Dynabeads sheep anti rat igg

Manufactured by Thermo Fisher Scientific

Sourced in United States

Dynabeads sheep anti-Rat IgG are uniform, superparamagnetic beads coated with sheep antibodies specific for rat immunoglobulin G (IgG). They are designed for the isolation and purification of rat IgG from complex samples.

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Lab products found in correlation

Trizol, by Thermo Fisher Scientific (5 mentions) Lsrfortessa, by BD (4 mentions) Anti cd4 rat igg2b clone gk1, by BioXCell (3 mentions) Anti cd8a rat igg2a clone 53 6.72, by BioXCell (3 mentions) Dynabead, by Thermo Fisher Scientific (3 mentions) Cd11b, by BioLegend (3 mentions) Collagenase a, by Merck Group (3 mentions) Collagenase a, by Roche (2 mentions) Ter119, by BioLegend (2 mentions) Ter119, by BD (2 mentions) Cell lysis buffer, by Cell Signaling Technology (2 mentions) Mouse anti armenian and syrian hamster igg abs, by BD (2 mentions) Facsaria iiu, by BD (2 mentions) Anti cd4, by Thermo Fisher Scientific (2 mentions) Anti cd4 gk1, by BioLegend (2 mentions) Rat anti mouse cd31, by BD (2 mentions) Collagenase type 1, by Thermo Fisher Scientific (2 mentions) Anti pecam 1 antibody, by BD (2 mentions) Trypsin edta, by Merck Group (2 mentions) Ebm 2 basal medium, by Lonza (2 mentions)

Close spelling variants of this product

Dynabeads (2112 citations) Sheep anti-rat Dynabeads (7 citations) Anti-rat Dynabeads (3 citations)

71 protocols using dynabeads sheep anti rat igg

Isolation and Culture of Murine Endothelial and PDGFRβ+ Cells

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Tibiae and femurs from wild-type mice were collected in sterile Ca²⁺ and Mg²⁺ free PBS, crushed with mortar and pestle, subjected to collagenase digestion, filtered and washed thrice to obtain a single cell suspension. Endothelial cells were then sorted using Endomucin antibody (cat. no. SC-65495) and Dynabeads sheep anti-Rat IgG (Invitrogen). Sorted ECs were then plated on dishes coated with fibronectin and cultured in endothelial cell growth medium (EBM-2, Clonetics; Lonza) supplemented with EGM-2 SingleQuots (CC-4176, Clonetic; Lonza).
PDGFRβ+ cells were sorted from single cell suspensions using CD140b/PDGFB Receptor β antibody (eBioscience, cat. no. 14-1402-82) and Dynabeads sheep anti-Rat IgG (Invitrogen). Sorted PDGFRβ+ cells were cultured on tissue culture plates containing in alpha MEM (Gibco) and 10% fetal bovine serum (Gibco).
Cultures of ECs or PDGFRβ+ cells were maintained at 37°C with 5% CO2 in a humidified atmosphere. For DFM treatment and subsequent analysis, cultures between passage 1 and 2 were used. Cells were treated with DFM (6.25 mg/ml of culture medium) for the duration of 36 hrs and subsequently cell culture medium (directly) or cells after trypsinization, three washings and lysate preparation were used for Enzyme-Linked Immunosorbent Assay.

Kusumbe A.P., Ramasamy S.K., Itkin T., Andaloussi Mäe M., Langen U.H., Betsholtz C., Lapidot T, & Adams R.H. (2016). Age-dependent modulation of vascular niches for haematopoietic stem cells. Nature, 532(7599), 380-384.

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Isolation of Lung and Liver Endothelial Cells

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Lungs and liver from Eng^flox/flox pups were surgically removed, rinsed in ice-cold DMEM and endothelial cells were isolated as described [46 (link)]. Briefly, tissues were minced by the use of scissors, digested in DMEM-3 mg.ml⁻¹ Collagenase A (10103586001, Roche, Basel, Switzerland) for 15 min at 37 °C and then filtered through a 70-µm strainer. The cell suspension was centrifuged at 200× g for 5 min and CD45⁺ cells were removed using Dynabeads sheep anti-Rat IgG (11035, Invitrogen, Waltham, MA, USA) coated with rat anti-mouse CD45 antibody (550539, BD Pharmigen, Franklin Lake, NJ, USA). Endothelial cells were sorted using Dynabeads sheep anti-Rat IgG (11035, Invitrogen, Waltham, MA, USA) coated with rat anti-mouse PECAM1 antibody (550274, BD Pharmigen, Franklin Lake, NJ, USA) according to the manufacturer’s instructions. After washing 5 times with DMEM-0.1% BSA, cells were seeded in 6-well plates. Lung and liver ECs were maintained for two-three passages in Endothelial Cell Growth Medium 2 (C-22011, PromoCell, Heidelberg, Germany) complemented with Fetal Calf Serum, Human Epidermal Growth Factors, Basic Fibroblast Growth Factor, Insulin Like Growth Factor, Human Vascular Endothelial Growth Factor-165, Ascorbic acid, Heparin and Hydrocortisone (C-39211, SupplementPack EC GM2, PromoCell, Heidelberg, Germany).

Galaris G., Montagne K., Thalgott J.H., Goujon G.J., van den Driesche S., Martin S., Mager H.J., Mummery C.L., Rabelink T.J, & Lebrin F. (2021). Thresholds of Endoglin Expression in Endothelial Cells Explains Vascular Etiology in Hereditary Hemorrhagic Telangiectasia Type 1. International Journal of Molecular Sciences, 22(16), 8948.

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Mature Megakaryocyte Generation Protocol

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Bone marrow-derived mature megakaryocytes were generated as described [25 (link)]. Briefly, mouse femurs were flushed, and cells expressing Ly6G/Ly6C, CD11b, CD16/32, and B220 were depleted using magnetic beads (sheep anti-rat IgG Dynabeads, Thermo Fisher Scientific, Waltham, MA, USA) and the following antibodies: anti-mouse Ly6G/Ly6C (561103, BD Biosciences, Franklin Lakes, NJ, USA), anti-mouse CD11b (47-0112-82, Thermo Fisher Scientific), anti-mouse CD16/CD32 (553141, BD Biosciences), and anti-mouse B220 (BD Biosciences). Negatively selected cells were incubated in megakaryocyte medium (Stempro-34 SFM, Thermo Fisher Scientific) containing 2.6% nutrient supplement, 1% glutamine, 1% penicillin–streptomycin–fungizone, and 20 ng/mL stem cell factor (Peprotech EC Ltd., London, UK) for 2 d at 37 °C and 5% CO₂, followed by a 5-d incubation with additional 50 ng/mL thrombopoietin (TPO) (Peprotech EC Ltd.). Mature megakaryocytes were enriched with a gradient of 3%/1.5% BSA (PAA Laboratories, Fisher Scientific, Hampton, NH, USA) under gravity for 45 min at RT. Cells in the lower 25% of the gradient, representing mature megakaryocytes, were washed in PBS and harvested in TriFast™ (VWR, Radnor, PA, USA).

Goeritzer M., Schlager S., Kuentzel K.B., Vujić N., Korbelius M., Rainer S., Kolb D., Mussbacher M., Salzmann M., Schrottmaier W.C., Assinger A., Schlagenhauf A., Madreiter-Sokolowski C.T., Blass S., Eichmann T.O., Graier W.F, & Kratky D. (2022). Adipose Triglyceride Lipase Deficiency Attenuates In Vitro Thrombus Formation without Affecting Platelet Activation and Bleeding In Vivo. Cells, 11(5), 850.

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Isolation of Double-Negative T Cell Progenitors

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Fresh thymocytes were resuspended in isolation buffer (PBS without cations, pH 7.2–7.4, containing 0.1% BSA and 2 mM EDTA) on ice. For CD4+ CD8+ T cell depletion, 5 × 10⁷ thymocytes were incubated with 20 µg anti-CD4 (rat IgG_2b clone GK1.5; BioXCell, West Lebanon, NH) and 37.5 µg anti-CD8a (rat IgG_2a clone 53–6.72; BioXCell) in 5 ml isolation buffer for 20 min at 4°C with tilted rotation, centrifuged, resuspended in 5 ml isolation buffer, and incubated twice with 250 µl sheep anti-rat IgG Dynabeads (Thermo Fisher Scientific, Waltham, MA) for 30 min at 4°C with tilted rotation. After each incubation, the tube was placed in a magnet for 2 min, unbound DN T cell progenitors were combined in a new tube, centrifuged and RNA was extracted from the cell pellet with 1 ml TRIZOL® (Ambion, Carlsbad, CA ) as described below.

Song Y., Kumar V., Wei H.X., Qiu J, & Stanley P. (2015). Lunatic, Manic and Radical Fringe Each Promote T and B Cell Development. Journal of immunology (Baltimore, Md. : 1950), 196(1), 232-243.

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Isolation and Analysis of Murine Hematopoietic Cells

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BM, spleen and thymus were dissected, crushed in PBS with 2% FCS and passed through a 70µm filter. Cells were counted using a Sysmex hematology analyzer (Sysmex). Cells were incubated with Fc-blocking antibodies (CD16/32, clone: 93, BD) prior to staining with fluorescently labeled antibodies. For antibody panels see Table S1. Dead cell discrimination was performed using propidium iodide (PI), 7AAD or live/dead fixable Aqua dead cell stain (Thermo Fisher Scientific). For FACS sorting of progenitors from BM, mature cells were depleted using purified antibodies against TER119, CD19, CD3, GR1 and MAC1 in combination with sheep anti-rat IgG Dynabeads (Thermo Fisher Scientific) prior to the specific antibody staining. For FACS sorting of B cell progenitors, total BM cells were either depleted as described above (omitting CD19) or enriched using anti-B220 beads (Miltenyi) prior to the specific antibody staining. Flow cytometry and cell sorting were performed using mainly the LSRFortessa and FACSARIAIIu/III platforms (BD Biosciences). Analysis of FACS data was done using FlowJo (BD Biosciences) and statistical analysis performed in R.

Peña-Pérez L., Kharazi S., Frengen N., Krstic A., Bouderlique T., Hauenstein J., He M., Somuncular E., Li Wang X., Dahlberg C., Gustafsson C., Johansson A.S., Walfridsson J., Kadri N., Woll P., Kierczak M., Qian H., Westerberg L., Luc S, & Månsson R. (2022). FOXO Dictates Initiation of B Cell Development and Myeloid Restriction in Common Lymphoid Progenitors. Frontiers in Immunology, 13, 880668.

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Isolation of Primary Pulmonary Microvascular Endothelial Cells

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Primary pulmonary microvascular endothelial cells (PMVECs) were isolated from lung tissues in DHCR24 flox/flox-Tie2Cre⁻ mice and DHCR24 flox/flox-Tie2Cre⁺ mice, as previously described (26 (link)). Hundred microliter of sheep anti-rat IgG Dynabeads (Thermo Fisher, USA) were mixed with 5 µg anti-mouse PECAM-1 (BD Pharmingen, USA). The beads and antibody were incubated overnight at 4°C. Fresh mouse lung tissue was isolated and washed with PBS, before being transferred to cryopreservation tubes and cut into pieces with sterile scissors. The lung tissue fragments were digested with collagenase for 45 minutes in a 37°C shaker. The cells were filtered through sterile 70-μm nylon mesh and washed twice with 0.1% bovine serum albumin (BSA; Solarbio, China).
To allow binding to cells, 30 µL of coated Dynabeads were added to cells and incubated for 25 minutes at room temperature. All bead-bound cells were resuspended in DMEM containing 20% FBS before plating and purified with Anti-ICAM-2 (BD Pharmingen) antibody-conjugated Dynabeads when the cell confluences reached 70% to 80% or above.

Li H., Yang Z., Liang W., Nie H., Guan Y., Yang N., Ji T., Liu Y., Huang Y., Zhang L., Yan J, & Zhang C. (2024). DHCR24 Insufficiency Promotes Vascular Endothelial Cell Senescence and Endothelial Dysfunction via Inhibition of Caveolin-1/ERK Signaling. The Journals of Gerontology Series A: Biological Sciences and Medical Sciences, 79(4), glae059.

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Isolation of Double Negative T Cells

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Fresh thymocytes were resuspended in isolation buffer (PBS lacking cations, pH 7.2-7.4, containing 0.1% BSA, 2 mM EDTA and 1 mg/ml glucose) on ice. For T cell depletion, ~3-5x10⁷ thymocytes were incubated with 20 μg anti-CD4 (rat IgG2b clone GK1.5, BioXCell) and 37.5 μg anti-CD8a (rat IgG2a clone 53-6.72; BioXCell) in 5 ml buffer for 20 min at 4°C with tilted rotation. After centrifugation, cells were resuspended in 5 ml buffer, and incubated with 250 μl sheep anti-rat IgG Dynabeads (Thermo Fisher Scientific) for 30 min at 4°C with tilted rotation. The tube was placed in a magnet for 2 min, unbound cells were centrifuged and resuspended in 250 μl Dynabeads for a second 30 min incubation at 4°C. After Dynabeads removal, unbound DN T cells were centrifuged, counted and RNA was extracted from the cell pellet with 1 ml TRIZOL (Ambion) as described below.

Tanwar A, & Stanley P. (2023). Synergistic regulation of Notch signaling by different O-glycans promotes hematopoiesis. Frontiers in Immunology, 14, 1097332.

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Thymocyte Isolation and T Cell Depletion

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Fresh thymocytes were resuspended in isolation buffer (PBS lacking cations, pH 7.2-7.4, containing 0.1% BSA, 2 mM EDTA and 1 mg/ml glucose) on ice. For T cell depletion, ~3-5x10 7 thymocytes were incubated with 20 µg anti-CD4 (rat IgG2b clone GK1.5, BioXCell) and 37.5 µg anti-CD8a (rat IgG2a clone 53-6.72; BioXCell) in 5 ml buffer for 20 min at 4°C with tilted rotation. After centrifugation, cells were resuspended in 5 ml buffer, and incubated with 250 µl sheep anti-rat IgG Dynabeads (Thermo Fisher Scientific) for 30 min at 4°C with tilted rotation. The tube was placed in a magnet for 2 min, unbound cells were centrifuged and resuspended in 250 µl Dynabeads for a second 30 min incubation at 4°C. After Dynabeads removal, unbound cells were centrifuged, counted and RNA was extracted from the cell pellet with 1 ml TRIZOL (Ambion) as described below.

Tanwar A., & Stanley P. (2022). Synergistic regulation of Notch signaling by different O-glycans promotes hematopoiesis.

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Megakaryocyte Isolation from Mouse Bone Marrow

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Megakaryocytes were prepared as previously described (Dumon et al., 2006 (link); Mazharian et al., 2011 (link)). In brief, bone marrow cells were obtained from mouse femurs and tibias by flushing, and cells expressing lineage-specific surface markers (CD16/CD32, Gr1, B220, or CD11b) were depleted using immunomagnetic beads (sheep anti-rat IgG Dynabeads, Invitrogen). The remaining population was cultured in 2.6% serum-supplemented StemPro medium with 2 mM L-glutamine, penicillin/streptomycin, and 20 ng/mL of murine stem cell factor at 37°C under 5% CO₂ for 2 days, and for a further 4 days in the presence of stem cell factor and 50 ng/mL thrombopoietin (37°C, 5% CO₂). Mature megakaryocytes were then enriched using a 1.5%/3% bovine serum albumin gradient under gravity (1 g) for 45 min at room temperature.

Vögtle T., Sharma S., Mori J., Nagy Z., Semeniak D., Scandola C., Geer M.J., Smith C.W., Lane J., Pollack S., Lassila R., Jouppila A., Barr A.J., Ogg D.J., Howard T.D., McMiken H.J., Warwicker J., Geh C., Rowlinson R., Abbott W.M., Eckly A., Schulze H., Wright G.J., Mazharian A., Fütterer K., Rajesh S., Douglas M.R, & Senis Y.A. (2019). Heparan sulfates are critical regulators of the inhibitory megakaryocyte-platelet receptor G6b-B. eLife, 8, e46840.

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Enrichment and Sorting of Human Dendritic Cells

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Human DCs were enriched from single cell BM suspensions by first labeling with Ab specific for human CD3, CD14, CD19, CD20 (all from Beckman Coulter), CD34 (BD BioSciences), and mouse CD45 (BD BioSciences) and Ter119 (BioLegend) followed by depletion of bound cells using sheep anti-rat IgG Dynabeads (Invitrogen) as previously published (30 (link)). Cells were then labeled with Live Dead^® aqua, anti-mouse CD45 PerCP Cy5.5, anti-human CD45 APC Cy7, CD3/CD14/CD19/CD20 Pacific blue, HLA DR PE Cy7, CD123 PE or PerCP Cy5.5, CD1c FITC, and CD141 APC and sorted using a Moflo Astrios (Beckman Coulter) (Figure S2 in Supplementary Material).

Minoda Y., Virshup I., Leal Rojas I., Haigh O., Wong Y., Miles J.J., Wells C.A, & Radford K.J. (2017). Human CD141+ Dendritic Cell and CD1c+ Dendritic Cell Undergo Concordant Early Genetic Programming after Activation in Humanized Mice In Vivo. Frontiers in Immunology, 8, 1419.

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