping gene GAPDH. Relative gene expression was calculated using the ∆CT method with the equation 2∆∆CT. The results are shown as fold changes compared to the Furin expression or control group.

He Z., Khatib A.M, & Creemers J.W.M. (2020). Loss of the proprotein convertase Furin in T cells represses mammary tumorigenesis in oncogene-driven triple negative breast cancer. Cancer letters, 484.

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