Figure S1
Figure S1
COMPANY FOCUS
COMPANY FOCUS
MACS® Technology—
setting standards in cell separation MACS® Technology— setting standards in cell separation Benefit from the gold standard The gold standard in cell separation—a COMPANY FOCUS hallmark MACS® Technology has evolved Specific—cell separation by MACS Technology
Methods
Methods
Methods
Methods
Whole brain
H2O
15% EtOH
H2O
Methods
60 days
10#male#C57# 5 male C57
15 male
H2O
15% EtOH
15% EtOH
H2O
H2O
15% EtOH
15 male saline# 15 male
otherWestern day Every day
Every PFC PFC 60 days 60 days other day 10#male#C57# 5 male C57
qPCR blot
Astrocytes
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MACS MicroBeads for indirect Cell separation strategies magnetic labeling
ACSA2: proprietary astrocyte antibody
cellintype–specific MicroBeads, researchers have a short 15-min incubation step. Reliable—MACSmagnetically Cell Separation is highly labeling the desired target cell the option to use a monoclonal or reliable, polyclonal achieving consistently optimal Magnetic separation type. and purities. primary antibody that recognizes arecoveries cell type– Labeled and unlabeled cells are separated in a MACS Column placed specific epitope of choice for cell separation. Depletion—unwanted cells can be removed by within the magnetic field of a MACS Separator, a strong permanent Cell separation strategies magnetic labeling and subsequent separation The primary antibody can be unconjugated, magnet. The flow-through is Positive selection—target cells can be collected as the non-magnetic, biotinylated, or f luorochrome-conjugated. from the unlabeled target cells. negative fraction. The cells can be isolated through positive selection (fig. 1) by used for downstream applications. magnetically Miltenyi Biotec provides MicroBeads thatlabeling the desired target cell specifically recognize the antibodytype. of choice Both strategies can easily be comDepletion—unwanted cellsseparation can be removed by Elution of the labeled cell fraction magnetic and subsequent separation resulting in indirect magnetic labeling of labeling the bined for the isolation of cell subpopulations. The separation column is removed from the unlabeled target cells. desired cell type.Target cell labeled with MACS Separator MACS Column used for downstream applications.
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90%
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7PMt
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specifically For detailed information visit
Target cell MACS MicroBead
In addition to direct magnetic labeling using cell type–specific MicroBeads, researchers have the option to use a monoclonal or polyclonal primary antibody that recognizes a cell type– specific epitope of choice for cell separation. The primary antibody can be unconjugated,
recognize the antibody of choice resulting in indirect magnetic labeling of the desired cell type. 7PMt 4 MACS & more
In addition to direct magnetic labeling using cell type–specific MicroBeads, researchers have the option to use a monoclonal or polyclonal primary antibody that recognizes a cell type– specific epitope of choice for cell separation. The primary antibody can be unconjugated, biotinylated, or f luorochrome-conjugated. Miltenyi Biotec provides MicroBeads that specifically recognize the antibody of choice www.miltenyibiotec.com/neuroscience resulting in indirect magnetic labeling of the MACS Column desired cell type.
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H2O
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qPCR
PFCqPCR
4 MACS & more
3 replicates of pooled samples/ qPCR Treatment (Group 1, 2, 3)
90% for protein 10% for RNA
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Western blot For detailed information visit www.macscellseparation.com
www.miltenyibiotec.com/neuroscience
4 Fractions (TH, CD11b+, ACSA2+, CD/AC-) 4 MACS & more
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MACS MicroBeads for indirect magnetic labeling
cell type–specific MicroBeads, researchers have cell type–specific MicroBeads, researchers have biotinylated, or f luorochrome-conjugated. experiments. the option to use the a monoclonal polyclonal Miltenyi Biotec provides MicroBeads that optionor to use a monoclonal or polyclonal primary antibody that recognizes a cell type– specifically recognize the antibody of choice primary antibody specific epitope of choice for cell separation. that recognizes a cell type– resulting in indirect magnetic labeling of the The primary antibody can be unconjugated, Target cell labeled with MACS Separator MACS Column specific epitope of choice for cell separation. desired cell type. MACS MicroBead www.miltenyibiotec.com/neuroscience biotinylated, or f luorochrome-conjugated. Miltenyi Biotec The provides MicroBeadsantibody that primary can be unconjugated, For detailed information visit 7PMt www.miltenyibiotec.com/neuroscience MACS & more specifically recognize the antibody of choice Figure 1 The principle of MACS Cell Separation. Positive selection is shown as an example. For depletion biotinylated, orthef luorochrome-conjugated. resulting in indirect magnetic labeling of the undesired cells are magnetically labeled and retained within the column during separation. The target www.macscellseparation.com MACS Separator MACS Column cells flow through and are collected as non-labeled fraction. desired cell type. Miltenyi Biotec provides MicroBeads that
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qPCR
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60 daysqPCR
Every day
Microglia
Western blot
15 male
2 treatments 15 male X (SAL, LPS) Every day
Western blot
MACS Technology— setting standards in cell separation
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Pool each group blot by treatment (n=3/treatment)
qPCR
qPCR
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H2O H2O Control group CIE group
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MACS® Technology— setting standards in cell separation
MACS® Technology— setting standards in cell separation
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into since its introduction in 1989. Easy—the conjugates of antibodies and Isolate is based on cell labeling with highly specific allow for easy magnetic cell isolation any cell type from any species, fast and easy, monoclonal antibodies, which recognize cell (see fig. 1). magnetic particles, i.e., MACS MicroBeads, 250 cellcell separation allow with for more easy than magnetic isolationreagents. type–specific antigens on the cell surface. Fast—the separation procedure takes as little A 1).pivotal addition to the expanding These antibodies are coupled to nanosized as 30 minutes to complete and cells are ready (see fig. portfolio are procedure the products isolation of superparamagnetic particles. for immediate use in downstream applications, Fast—the separation takes for as little as 30 neural minutescells. to complete and cells are ready such as flow cytometry or cell culture. for immediate use in downstream applications, Safe—MACS MicroBeads are non-toxic and such as flow cytometry or cell culture. Magnetic labeling biodegradable; removal of MicroBeads is not Safe—MACS MicroBeads are non-toxic and Cells of interest are labeled with required. MACS® MicroBeads, i.e., antibodies biodegradable; removal of MicroBeads is not Gentle—the gentle separation process yields coupled to superparamagnetic particles, required. viable and functionally active cells. in a short 15-min incubation step. Gentle—the gentle separation process yields The gold standard in cell separation—a Benefit from theCell gold standard Reliable—MACS Separation is highly Easy—the conjugates of antibodies and viable and functionally active cells. Specific—cell MACS Technology hallmark MACS® Technology has evolved COMPANY FOCUS reliable, separation achieving byconsistently optimal magnetic particles, i.e., MACS MicroBeads, Benefit from theCellgold standard of antibodies and Magnetic separation Reliable—MACS Separation is highly Easy—the conjugates on celland labeling with highly specific allow for easy magnetic cell isolation into since its introduction in 1989. Isolate is based recoveries purities. Specific—cell MACS Technology i.e., MACS MicroBeads, reliable, separation achieving byconsistently optimal magnetic particles, Labeled and unlabeled cells are anyfor cellseparated type from any species, fast and easy, monoclonal antibodies, which recognize cell (see fig. 1). is based on cell labeling with highly specific allow easy magnetic cell isolation in a MACS Column placed recoveries and purities.
The gold standard in cell separation—a Benefit from the gold standard COMPANY FOCUS hallmark MACS® Technology has evolved Specific—cell separation by MACS Technology into since its introduction in 1989. Isolate is based on cell labeling with highly specific any cell type from any species, fast and easy, monoclonal antibodies, which recognize cell with more than 250 cell separation reagents. type–specific antigens on the cell surface. A pivotal addition to the expanding These antibodies are coupled to nanosized portfolio are the products for isolation of superparamagnetic particles. neural cells.
7PMt
www.miltenyibiotec.com/neuroscience
COMPANY FOCUS
COMPANY FOCUS
MACS® Technology—
setting standards in cell separation MACS® Technology— setting standards in cell separation Benefit from the gold standard The gold standard in cell separation—a COMPANY FOCUS hallmark MACS® Technology has evolved Specific—cell separation by MACS Technology
Methods
Methods
Methods
Methods
Whole brain
H2O
15% EtOH
H2O
Methods
60 days
10#male#C57# 5 male C57
15 male
H2O
15% EtOH
15% EtOH
H2O
H2O
15% EtOH
15 male saline# 15 male
otherWestern day Every day
Every PFC PFC 60 days 60 days other day 10#male#C57# 5 male C57
qPCR blot
Astrocytes
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MACS MicroBeads for indirect Cell separation strategies magnetic labeling
ACSA2: proprietary astrocyte antibody
cellintype–specific MicroBeads, researchers have a short 15-min incubation step. Reliable—MACSmagnetically Cell Separation is highly labeling the desired target cell the option to use a monoclonal or reliable, polyclonal achieving consistently optimal Magnetic separation type. and purities. primary antibody that recognizes arecoveries cell type– Labeled and unlabeled cells are separated in a MACS Column placed specific epitope of choice for cell separation. Depletion—unwanted cells can be removed by within the magnetic field of a MACS Separator, a strong permanent Cell separation strategies magnetic labeling and subsequent separation The primary antibody can be unconjugated, magnet. The flow-through is Positive selection—target cells can be collected as the non-magnetic, biotinylated, or f luorochrome-conjugated. from the unlabeled target cells. negative fraction. The cells can be isolated through positive selection (fig. 1) by used for downstream applications. magnetically Miltenyi Biotec provides MicroBeads thatlabeling the desired target cell specifically recognize the antibodytype. of choice Both strategies can easily be comDepletion—unwanted cellsseparation can be removed by Elution of the labeled cell fraction magnetic and subsequent separation resulting in indirect magnetic labeling of labeling the bined for the isolation of cell subpopulations. The separation column is removed from the unlabeled target cells. desired cell type.Target cell labeled with MACS Separator MACS Column used for downstream applications.
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90%
4 MACS & more
7PMt
4
specifically For detailed information visit
Target cell MACS MicroBead
In addition to direct magnetic labeling using cell type–specific MicroBeads, researchers have the option to use a monoclonal or polyclonal primary antibody that recognizes a cell type– specific epitope of choice for cell separation. The primary antibody can be unconjugated,
recognize the antibody of choice resulting in indirect magnetic labeling of the desired cell type. 7PMt 4 MACS & more
In addition to direct magnetic labeling using cell type–specific MicroBeads, researchers have the option to use a monoclonal or polyclonal primary antibody that recognizes a cell type– specific epitope of choice for cell separation. The primary antibody can be unconjugated, biotinylated, or f luorochrome-conjugated. Miltenyi Biotec provides MicroBeads that specifically recognize the antibody of choice www.miltenyibiotec.com/neuroscience resulting in indirect magnetic labeling of the MACS Column desired cell type.
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H2O
Every day
Every day
qPCR
PFCqPCR
4 MACS & more
3 replicates of pooled samples/ qPCR Treatment (Group 1, 2, 3)
90% for protein 10% for RNA
7PMt
Western blot For detailed information visit www.macscellseparation.com
www.miltenyibiotec.com/neuroscience
4 Fractions (TH, CD11b+, ACSA2+, CD/AC-) 4 MACS & more
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MACS MicroBeads for indirect magnetic labeling
cell type–specific MicroBeads, researchers have cell type–specific MicroBeads, researchers have biotinylated, or f luorochrome-conjugated. experiments. the option to use the a monoclonal polyclonal Miltenyi Biotec provides MicroBeads that optionor to use a monoclonal or polyclonal primary antibody that recognizes a cell type– specifically recognize the antibody of choice primary antibody specific epitope of choice for cell separation. that recognizes a cell type– resulting in indirect magnetic labeling of the The primary antibody can be unconjugated, Target cell labeled with MACS Separator MACS Column specific epitope of choice for cell separation. desired cell type. MACS MicroBead www.miltenyibiotec.com/neuroscience biotinylated, or f luorochrome-conjugated. Miltenyi Biotec The provides MicroBeadsantibody that primary can be unconjugated, For detailed information visit 7PMt www.miltenyibiotec.com/neuroscience MACS & more specifically recognize the antibody of choice Figure 1 The principle of MACS Cell Separation. Positive selection is shown as an example. For depletion biotinylated, orthef luorochrome-conjugated. resulting in indirect magnetic labeling of the undesired cells are magnetically labeled and retained within the column during separation. The target www.macscellseparation.com MACS Separator MACS Column cells flow through and are collected as non-labeled fraction. desired cell type. Miltenyi Biotec provides MicroBeads that
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Western blot
qPCR
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60 daysqPCR
Every day
Microglia
Western blot
15 male
2 treatments 15 male X (SAL, LPS) Every day
Western blot
MACS Technology— setting standards in cell separation
setting standards in cell separation CD11b magne+c bead sor+ng ACSA2 magnetic bead sorting
Pool each group blot by treatment (n=3/treatment)
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