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Accucheck microbeads

Manufactured by Thermo Fisher Scientific
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AccuCheck microbeads are a type of laboratory equipment used for various applications. They are small, spherical particles that can be coated with specific ligands or antibodies to serve as a platform for various assays and experiments. The core function of AccuCheck microbeads is to provide a consistent and reliable means of capturing, isolating, or detecting target analytes in a sample.

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Lab products found in correlation

Magnetized ls column, by Miltenyi Biotec (1 mentions) Anti pe or anti apc magnetic microbeads, by Miltenyi Biotec (1 mentions) Cd11b percp cy5, by BD (1 mentions) Fc block, by Merck Group (1 mentions) Cd11c percp cy5, by BD (1 mentions) Hanks balanced salt solution (hbss), by Corning (1 mentions) Cd44 alexafluor700, by Thermo Fisher Scientific (1 mentions) Cd19 percp cy5, by Cytek Biosciences (1 mentions) Cd8 brilliant violet 785, by BioLegend (1 mentions) Cd3ε fitc, by BD (1 mentions) Ls column, by Miltenyi Biotec (1 mentions)

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Accucheck

4 protocols using accucheck microbeads

1

Murine Thymus and Spleen Immune Cell Analysis

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Mouse thymus or spleens were processed into a single cell suspension and stained with fluorochrome-labeled antibodies for 30 min on ice. Antibodies used are listed per manufacturer: CD3ε (145-2C11), CD8α (53-6.7) (Tonbo biosciences), CD4 (RM4-5), CD69 (H1.2F3), Thy1.2 (30-H12), CD45.1 (A20) (Biolegend), CD11b (M1/70), CD11c (HL3), CD19 (1D3), CD53 (OX-79) (BD Pharmigen), CD44 (IM7), F4/80 (BM8), TCRβ (H57-597) (eBioscience). For thymocytes, viable cells (as identified by FSC and SSC) were gated and then analyzed as described in the figures. For splenocytes, viable cells (as identified by FSC and SSC) were gated on and T cells (CD3ε+) were identified that were lineage (CD19, CD11b, CD11c, F4/80) negative and then analyzed as described in the figure captions. Cells were counted using AccuCheck microbeads (Invitrogen). Data was acquired on a LSRII (Becton Dickinson) and analyzed using FlowJo (Treestar).
Martinez R.J., Morris A.B., Neeld D.K, & Evavold B.D. (2016). Targeted loss of SHP1 in murine thymocytes dampens TCR signaling late in selection. European journal of immunology, 46(9), 2103-2110.
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2

Isolation and Adoptive Transfer of Naive CD4+ T Cells

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Splenocytes from naive mice were collected and processed into a single-cell suspension. CD4+ T cells were purified following manufacturer instructions using the CD4+ T-cell negative isolation kit (Miltneyi Biotec). Purified CD4+ T cells were analysed by flow cytometry for purity and counted by flow cytometry using AccuCheck microbeads (Invitrogen). Purified CD4s were injected intravenously into recipient mice and immunized 24 h later. Park rate at 24 h was measured in TCRα−/− and found to be ∼20% (Supplementary Fig. 3).
Martinez R.J., Andargachew R., Martinez H.A, & Evavold B.D. (2016). Low-affinity CD4+ T cells are major responders in the primary immune response. Nature Communications, 7, 13848.
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3

Tetramer-Based Enrichment and Analysis of Antigen-Specific T Cells

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Tetramers and monomers were provided by the National Institute of Allergy and Infectious Diseases Tetramer Core Facility at Emory University or were a generous gift of Marc Jenkins. Tetramer enrichment and staining was performed as previously described67 (link). Briefly, mouse peripheral lymphoid organs (spleen and inguinal, para-aortic, brachial, axillary, cervical and mesenteric lymph nodes) were processed into a single-cell suspension. Cells were then stained with the respective tetramer (phycoerythrin (PE)- and/or allophycocyanin (APC)-conjugated, 4 μg ml−1 final concentration) for 60 min at room temperature, washed, stained with 50 μl of anti-PE or anti-APC magnetic microbeads for 30 min on ice (Miltenyi Biotec, Germany), washed and enriched on a magnetized LS column (Miltenyi Biotec). The bound and flow-through samples were then sampled to determine population counts using AccuCheck microbeads (Invitrogen, Carlsbad, CA, USA) and stained for analysis by flow cytometry. Antibodies used are show in Supplementary Table 1. For intracellular staining, cells were treated with the Tonbo or eBioscience Fixation and Permiabilization kits as per the manufacturer protocol. Samples were collected on an LSR II (Becton Dickinson) and analysed using FlowJo (Treestar, Ashland, OR, USA).
Martinez R.J., Andargachew R., Martinez H.A, & Evavold B.D. (2016). Low-affinity CD4+ T cells are major responders in the primary immune response. Nature Communications, 7, 13848.
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4

Tetramer-based Enrichment of Antigen-specific T Cells

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Tetramer enrichment was performed as previously described (32 (link)). In brief, spleen and peripheral lymph nodes (inguinal, lumbar, mesenteric, cervical, axillary and brachial) were harvested and processed into a single cell suspension by passing through a 100μm strainer. Cells were washed in cold 1X HBSS (Cellgro) and resuspended with 4μg/mL of tetramer PE conjugated NFM:I-Ab and/or APC conjugated MOG:I-Ab (NIH tetramer core (33 (link), 34 (link))) in Fc block (heat killed mouse and rat serum, Sigma-Aldrich) at a volume 2× the pellet volume. After 1hr at room temperature, cells were wash in cold FACS wash (0.1% BSA, 0.05% NaN3, 1x PBS) and resuspend in 200μL FACS wash plus 50μL of anti-PE and/or anti-APC beads (Miltenyi Biotec) and incubated for 30 min. on ice. Cells were then washed and enriched on a LS column (Miltenyi Biotec). Unbound and column bound cell numbers were determined with AccuCheck microbeads (Invitrogen) alongside with cell surface marker characterization performed with flow cytometry (LSR II, BD) and FlowJo software (Treestar). Antibodies used included; CD3ε-FITC (145-2C11, BD Pharmingen), CD11b- PerCP -Cy5.5 (M1/70, BD Pharmingen), CD11c-PerCP-Cy5.5 (HL3, BD Pharmingen), CD19- PerCP -Cy5.5 (1D3, Tonbo Biosciences), CD4-Brilliant Violet 510 (RM4-5, BioLegend), CD8-Brilliant Violet 785 (53-6.7, BioLegend), CD44-Alexa Fluor 700 (IM7, eBioscience).
Blanchfield L., Sabatino JJ J.r., Lawrence L, & Evavold B.D. (2017). NFM cross-reactivity to MOG does not expand a critical threshold level of high affinity T cells necessary for onset of demyelinating disease. Journal of immunology (Baltimore, Md. : 1950), 199(8), 2680-2691.
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