Streptavidin conjugated microbeads

Manufactured by Miltenyi Biotec

Sourced in Germany

Streptavidin-conjugated microbeads are a type of lab equipment used for the isolation and purification of biotinylated molecules. Streptavidin, a protein with a high affinity for biotin, is covalently attached to the surface of the microbeads, allowing for the efficient capture of biotinylated targets.

Automatically generated - may contain errors

Lab products found in correlation

Facsaria sorter, by BD (4 mentions) Automacs system, by Miltenyi Biotec (3 mentions) Recombinant murine tnf, by Thermo Fisher Scientific (2 mentions) Recombinant cd70 fc, by Sino Biological (2 mentions) Soluble cd70, by Enzo Life Sciences (2 mentions) Agonist cd27 mab rm27 3e5, by R&D Systems (2 mentions) Recombinant mouse cd30 l and 4 1bbl, by R&D Systems (2 mentions) Anti poly his antibody, by R&D Systems (2 mentions) Rs 2000 biological irradiator, by Rad Source (2 mentions) Human il 2, by R&D Systems (2 mentions) Biotinylated anti cd8 and ter119 antibodies, by Thermo Fisher Scientific (2 mentions) Mouse il4, by Cytek Biosciences (2 mentions) Rneasy kit, by Qiagen (2 mentions) Aria sorter, by BD (2 mentions) Collagenase 4, by Merck Group (1 mentions) Ls column, by Miltenyi Biotec (1 mentions) Anti cd3, by Thermo Fisher Scientific (1 mentions) Anti cd28, by Thermo Fisher Scientific (1 mentions) Facscanto, by BD (1 mentions) Sh800 cell sorter, by Sony (1 mentions)

Close spelling variants of this product

Streptavidin microbeads (158 citations) MACS streptavidin conjugated microbeads (2 citations) Streptavidin-conjugated magnetic microbeads (2 citations)

24 protocols using streptavidin conjugated microbeads

Isolation of anti-TEM8 antibody m830

Cited 1 time

Check if the same lab product or an alternative is used in the 5 most similar protocols

The m830 antibody was derived from a yeast display library constructed using a collection of human antibody gene repertoires^{69 (link)} and those from more than 50 additional individuals. Due to the in vitro stochastic pairing of VH and VL repertoires, this library is not subject to tolerance mechanisms found in normal immune responses and allowed the generation of antibodies against regions of the TEM8 ECD that are 100% conserved between mouse and human. In vitro selection of the yeast display library involved multiple rounds of sequential panning on biotinylated, purified recombinant human or murine TEM8(ED)-AP and TEM8-Fc fusion proteins. Biotinylated recombinant TEM8 proteins were incubated with 5 × 10¹⁰ cells from the antibody library in PBS-BSA (PBS containing 0.1% BSA) for 2 h, washed with PBS-BSA, and captured with streptavidin-conjugated microbeads from Miltenyi Biotec using the AutoMACS System. The sorted cells were amplified, and the panning was repeated. After further scFv validation, m830, one of the lead antibodies from this screen, was converted into a full-size human IgG1. m830 was collected from culture supernatants grown in serum free medium and purified by protein A chromatography. Antibody preparations for in vivo studies possessed less than 5% aggregates and had endotoxin levels below 1 EU/mg.

Hsu K.S., Dunleavey J.M., Szot C., Yang L., Hilton M.B., Morris K., Seaman S., Feng Y., Lutz E.M., Koogle R., Tomassoni-Ardori F., Saha S., Zhang X.M., Zudaire E., Bajgain P., Rose J., Zhu Z., Dimitrov D.S., Cuttitta F., Emenaker N.J., Tessarollo L, & St. Croix B. (2022). Cancer cell survival depends on collagen uptake into tumor-associated stroma. Nature Communications, 13, 7078.

+ Open protocol

+ Expand

CNS Mononuclear Cell Isolation

Cited 1 time

Check if the same lab product or an alternative is used in the 5 most similar protocols

Brain and spinal cord tissues from mice perfused with PBS were digested with collagenase IV (Sigma-Aldrich) for 30 min at 37 °C, resuspended in 37 % Percoll, and loaded between a 30 and 70 % Percoll gradient. After centrifugation at 2000g for 20 min, CNS mononuclear cells were retrieved from the 37/70 % Percoll interface as previously done [38 (link)]. PMN- and MO-MDSC subsets were purified from CNS mononuclear cells by immunomagnetic separation using biotinylated anti-Ly6G and anti-Gr1 antibody and streptavidin-conjugated MicroBeads as mentioned above (Miltenyi Biotec). Cell purity was >95 % by flow cytometric analysis using anti-CD11b and Gr-1 antibodies.

Cantoni C., Cignarella F., Ghezzi L., Mikesell B., Bollman B., Berrien-Elliott M.M., Ireland A.R., Fehniger T.A., Wu G.F, & Piccio L. (2016). Mir-223 regulates the number and function of myeloid-derived suppressor cells in multiple sclerosis and experimental autoimmune encephalomyelitis. Acta neuropathologica, 133(1), 61-77.

+ Open protocol

+ Expand

Generating Anti-CD276 scFv Antibodies from Yeast Display Library

Cited 2 times

Check if the same lab product or an alternative is used in the 5 most similar protocols

A yeast display naïve, single-chain variable fragment (scFv) human antibody library was used to generate the anti-human CD276 scFvs as described previously (35 (link)). The library was constructed using a collection of human antibody gene repertoires, including the genes used for the construction of a phage display Fab library and those from more than 50 additional individuals and contained a total of 1e10 scFvs. In vitro selection of the yeast display library involved three rounds of sequential panning on biotinylated, purified recombinant CD276(ED)-AP (alkaline phosphatase) fusion proteins. For this, 10 μg of biotinylated hCD276(ED)-AP was incubated with approximately 5e10 cells from the initial naïve antibody library in 50 mL PBSA (PBS containing 0.1% BSA) for 2 hours, washed with PBSA, and captured with streptavidin-conjugated microbeads from Miltenyi Biotec using the AutoMACS system. The sorted cells were amplified and the panning was repeated once with the human hCD276 (ED)-AP and once with the mouse mCD276(ED)-AP protein to enrich for cross-reactive binders. After characterizing several scFvs for binding specificity, cross-species reactivity, a panel of five binders (1 (link)–5 (link)) were sequenced and used for CAR construction.

Majzner R.G., Theruvath J.L., Nellan A., Heitzeneder S., Cui Y., Mount C.W., Rietberg S.P., Linde M.H., Xu P., Rota C., Sotillo E., Labanieh L., Lee D.W., Orentas R.J., Dimitrov D.S., Zhu Z., St Croix B., Delaidelli A., Sekunova A., Bonvini E., Mitra S.S., Quezado M.M., Majeti R., Monje M., Sorensen P.H., Maris J.M, & Mackall C.L. (2019). CAR T Cells Targeting B7-H3, a Pan-Cancer Antigen, Demonstrate Potent Preclinical Activity Against Pediatric Solid Tumors and Brain Tumors. Clinical cancer research : an official journal of the American Association for Cancer Research, 25(8), 2560-2574.

+ Open protocol

+ Expand

Isolation and Characterization of Activated T-cells

Check if the same lab product or an alternative is used in the 5 most similar protocols

Cells cultured in vitro were harvested at various time-points, re-suspended in labeling buffer (2% FBS in PBS), and FAC-sorted a second time as CD4⁺ or CD8α⁺ CD69⁺ and either IL-2.eGFP⁺/Thy1.1⁺ or as IL-2.eGFP⁻/Thy1.1⁻. SMARTA IL-2.eGFP T-cells isolated ex vivo from acutely-activated recipient mice were processed from tissue by negative selection with biotinylated antibodies to CD11b, CD11c, and B220, streptavidin-conjugated microbeads, and LS columns (Miltenyi Biotech). Column flow-through fractions were then stained for a congenic marker (Thy1.1, Thy1.2, or CD45.2) prior to sorting as above.

DiToro D., Winstead C.J., Pham D., Witte S., Andargachew R., Singer J.R., Wilson C.G., Zindl C.L., Luther R.J., Silberger D.J., Weaver B.T., Kolawole E.M., Martinez R.J., Turner H., Hatton R.D., Moon J.J., Way S.S., Evavold B.D, & Weaver C.T. (2018). Differential IL-2 Expression Defines Developmental Fates of Follicular versus Non-follicular Helper T cells. Science (New York, N.Y.), 361(6407), eaao2933.

+ Open protocol

+ Expand

Isolation and Culture of Treg Progenitors

Check if the same lab product or an alternative is used in the 5 most similar protocols

T_reg progenitors were isolated as previously described^{9 (link)}. Briefly, 6-8 week old Foxp3-GFP thymii were dissected, dissociated, and pooled (up to 8 mice per experiment). CD4SP cells were enriched by magnetically depleting with biotinylated anti-CD8, CD11B, CD11C, CD19, CD45R/B220, MHC class II, NK1.1, and Ter119 antibodies (eBioscience) followed by secondary labeling with streptavidin-conjugated microbeads (Miltenyi Biotec). Enriched CD4SP cells were stained with fluorchrome-conjugated anti-CD4, CD25, and streptavidin prior to sorting CD4⁺CD25⁺GFP⁻ cells using a BD Facs Aria sorter (BD Biosciences). Purified T_reg progenitors were incubated in complete RPMI and supplemented with human IL-2 (1 Unit/mL, from the NIH repository at Frederick National Laboratory) with or without 100 nM recombinant GITR-L–Fc, OX40-L–Fc (both from Imgenex, San Diego, CA), recombinant murine TNF (Peprotech, Rocky Hill, NJ), recombinant CD70-Fc (Sino Biologicals, Beijing, China), soluble CD70 (Enzo Life Sciences, Farmingdale, NY), agonist CD27 mAb (RM27-3E5)^{47 (link)}, or recombinant mouse CD30-L and 4-1BBL (the latter two of which were crosslinked using 5 ug/ml anti-poly His antibody, both from R&D Systems, Minneapolis, MN). After 72 hours, cells were harvested, stained with anti-CD4 and CD25 antibodies and analyzed by flow cytometry for the percentage of cells that express GFP after incubation.

Mahmud S.A., Manlove L.S., Schmitz H.M., Xing Y., Wang Y., Owen D.L., Schenkel J.M., Boomer J.S., Green J.M., Yagita H., Chi H., Hogquist K.A, & Farrar M.A. (2014). Tumor necrosis factor receptor superfamily costimulation couples T cell receptor signal strength to thymic regulatory T cell differentiation. Nature immunology, 15(5), 473-481.

+ Open protocol

+ Expand

Transplantation of Genetically Modified Bone Marrow

Check if the same lab product or an alternative is used in the 5 most similar protocols

Rag2^−/− recipient mice were sublethally irradiated 24 hours prior to engraftment using an X-ray irradiator (Radsource RS2000 Biological Irradiator, Radsource, Suwanee, GA) and administering two doses of 250 rads separated by a two-hour rest. Bone marrow was harvested from tibias and femurs of donor mice, dissociated into single cell suspensions, and magnetically depleted of contaminating mature T and B cells by labeling with biotinylated anti-CD3, CD4, CD8, CD19, and CD45R antibodies (eBioscience) and secondarily with streptavidin-conjugated microbeads (Miltenyi Biotec) prior to passage through magnetic columns. Cell suspensions were washed in PBS and injected intravenously via the tail vein. Chimeric recipients were engrafted for 10-18 weeks prior to analysis.

+ Open protocol

+ Expand

CD8 T Cell Activation Assays

Check if the same lab product or an alternative is used in the 5 most similar protocols

1 × 10⁶ WT and KO CD8 T cells were stimulated in vitro with 2 μg/ml anti-CD28 (eBioscience) for the indicated amount of time in a 24-well dish pre-coated with 10 μg/ml anti-CD3 (eBioscience). Naïve cells received 10 ng/ml IL-7 for maintenance. Cells were negatively selected for CD8+ over columns (Miltenyi) prior to plating using biotinalyted antibodies and streptavidin conjugated microbeads (Miltenyi). For TopFluor-LPC assays (Avanti Polar Lipids), 0.1 μM TF-LPC was added during the last four hours of culture. Cell trace violet assays were performed according to the manufacturer’s protocol (eBioscience). All cell culture was performed in complete T cell media.

Piccirillo A.R., Hyzny E.J., Beppu L.Y., Menk A.V., Wallace C., Hawse W.F., Buechel H.M., Wong B.H., Chin Foo J., Cazenave-Gassiot A., Wenk M.R., Delgoffe G.M., Watkins S., Silver D.L, & D’Cruz L.M. (2019). The lysophosphatidylcholine transporter, MFSD2A, is essential for CD8+ memory T cell maintenance and secondary response to infection. Journal of immunology (Baltimore, Md. : 1950), 203(1), 117-126.

+ Open protocol

+ Expand

Isolation and Flow Cytometric Analysis of Murine Thymocytes

Check if the same lab product or an alternative is used in the 5 most similar protocols

Thymocytes were isolated by mechanical disruption. For analyzing DN compartment, total thymocytes were incubated with antibodies against lineage markers such as Ter119, CD11b, Ly6G/Gr1, IgM, CD19, CD4, CD8α, TCRβ, TCRγδ, NK1.1 and CD49b and lineage positive cells were removed by BioMag goat anti-rat IgG (QIAGEN) or streptavidin conjugated microbeads (Miltenyi Biotec). Total thymocytes or cells after depletion were labeled with fluorochrome-conjugated antibodies against T-, B-, NK-, ILC- cell markers for phenotypic analysis. Flow cytometric analysis was performed using a two-laser FACSCanto^™ (BD Biosciences) or a three-laser FACS Aria (BD Biosciences). Cell sorting was performed using a three-laser FACS Fortessa (BD Biosciences) or a three-laser SONY SH800 Cell sorter (SONY Biotechnology). The resulting files were uploaded to FlowJo v10 (BD Biosciences) for further analysis.

Hassan A.A., Khan I.U., Abdulmawjood A, & Lämmler C. (2001). Evaluation of PCR Methods for Rapid Identification and Differentiation of Streptococcus uberis and Streptococcus parauberis. Journal of Clinical Microbiology, 39(4), 1618-1621.

+ Open protocol

+ Expand

Isolation and Co-culture of Myeloid Cells and T Cells for Immunosuppression Analysis

Check if the same lab product or an alternative is used in the 5 most similar protocols

CD11b⁺Gr1⁺ myeloid cells and MDSCs were isolated from BM cells obtained by flushing femurs and tibias of tumor-free or MM-bearing mice, respectively, using magnetic cell separation technique. Briefly, BM cells were labelled with biotin-anti-Gr-1 antibody (BD) followed by Streptavidin-conjugated MicroBeads (Miltenyi Biotec) and selection on LS column. Purity of isolated cell population was confirmed by flow cytometry and was more than 98%. All Gr1⁺ cells expressed CD11b marker.
Splenocytes from OT-1 mice (1×10⁶/mL) were stimulated with 0.5 μg/mL specific (SIINFEKL) or control (RGPGRAFVTI) peptide in the presence or absence of CD11b⁺Gr1⁺ cells isolated from the BM of vehicle control or 17β-estradiol-treated MM-bearing mice. T cell proliferation was evaluated using [3H]thymidine incorporation.

Ozerova M, & Nefedova Y. (2019). Estrogen promotes multiple myeloma through enhancing the immunosuppressive activity of MDSC. Leukemia & lymphoma, 60(6), 1557-1562.

+ Open protocol

+ Expand

Quantifying Treg Suppression of T Cells

Check if the same lab product or an alternative is used in the 5 most similar protocols

CD4+ T cells were purified from spleens and lymph nodes of BDC and BDC-Idd9.905 mice by depleting B220+, CD8+ and CD11b+ cells using magnetic beads (Miltenyi Biotec GmbH). The purified CD4+ T cells were subsequently stained with biotinylated anti-CD25 mAb (clone 7D4, BD Bioscience), followed by column purification using streptavidin-conjugated microbeads (Miltenyi Biotec GmbH). The CD4+CD25− fraction was used as effector T cells. The CD4+CD25+ fraction was further purified by positive selection using an MS column (Miltenyi Biotec GmbH) and utilized as Treg cells. CD4+CD25− T cells (2.5 × 10⁴ cells/well) were cultured in triplicate with serial dilutions of CD4+CD25+ T cells in the presence of irradiated (3,200 rads) spleen cells (5 × 10⁴ cells/well) from NOD.scid mice and BDC2.5 mimotope p79 (0.1 μg/ml) in 96-well plates. T cell proliferation was determined as described above. Suppression of T cell proliferation was calculated using the following formula: ((T cell proliferation in the absence of Tregs - T cell proliferation in the presence of Tregs) / T cell proliferation in the absence of Tregs) × 100.

Berry G.J., Frielle C., Luu T., Salzberg A.C., Rainbow D.B., Wicker L.S, & Waldner H. (2015). Genome-wide transcriptional analyses of islet-specific CD4+ T cells identify Idd9 genes controlling diabetogenic T cell function. Journal of immunology (Baltimore, Md. : 1950), 194(6), 2654-2663.

+ Open protocol

+ Expand

About PubCompare

Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.

We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.

However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.

Ready to get started?

Revolutionizing how scientists
search and build protocols!