Rat anti mouse igm microbeads

Manufactured by Miltenyi Biotec

Sourced in United Kingdom

Rat anti-mouse IgM microbeads are a laboratory tool used for the isolation and enrichment of mouse IgM-positive cells. They consist of superparamagnetic beads coated with rat monoclonal antibodies specific for mouse IgM. These microbeads can be used in conjunction with magnetic separation techniques to efficiently isolate and purify IgM-expressing cells from complex biological samples.

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Lab products found in correlation

Penicillin streptomycin, by Lonza (1 mentions) Bovine serum albumin (bsa), by Merck Group (1 mentions) Foetal bovine serum (fbs), by Thermo Fisher Scientific (1 mentions) Human serum, by Merck Group (1 mentions) α mem, by Lonza (1 mentions) Lymphoprep, by Lonza (1 mentions) L ascorbic acid, by Thermo Fisher Scientific (1 mentions) Penicillin streptomycin, by Merck Group (1 mentions) Sodium pyruvate, by Merck Group (1 mentions) Cd14 microbead, by Miltenyi Biotec (1 mentions) Cd4 t cell isolation kit, by Miltenyi Biotec (1 mentions) Non essential amino acids (neaa), by Merck Group (1 mentions) L glutamine, by Merck Group (1 mentions) Human nk cell isolation kit, by Miltenyi Biotec (1 mentions) Automacs, by Miltenyi Biotec (1 mentions) Ficoll, by Harvard Bioscience (1 mentions) Pooled human ab serum, by Merck Group (1 mentions)

Spelling variants of this product

Anti-mouse IgM microbeads (14 citations) Anti-IgM MicroBeads (6 citations) Anti-rat microbeads, (3 citations)

4 protocols using rat anti mouse igm microbeads

Isolation and Culture of Bone Marrow SSCs

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A total of four patient samples were used for this study (samples from females
aged 51, 52, 56 and 71). Bone marrow from the four patients was utilised for
isolation and culture of bone marrow SSCs. SSC isolation has been described
previously.^{23 (link),24} Bone marrow was first washed and the solution was passed
through a 70-μm cell strainer and subjected to density centrifugation using
Lymphoprep™ (Lonza, Slough, UK). The buffy coat layer, containing bone marrow
mononuclear cells, was incubated with blocking buffer (α-MEM (Lonza), 10% human
serum (Sigma, Gillingham, UK), 5% foetal bovine serum (FBS) (Thermo Fisher
Scientific, Basingstoke, UK) and 10 mg/mL bovine serum albumin (BSA) (Sigma))
followed by washing with magnetic-activated cell sorting (MACS) buffer (PBS with
0.5% BSA and 2 mM ethylenediaminetetraacetic acid (EDTA; Sigma)). Cell
suspension was incubated with 1 mL STRO-1 antibody (generated in-house from
hybridoma). Following washing with MACS buffer, cells were incubated in rat
anti-mouse IgM microbeads (Miltenyi Biotec Ltd, Woking, UK). After further
washing with MACS buffer, target cell population was isolated by MACS and
resuspended in basal media (α-MEM containing 10% FBS and 1%
penicillin/streptomycin; Lonza) and plated into tissue culture flasks.

Waddell S.J., de Andrés M.C., Tsimbouri P.M., Alakpa E.V., Cusack M., Dalby M.J, & Oreffo R.O. (2018). Biomimetic oyster shell–replicated topography alters the behaviour of human skeletal stem cells. Journal of Tissue Engineering, 9, 2041731418794007.

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Isolation and Culture of Human Bone Marrow-Derived SSCs

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Bone marrow was obtained from patients undergoing total hip replacement surgery at Southampton General Hospital with full ethical consent and approval from the local hospital ethics committee (LREC 194/99/w, 27/10/10) and informed consent was obtained from all subjects. All methods utilising human tissue and cells were performed in accordance within the relevant guidelines and regulations. Bone marrow from 6 individual patients was collected and utilised for the isolation and culture of human bone marrow derived SSCs. Bone marrow was washed and the cell solution passed through a 70 μm cell filter strainer followed by treatment with Lymphoprep™ (Lonza). Isolated mononuclear cells were initialled incubated in blocking buffer (α-MEM, 10% human serum, 5% FCS and 10 mg/ml bovine serum albumin) and then washed with magnetic activated cell sorting (MACS) buffer (BSA and EDTA in PBS). Cells were then incubated in 1 ml of STRO-1 antibody (from hybridoma). Following washing with MACS buffer, cells were re-suspended in 1 ml containing 800 μl MACs buffer and 200 μl rat anti-mouse IgM microbeads (Miltenyi Biotec Ltd). Following washing with MACS buffer target cells were isolated by MACS. Following target cell isolation cells were washed and re-suspended in α-MEM containing 10% FCS and 1% penicillin/streptomycin (P/S) and placed into tissue culture flasks.

Budd E., de Andrés M.C., Sanchez-Elsner T, & Oreffo R.O. (2017). MiR-146b is down-regulated during the chondrogenic differentiation of human bone marrow derived skeletal stem cells and up-regulated in osteoarthritis. Scientific Reports, 7, 46704.

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Vitamin C Enhances CD34+ Cell Culture

Cited 1 time

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CD34+ cells were purified from baboon BM aspirates by immunomagnetic column purification using the anti-CD34 monoclonal antibody (clone 12.8; Dr. Ira Bernstein, Fred Hutchinson Cancer Research Center) and rat anti-mouse IgM microbeads (Miltenyi #130-047-302). For liquid cultures, cells were grown in Iscove's media containing 30% fetal bovine serum, 3 U/ml erythropoietin, 200 ng/ml stem cell factor, and 1 × 10⁻⁶ M dexamethasone. On days 1–3 culture media was also supplemented with IL-3 (10 ng/ml). For co-cultures, 2–3 × 10⁵ CD34+ cells were seeded onto monolayers of the ATF024 murine fetal liver stromal cell line (ATCC) and grown in the same media as for liquid cultures as previously described.^{44 (link)} In experiments designed to test the effect of Vitamin C, cells were cultured in media with and without Vitamin C (100 μM, L-ascorbic acid, Fisher #A61-25), a dose that showed maximum effect in dose-response experiments. A complete change of the media was performed every 2–3 days for both Vitamin C-supplemented and control cultures to maintain Vitamin C levels.

Ruiz M.A., Rivers A., Ibanez V., Vaitkus K., Mahmud N., DeSimone J, & Lavelle D. (2015). Hydroxymethylcytosine and demethylation of the γ-globin gene promoter during erythroid differentiation. Epigenetics, 10(5), 397-407.

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Isolation of Immune Cell Subsets

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Peripheral blood mononuclear cells (PBMCs) were isolated from buffy coats of healthy adult donors by density gradient purification with Ficoll (Biochrom). slanMo were isolated as described previously.^{3 (link)} Briefly, PBMCs were incubated in diluted M-DC8 hybridoma supernatant for 15 minutes at 4°C. After washing, the cells were incubated with rat anti-mouse IgM microbeads (Miltenyi) for 15 minutes at 4°C, washed again and then slanMo were purified by positive selection with an autoMACS (Miltenyi). Untouched NK cells from the remaining PBMCs were further purified using the human NK cell Isolation Kit (Miltenyi Biotech) according to the manufacturer’s instructions via autoMACS negative selection. The purity of slan⁺CD3⁻ slanMo and of CD56⁺CD3⁻ NK cells was ≥95%. Freshly isolated slanMo and NK cells were cultured in complete RPMI 1640 containing 2 mM L-glutamine, 1% penicillin/streptomycin, 1% non-essential amino acids (Sigma-Aldrich), 1 mM sodium pyruvate (Sigma-Aldrich), and 10% heat-inactivated pooled human AB serum (Sigma-Aldrich).
CD14 monocytes were isolated with CD14 microbeads (Miltenyi) by positive selection according to the protocol. T cell isolation was performed with the CD4 + T cell isolation kit (Miltenyi) by following the instructions and a negative selection autoMACS protocol.

Funck F., Pahl J., Kyjacova L., Freund L., Oehrl S., Gräbe G., Pezer S., Hassel J.C., Sleeman J., Cerwenka A, & Schäkel K. (2020). Human innate immune cell crosstalk induces melanoma cell senescence. Oncoimmunology, 9(1), 1808424.

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