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Millipore macsplex buffer (mbp
$Differential centrifugation of whole blood efficiently separates different cell and vesicle populations. (a) Schematic of the differential centrifugation protocol used to separate red (RBC) and white blood cells (WBC), platelets (PL), apoptotic bodies (AB), large (LEV) and small extracellular vesicles (SEV), as well as soluble protein (SP) and flow through (FT). (b) Representative transmission electron microscopy images of vesicles enriched in AB, LEV and SEV fractions. Corresponding scale bars are inset. (c) Representative Western blots of blood components for histone H3 (H3), CD42A, cleaved Caspase 9 (CASP9), BAX, CD9 and CD81. A CD9 blotted membrane with vesicular and SP components was additionally overexposed. Molecular weight is indicated beside each blot. (d) Nanoparticle tracking analysis data showing the concentration of particles/ml at sizes ranging from 0–1 μm in LEV, SEV and SP fractions. Inset is a quantification of the concentration of particles from 400–600 nm found in each fraction. (d) Median CD9/63/81 fluorescence for 35 capture antibody coated beads from the <t>MACSplex</t> vesicle surface protein flow cytometry assay in the different blood components. The platelet fraction was diluted 1:10 before analysis$
Macsplex Buffer (Mbp, supplied by Millipore, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/product/macsplex+buffer/macs+buffer/pmc08485184-68-8-30
Average 90 stars, based on 1 article reviews

macsplex buffer (mbp - by Bioz Stars, 2026-10

90/100 stars

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1) Product Images from "Extracellular vesicles are the primary source of blood‐borne tumour‐derived mutant KRAS DNA early in pancreatic cancer"

Article Title: Extracellular vesicles are the primary source of blood‐borne tumour‐derived mutant KRAS DNA early in pancreatic cancer

Journal: Journal of Extracellular Vesicles

doi: 10.1002/jev2.12142

$Differential centrifugation of whole blood efficiently separates different cell and vesicle populations. (a) Schematic of the differential centrifugation protocol used to separate red (RBC) and white blood cells (WBC), platelets (PL), apoptotic bodies (AB), large (LEV) and small extracellular vesicles (SEV), as well as soluble protein (SP) and flow through (FT). (b) Representative transmission electron microscopy images of vesicles enriched in AB, LEV and SEV fractions. Corresponding scale bars are inset. (c) Representative Western blots of blood components for histone H3 (H3), CD42A, cleaved Caspase 9 (CASP9), BAX, CD9 and CD81. A CD9 blotted membrane with vesicular and SP components was additionally overexposed. Molecular weight is indicated beside each blot. (d) Nanoparticle tracking analysis data showing the concentration of particles/ml at sizes ranging from 0–1 μm in LEV, SEV and SP fractions. Inset is a quantification of the concentration of particles from 400–600 nm found in each fraction. (d) Median CD9/63/81 fluorescence for 35 capture antibody coated beads from the MACSplex vesicle surface protein flow cytometry assay in the different blood components. The platelet fraction was diluted 1:10 before analysis$
Figure Legend Snippet: Differential centrifugation of whole blood efficiently separates different cell and vesicle populations. (a) Schematic of the differential centrifugation protocol used to separate red (RBC) and white blood cells (WBC), platelets (PL), apoptotic bodies (AB), large (LEV) and small extracellular vesicles (SEV), as well as soluble protein (SP) and flow through (FT). (b) Representative transmission electron microscopy images of vesicles enriched in AB, LEV and SEV fractions. Corresponding scale bars are inset. (c) Representative Western blots of blood components for histone H3 (H3), CD42A, cleaved Caspase 9 (CASP9), BAX, CD9 and CD81. A CD9 blotted membrane with vesicular and SP components was additionally overexposed. Molecular weight is indicated beside each blot. (d) Nanoparticle tracking analysis data showing the concentration of particles/ml at sizes ranging from 0–1 μm in LEV, SEV and SP fractions. Inset is a quantification of the concentration of particles from 400–600 nm found in each fraction. (d) Median CD9/63/81 fluorescence for 35 capture antibody coated beads from the MACSplex vesicle surface protein flow cytometry assay in the different blood components. The platelet fraction was diluted 1:10 before analysis

Techniques Used: Centrifugation, Transmission Assay, Electron Microscopy, Western Blot, Molecular Weight, Concentration Assay, Fluorescence, Flow Cytometry

Magnetic Cell Separation:

Article Title: Stem cells implanted with nanofibrous mats for injured endometrial regeneration and immune-microenvironment remodeling
Article Snippet: .. Briefly, uteri were finely minced into ∼1-mm 3 pieces in a small volume of MACS buffer and pushed through a 60-mm mesh screen (Sigma-Aldrich, St. Louis, MO) using a syringe plunger. ..

Article Title: Vasculogenic mimicry structures in melanoma support the recruitment of monocytes
Article Snippet: Transfection efficiency was verified at 72 h via flow cytometry using ICAM-1-APC or IgG1-APC (both BD) in HUVE media. .. To isolate CD14+ monocytes, 107 MNCs were resuspended in 90 μl of MACS buffer (PBS, 0.5% BSA (SigmaAldrich), 2 mM EDTA (Sigma-Aldrich)) together with 10 μl of anti-CD14 microBeads (Miltenyi Biotec) for 15 min on ice. .. Cells were washed with MACS buffer and resuspended <2x108 cells/ml in MACS buffer prior to isolation via the autoMACS separator (Miltenyi Biotec) according to the “possels” program in the manufacturer’s instructions.

Article Title: Vasculogenic mimicry structures in melanoma support the recruitment of monocytes
Article Snippet: Transfection efficiency was verified at 72 h via flow cytometry using ICAM-1-APC or IgG1-APC (both BD) in HUVE media. .. To isolate CD14+ monocytes, 10 7 MNCs were resuspended in 90 μl of MACS buffer (PBS, 0.5% BSA (Sigma-Aldrich), 2 mM EDTA (Sigma-Aldrich)) together with 10 μl of anti-CD14 microBeads (Miltenyi Biotec) for 15 min on ice. .. Cells were washed with MACS buffer and resuspended <2x10 8 cells/ml in MACS buffer prior to isolation via the autoMACS separator (Miltenyi Biotec) according to the “possels” program in the manufacturer’s instructions.

Article Title: N-Acetylcysteine Alters Disease Progression and Increases Janus Kinase Mutation Frequency in a Mouse Model of Precursor B-Cell Acute Lymphoblastic Leukemia.
Article Snippet: .. The thymus was removed from euthanized mice, homogenized, and suspended in MACS buffer (500 mL 1x D-PBS; 1% 0.5M EDTA, pH 8.0; 2.5 g bovine serum albumin fraction V; MilliporeSigma). .. Single-cell suspensions were stained with phycoerythrinconjugated anti-CD19 antibody (clone 6D5; BioLegend, San Diego, CA) in preparation for flow cytometry.

Article Title: Co-evolution of human influenza A and Epstein Barr virus-specific CD8 ex vivo memory T cell receptor BV repertoires with increasing age
Article Snippet: .. The PBMC were counted and re-suspended in 20μl of anti-CD8 micro-beads (Miltenyi Biotech, Auburn, CA) and 80μl of MACS buffer [4°C Phosphate-buffered saline, 2.5g of Bovine Serum Albumin (Sigma-Aldrich, St.Louis, MO), 2ml 0.5M EDTA [pH 8.0] (Invitrogen, Grand Island, NY) degassed with sterile mesh filter] per 10 7 cells. ..

Article Title: Immune checkpoint status and exhaustion-related phenotypes of CD8 + T cells from the tumor-draining regional lymph nodes in breast cancer.
Article Snippet: The samples were run on a FACSAria II cell sorter (Becton Dickinson Biosciences) and analyzed by FlowJo software (Becton Dickinson Biosciences). .. The lymph node cells or PBMCs were stimulated with PMA (20 ng/mL) and ionomycin (0.5 μg/mL) (SigmaAldrich) in MACS buffer for 4 h at 37°C. .. Then, the cells were washed with ice- cold MACS buffer and labeled with IFN- γ and TNF- α catch reagents (Cytokine secretion assay kit, Miltenyi) for 10 min on ice.

Article Title: HX600, a synthetic agonist for RXR-Nurr1 heterodimer complex, prevents ischemia-induced neuronal damage
Article Snippet: The tissue was put back on ice, triturated and passed through 70 μm and 40 μm cell strainers (Falcon, Corning, NY, USA). .. Homogenates were centrifuged at 450 g for 5 min, resuspended in Miltenyi Myelin Debris Removal Beads II (Miltenyi, Cologne, Germany) and incubated at +4 °C for 15 min. After incubation the samples were washed with cold MACS buffer (PBS including 0.5% Bovine Serum Albumin, both from Sigma-Aldrich) and centrifuged at 400 g for 10 min, after which the pellets were resuspended into cold MACS buffer. .. Samples were applied to magnetic LD columns (Miltenyi) using 50 μm CellTrics pre-filters (Sysmex, Norderstedt, Germany), and the flow-through was collected on ice, counted and spun at 400 g for 10 min, and finally resuspended in RPMI-1640.

Article Title: Stem cells implanted with nanofibrous mats for injured endometrial regeneration and immune-microenvironment remodeling.
Article Snippet: .. Briefly, uteri were finely minced into ~1-mm3 pieces in a small volume of MACS buffer and pushed through a 60-mm mesh screen (Sigma-Aldrich, St. Louis, MO) using a syringe plunger. ..

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Journal: Journal of Extracellular Vesicles

Article Title: Extracellular vesicles are the primary source of blood‐borne tumour‐derived mutant KRAS DNA early in pancreatic cancer

doi: 10.1002/jev2.12142

Figure Lengend Snippet: Differential centrifugation of whole blood efficiently separates different cell and vesicle populations. (a) Schematic of the differential centrifugation protocol used to separate red (RBC) and white blood cells (WBC), platelets (PL), apoptotic bodies (AB), large (LEV) and small extracellular vesicles (SEV), as well as soluble protein (SP) and flow through (FT). (b) Representative transmission electron microscopy images of vesicles enriched in AB, LEV and SEV fractions. Corresponding scale bars are inset. (c) Representative Western blots of blood components for histone H3 (H3), CD42A, cleaved Caspase 9 (CASP9), BAX, CD9 and CD81. A CD9 blotted membrane with vesicular and SP components was additionally overexposed. Molecular weight is indicated beside each blot. (d) Nanoparticle tracking analysis data showing the concentration of particles/ml at sizes ranging from 0–1 μm in LEV, SEV and SP fractions. Inset is a quantification of the concentration of particles from 400–600 nm found in each fraction. (d) Median CD9/63/81 fluorescence for 35 capture antibody coated beads from the MACSplex vesicle surface protein flow cytometry assay in the different blood components. The platelet fraction was diluted 1:10 before analysis

Article Snippet: Beads were washed by adding 200 μl of MACSPlex Buffer (MBP) to each well and the filter plate was put on a vacuum manifold with vacuum applied at ‐100 mBar (Sigma‐Aldrich, Germany) until all wells were drained.

Techniques: Centrifugation, Transmission Assay, Electron Microscopy, Western Blot, Molecular Weight, Concentration Assay, Fluorescence, Flow Cytometry

Figure 1. Plasma and CSF preparation for MACSPlex human Exosome assay. (a) Protocol for extracellular vesicle (EV) enrichment and characterization by MACSPlex Exosome Assay. Blood and cerebrospinal ﬂuid (CSF) underwent serial centrifugation to eliminate cellular components and larger EVs. Samples were incubated overnight with phycoerythrin (PE)- and ﬂuorescein isothiocyanate (FITC)-labeled capture beads, coated with antibodies against 37 different EV surface markers. APC-conjugated detection antibodies against CD9, CD63, and CD81 were added and incubated for 1 h. After washing steps, samples were analyzed by ﬂow cytometry. (b) Schematic representation of the 37 EV surface markers analyzed by MACSPlex human Exosome assay.

Journal: Biomedicines

Article Title: Profiling Inflammatory Extracellular Vesicles in Plasma and Cerebrospinal Fluid: An Optimized Diagnostic Model for Parkinson's Disease.

doi: 10.3390/biomedicines9030230

Figure Lengend Snippet: Figure 1. Plasma and CSF preparation for MACSPlex human Exosome assay. (a) Protocol for extracellular vesicle (EV) enrichment and characterization by MACSPlex Exosome Assay. Blood and cerebrospinal ﬂuid (CSF) underwent serial centrifugation to eliminate cellular components and larger EVs. Samples were incubated overnight with phycoerythrin (PE)- and ﬂuorescein isothiocyanate (FITC)-labeled capture beads, coated with antibodies against 37 different EV surface markers. APC-conjugated detection antibodies against CD9, CD63, and CD81 were added and incubated for 1 h. After washing steps, samples were analyzed by ﬂow cytometry. (b) Schematic representation of the 37 EV surface markers analyzed by MACSPlex human Exosome assay.

Article Snippet: In total, 60 μL of plasma and 30 μL of ultracentrifuged CSF were added to the MACSPlex Buffer solution (final volume 120 μL) and analyzed with MACSQuant Analyzer-10 flow cytometer (Miltenyi, Bergisch Gladbach, Germany).

Techniques: Clinical Proteomics, Centrifugation, Incubation, Labeling, Cytometry

Figure 3. CSF-derived EVs characterization. Characterization of cerebrospinal ﬂuid (CSF)-derived extracellular vesicles (EVs) by nanoparticle tracking analysis (NTA) and MACSPlex human exosome assay ﬂow cytometry. Healthy controls (HC) were compared with patients with Parkinson’s disease (PD) multisystem atrophy (MSA), or atypical parkinsonism with tauopathies (AP-Tau). (a) EV concentration (n/mL CSF) at NTA. (b) EV diameter (nm) at NTA. Boxplots show median and interquartile range; bars show minimum and maximum values (* p < 0.05). (c) Normalized median ﬂuorescence intensity (nMFI; %) for 37 EV surface markers. Data and statistics are reported in Table S1. (d,e) Different perspectives of 3D-canonical plot reporting patient discrimination according to EV surface marker expression (each patient is indicated by a point and diagnoses are represented by colors: HC, blue; PD, red; MSA, orange; AP-Tau, grey). Canonical axes of the plot (canonical components 1, 2, and 3) are deﬁned by linear discrimination analysis from weighted linear combinations of the 37 EV markers analyzed by ﬂow cytometry. Spheres include patients with linear combination coefﬁcients that fall within the mean ± SD (canonicals 1, 2, and 3 ± SD).

Journal: Biomedicines

Article Title: Profiling Inflammatory Extracellular Vesicles in Plasma and Cerebrospinal Fluid: An Optimized Diagnostic Model for Parkinson's Disease.

doi: 10.3390/biomedicines9030230

Figure Lengend Snippet: Figure 3. CSF-derived EVs characterization. Characterization of cerebrospinal ﬂuid (CSF)-derived extracellular vesicles (EVs) by nanoparticle tracking analysis (NTA) and MACSPlex human exosome assay ﬂow cytometry. Healthy controls (HC) were compared with patients with Parkinson’s disease (PD) multisystem atrophy (MSA), or atypical parkinsonism with tauopathies (AP-Tau). (a) EV concentration (n/mL CSF) at NTA. (b) EV diameter (nm) at NTA. Boxplots show median and interquartile range; bars show minimum and maximum values (* p < 0.05). (c) Normalized median ﬂuorescence intensity (nMFI; %) for 37 EV surface markers. Data and statistics are reported in Table S1. (d,e) Different perspectives of 3D-canonical plot reporting patient discrimination according to EV surface marker expression (each patient is indicated by a point and diagnoses are represented by colors: HC, blue; PD, red; MSA, orange; AP-Tau, grey). Canonical axes of the plot (canonical components 1, 2, and 3) are deﬁned by linear discrimination analysis from weighted linear combinations of the 37 EV markers analyzed by ﬂow cytometry. Spheres include patients with linear combination coefﬁcients that fall within the mean ± SD (canonicals 1, 2, and 3 ± SD).

Techniques: Derivative Assay, Cytometry, Concentration Assay, Marker, Expressing

Figure 4. EV quantitative analysis: plasma vs. CSF. Quantitative analysis of extracellular vesicles (EVs) by nanoparticle tracking analysis (NTA) and MACSPlex assay ﬂow cytometry; plasma samples were compared to paired cerebrospinal ﬂuid (CSF) samples in patients with PD (Parkinson’s disease; n = 4), MSA (multisystem atrophy; n = 4) and AP-Tau (atypical parkinsonism with tauopathies; n = 4). (a) EV concentration (n/mL plasma or CSF) at NTA. (b) EV diameter (nm) at NTA. (c) MFI (expressed as arbitrary unity; a.u.) for CD9, CD63, CD81 and CD9-CD63-CD81 at ﬂow cytometry. Boxplots show median and interquartile range; bars show minimum and maximum values (** p < 0.05; *** p < 0.001). Data and statistics are reported in Table S2.

Journal: Biomedicines

Article Title: Profiling Inflammatory Extracellular Vesicles in Plasma and Cerebrospinal Fluid: An Optimized Diagnostic Model for Parkinson's Disease.

doi: 10.3390/biomedicines9030230

Figure Lengend Snippet: Figure 4. EV quantitative analysis: plasma vs. CSF. Quantitative analysis of extracellular vesicles (EVs) by nanoparticle tracking analysis (NTA) and MACSPlex assay ﬂow cytometry; plasma samples were compared to paired cerebrospinal ﬂuid (CSF) samples in patients with PD (Parkinson’s disease; n = 4), MSA (multisystem atrophy; n = 4) and AP-Tau (atypical parkinsonism with tauopathies; n = 4). (a) EV concentration (n/mL plasma or CSF) at NTA. (b) EV diameter (nm) at NTA. (c) MFI (expressed as arbitrary unity; a.u.) for CD9, CD63, CD81 and CD9-CD63-CD81 at ﬂow cytometry. Boxplots show median and interquartile range; bars show minimum and maximum values (** p < 0.05; *** p < 0.001). Data and statistics are reported in Table S2.

Techniques: Clinical Proteomics, Cytometry, Concentration Assay

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1) Product Images from "Extracellular vesicles are the primary source of blood‐borne tumour‐derived mutant KRAS DNA early in pancreatic cancer"

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