concentrated protein Search Results


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  • 99
    Millipore mwco protein concentrators
    Mwco Protein Concentrators, supplied by Millipore, used in various techniques. Bioz Stars score: 99/100, based on 7 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    90
    Thermo Fisher pierce protein concentrator
    Pierce Protein Concentrator, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 90/100, based on 10 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Bio-Rad protein concerntration
    Protein Concerntration, supplied by Bio-Rad, used in various techniques. Bioz Stars score: 99/100, based on 22 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Thermo Fisher protein concentrator pes
    Protein Concentrator Pes, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 37 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Thermo Fisher protein concentrators
    Protein Concentrators, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 94/100, based on 4 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Millipore protein concentrator
    Protein Concentrator, supplied by Millipore, used in various techniques. Bioz Stars score: 85/100, based on 62 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Thermo Fisher pierce concentrators
    Pierce Concentrators, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 71 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Thermo Fisher 9k protein concentrators
    9k Protein Concentrators, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 19 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Thermo Fisher protein concentration total protein concentration
    PI3Kγ mediates innate immune memory cytokine response of microglia. Primary microglial cells (wild-type •, open columns; PI3Kγ −/− ■, dark gray columns; PI3Kγ KD/KD ▴, hatched columns) were primed with fixed priming doses of LPS [ULP (ultra-low dose, 1 fg/ml) and HP (high-dose, 100 ng/ml)] and restimulated on day 6 with a fixed dose LPS (100 ng/ml). Supernatants were collected 24 h after the second stimulation by LPS and the cytokine levels for (A) TNF-α, (B) IL-6, and (C) IL-10 were assayed by ELISA (normalized to <t>total</t> <t>protein</t> <t>concentration).</t> Data are shown as scatter dot plots, mean + SEM, n = 6, # p
    Protein Concentration Total Protein Concentration, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 4 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    93
    Millipore centricon protein concentrator
    PI3Kγ mediates innate immune memory cytokine response of microglia. Primary microglial cells (wild-type •, open columns; PI3Kγ −/− ■, dark gray columns; PI3Kγ KD/KD ▴, hatched columns) were primed with fixed priming doses of LPS [ULP (ultra-low dose, 1 fg/ml) and HP (high-dose, 100 ng/ml)] and restimulated on day 6 with a fixed dose LPS (100 ng/ml). Supernatants were collected 24 h after the second stimulation by LPS and the cytokine levels for (A) TNF-α, (B) IL-6, and (C) IL-10 were assayed by ELISA (normalized to <t>total</t> <t>protein</t> <t>concentration).</t> Data are shown as scatter dot plots, mean + SEM, n = 6, # p
    Centricon Protein Concentrator, supplied by Millipore, used in various techniques. Bioz Stars score: 93/100, based on 42 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Millipore centrifuge protein concentrator
    PI3Kγ mediates innate immune memory cytokine response of microglia. Primary microglial cells (wild-type •, open columns; PI3Kγ −/− ■, dark gray columns; PI3Kγ KD/KD ▴, hatched columns) were primed with fixed priming doses of LPS [ULP (ultra-low dose, 1 fg/ml) and HP (high-dose, 100 ng/ml)] and restimulated on day 6 with a fixed dose LPS (100 ng/ml). Supernatants were collected 24 h after the second stimulation by LPS and the cytokine levels for (A) TNF-α, (B) IL-6, and (C) IL-10 were assayed by ELISA (normalized to <t>total</t> <t>protein</t> <t>concentration).</t> Data are shown as scatter dot plots, mean + SEM, n = 6, # p
    Centrifuge Protein Concentrator, supplied by Millipore, used in various techniques. Bioz Stars score: 88/100, based on 21 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    86
    Sartorius AG vivaspin6 protein concentrators
    PI3Kγ mediates innate immune memory cytokine response of microglia. Primary microglial cells (wild-type •, open columns; PI3Kγ −/− ■, dark gray columns; PI3Kγ KD/KD ▴, hatched columns) were primed with fixed priming doses of LPS [ULP (ultra-low dose, 1 fg/ml) and HP (high-dose, 100 ng/ml)] and restimulated on day 6 with a fixed dose LPS (100 ng/ml). Supernatants were collected 24 h after the second stimulation by LPS and the cytokine levels for (A) TNF-α, (B) IL-6, and (C) IL-10 were assayed by ELISA (normalized to <t>total</t> <t>protein</t> <t>concentration).</t> Data are shown as scatter dot plots, mean + SEM, n = 6, # p
    Vivaspin6 Protein Concentrators, supplied by Sartorius AG, used in various techniques. Bioz Stars score: 86/100, based on 4 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Millipore amicon protein concentrator
    PI3Kγ mediates innate immune memory cytokine response of microglia. Primary microglial cells (wild-type •, open columns; PI3Kγ −/− ■, dark gray columns; PI3Kγ KD/KD ▴, hatched columns) were primed with fixed priming doses of LPS [ULP (ultra-low dose, 1 fg/ml) and HP (high-dose, 100 ng/ml)] and restimulated on day 6 with a fixed dose LPS (100 ng/ml). Supernatants were collected 24 h after the second stimulation by LPS and the cytokine levels for (A) TNF-α, (B) IL-6, and (C) IL-10 were assayed by ELISA (normalized to <t>total</t> <t>protein</t> <t>concentration).</t> Data are shown as scatter dot plots, mean + SEM, n = 6, # p
    Amicon Protein Concentrator, supplied by Millipore, used in various techniques. Bioz Stars score: 92/100, based on 55 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    84
    Millipore centriplus protein concentrators
    PI3Kγ mediates innate immune memory cytokine response of microglia. Primary microglial cells (wild-type •, open columns; PI3Kγ −/− ■, dark gray columns; PI3Kγ KD/KD ▴, hatched columns) were primed with fixed priming doses of LPS [ULP (ultra-low dose, 1 fg/ml) and HP (high-dose, 100 ng/ml)] and restimulated on day 6 with a fixed dose LPS (100 ng/ml). Supernatants were collected 24 h after the second stimulation by LPS and the cytokine levels for (A) TNF-α, (B) IL-6, and (C) IL-10 were assayed by ELISA (normalized to <t>total</t> <t>protein</t> <t>concentration).</t> Data are shown as scatter dot plots, mean + SEM, n = 6, # p
    Centriplus Protein Concentrators, supplied by Millipore, used in various techniques. Bioz Stars score: 84/100, based on 2 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Thermo Fisher icon protein concentrators
    PI3Kγ mediates innate immune memory cytokine response of microglia. Primary microglial cells (wild-type •, open columns; PI3Kγ −/− ■, dark gray columns; PI3Kγ KD/KD ▴, hatched columns) were primed with fixed priming doses of LPS [ULP (ultra-low dose, 1 fg/ml) and HP (high-dose, 100 ng/ml)] and restimulated on day 6 with a fixed dose LPS (100 ng/ml). Supernatants were collected 24 h after the second stimulation by LPS and the cytokine levels for (A) TNF-α, (B) IL-6, and (C) IL-10 were assayed by ELISA (normalized to <t>total</t> <t>protein</t> <t>concentration).</t> Data are shown as scatter dot plots, mean + SEM, n = 6, # p
    Icon Protein Concentrators, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 86/100, based on 14 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    GE Healthcare vivaspin protein concentrators
    PI3Kγ mediates innate immune memory cytokine response of microglia. Primary microglial cells (wild-type •, open columns; PI3Kγ −/− ■, dark gray columns; PI3Kγ KD/KD ▴, hatched columns) were primed with fixed priming doses of LPS [ULP (ultra-low dose, 1 fg/ml) and HP (high-dose, 100 ng/ml)] and restimulated on day 6 with a fixed dose LPS (100 ng/ml). Supernatants were collected 24 h after the second stimulation by LPS and the cytokine levels for (A) TNF-α, (B) IL-6, and (C) IL-10 were assayed by ELISA (normalized to <t>total</t> <t>protein</t> <t>concentration).</t> Data are shown as scatter dot plots, mean + SEM, n = 6, # p
    Vivaspin Protein Concentrators, supplied by GE Healthcare, used in various techniques. Bioz Stars score: 93/100, based on 19 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Thermo Fisher icontm protein concentrators
    PI3Kγ mediates innate immune memory cytokine response of microglia. Primary microglial cells (wild-type •, open columns; PI3Kγ −/− ■, dark gray columns; PI3Kγ KD/KD ▴, hatched columns) were primed with fixed priming doses of LPS [ULP (ultra-low dose, 1 fg/ml) and HP (high-dose, 100 ng/ml)] and restimulated on day 6 with a fixed dose LPS (100 ng/ml). Supernatants were collected 24 h after the second stimulation by LPS and the cytokine levels for (A) TNF-α, (B) IL-6, and (C) IL-10 were assayed by ELISA (normalized to <t>total</t> <t>protein</t> <t>concentration).</t> Data are shown as scatter dot plots, mean + SEM, n = 6, # p
    Icontm Protein Concentrators, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 86/100, based on 5 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    85
    Corning Life Sciences mwco protein concentrators
    PI3Kγ mediates innate immune memory cytokine response of microglia. Primary microglial cells (wild-type •, open columns; PI3Kγ −/− ■, dark gray columns; PI3Kγ KD/KD ▴, hatched columns) were primed with fixed priming doses of LPS [ULP (ultra-low dose, 1 fg/ml) and HP (high-dose, 100 ng/ml)] and restimulated on day 6 with a fixed dose LPS (100 ng/ml). Supernatants were collected 24 h after the second stimulation by LPS and the cytokine levels for (A) TNF-α, (B) IL-6, and (C) IL-10 were assayed by ELISA (normalized to <t>total</t> <t>protein</t> <t>concentration).</t> Data are shown as scatter dot plots, mean + SEM, n = 6, # p
    Mwco Protein Concentrators, supplied by Corning Life Sciences, used in various techniques. Bioz Stars score: 85/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Corning Life Sciences 10k protein concentrator
    PI3Kγ mediates innate immune memory cytokine response of microglia. Primary microglial cells (wild-type •, open columns; PI3Kγ −/− ■, dark gray columns; PI3Kγ KD/KD ▴, hatched columns) were primed with fixed priming doses of LPS [ULP (ultra-low dose, 1 fg/ml) and HP (high-dose, 100 ng/ml)] and restimulated on day 6 with a fixed dose LPS (100 ng/ml). Supernatants were collected 24 h after the second stimulation by LPS and the cytokine levels for (A) TNF-α, (B) IL-6, and (C) IL-10 were assayed by ELISA (normalized to <t>total</t> <t>protein</t> <t>concentration).</t> Data are shown as scatter dot plots, mean + SEM, n = 6, # p
    10k Protein Concentrator, supplied by Corning Life Sciences, used in various techniques. Bioz Stars score: 90/100, based on 2 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Sartorius AG vivaspin protein concentrator
    PI3Kγ mediates innate immune memory cytokine response of microglia. Primary microglial cells (wild-type •, open columns; PI3Kγ −/− ■, dark gray columns; PI3Kγ KD/KD ▴, hatched columns) were primed with fixed priming doses of LPS [ULP (ultra-low dose, 1 fg/ml) and HP (high-dose, 100 ng/ml)] and restimulated on day 6 with a fixed dose LPS (100 ng/ml). Supernatants were collected 24 h after the second stimulation by LPS and the cytokine levels for (A) TNF-α, (B) IL-6, and (C) IL-10 were assayed by ELISA (normalized to <t>total</t> <t>protein</t> <t>concentration).</t> Data are shown as scatter dot plots, mean + SEM, n = 6, # p
    Vivaspin Protein Concentrator, supplied by Sartorius AG, used in various techniques. Bioz Stars score: 89/100, based on 9 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    99
    Thermo Fisher pierce 20k mwco protein concentrators
    PI3Kγ mediates innate immune memory cytokine response of microglia. Primary microglial cells (wild-type •, open columns; PI3Kγ −/− ■, dark gray columns; PI3Kγ KD/KD ▴, hatched columns) were primed with fixed priming doses of LPS [ULP (ultra-low dose, 1 fg/ml) and HP (high-dose, 100 ng/ml)] and restimulated on day 6 with a fixed dose LPS (100 ng/ml). Supernatants were collected 24 h after the second stimulation by LPS and the cytokine levels for (A) TNF-α, (B) IL-6, and (C) IL-10 were assayed by ELISA (normalized to <t>total</t> <t>protein</t> <t>concentration).</t> Data are shown as scatter dot plots, mean + SEM, n = 6, # p
    Pierce 20k Mwco Protein Concentrators, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 16 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    95
    Fisher Scientific pierce protein concentrator
    PI3Kγ mediates innate immune memory cytokine response of microglia. Primary microglial cells (wild-type •, open columns; PI3Kγ −/− ■, dark gray columns; PI3Kγ KD/KD ▴, hatched columns) were primed with fixed priming doses of LPS [ULP (ultra-low dose, 1 fg/ml) and HP (high-dose, 100 ng/ml)] and restimulated on day 6 with a fixed dose LPS (100 ng/ml). Supernatants were collected 24 h after the second stimulation by LPS and the cytokine levels for (A) TNF-α, (B) IL-6, and (C) IL-10 were assayed by ELISA (normalized to <t>total</t> <t>protein</t> <t>concentration).</t> Data are shown as scatter dot plots, mean + SEM, n = 6, # p
    Pierce Protein Concentrator, supplied by Fisher Scientific, used in various techniques. Bioz Stars score: 95/100, based on 10 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    85
    Millipore ultrafree protein concentrator
    PI3Kγ mediates innate immune memory cytokine response of microglia. Primary microglial cells (wild-type •, open columns; PI3Kγ −/− ■, dark gray columns; PI3Kγ KD/KD ▴, hatched columns) were primed with fixed priming doses of LPS [ULP (ultra-low dose, 1 fg/ml) and HP (high-dose, 100 ng/ml)] and restimulated on day 6 with a fixed dose LPS (100 ng/ml). Supernatants were collected 24 h after the second stimulation by LPS and the cytokine levels for (A) TNF-α, (B) IL-6, and (C) IL-10 were assayed by ELISA (normalized to <t>total</t> <t>protein</t> <t>concentration).</t> Data are shown as scatter dot plots, mean + SEM, n = 6, # p
    Ultrafree Protein Concentrator, supplied by Millipore, used in various techniques. Bioz Stars score: 85/100, based on 3 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Bio-Rad protein concentrations
    PI3Kγ mediates innate immune memory cytokine response of microglia. Primary microglial cells (wild-type •, open columns; PI3Kγ −/− ■, dark gray columns; PI3Kγ KD/KD ▴, hatched columns) were primed with fixed priming doses of LPS [ULP (ultra-low dose, 1 fg/ml) and HP (high-dose, 100 ng/ml)] and restimulated on day 6 with a fixed dose LPS (100 ng/ml). Supernatants were collected 24 h after the second stimulation by LPS and the cytokine levels for (A) TNF-α, (B) IL-6, and (C) IL-10 were assayed by ELISA (normalized to <t>total</t> <t>protein</t> <t>concentration).</t> Data are shown as scatter dot plots, mean + SEM, n = 6, # p
    Protein Concentrations, supplied by Bio-Rad, used in various techniques. Bioz Stars score: 99/100, based on 4426 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Thermo Fisher mwco protein concentrator
    PI3Kγ mediates innate immune memory cytokine response of microglia. Primary microglial cells (wild-type •, open columns; PI3Kγ −/− ■, dark gray columns; PI3Kγ KD/KD ▴, hatched columns) were primed with fixed priming doses of LPS [ULP (ultra-low dose, 1 fg/ml) and HP (high-dose, 100 ng/ml)] and restimulated on day 6 with a fixed dose LPS (100 ng/ml). Supernatants were collected 24 h after the second stimulation by LPS and the cytokine levels for (A) TNF-α, (B) IL-6, and (C) IL-10 were assayed by ELISA (normalized to <t>total</t> <t>protein</t> <t>concentration).</t> Data are shown as scatter dot plots, mean + SEM, n = 6, # p
    Mwco Protein Concentrator, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 94/100, based on 12 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Millipore protein concentrate kit
    PI3Kγ mediates innate immune memory cytokine response of microglia. Primary microglial cells (wild-type •, open columns; PI3Kγ −/− ■, dark gray columns; PI3Kγ KD/KD ▴, hatched columns) were primed with fixed priming doses of LPS [ULP (ultra-low dose, 1 fg/ml) and HP (high-dose, 100 ng/ml)] and restimulated on day 6 with a fixed dose LPS (100 ng/ml). Supernatants were collected 24 h after the second stimulation by LPS and the cytokine levels for (A) TNF-α, (B) IL-6, and (C) IL-10 were assayed by ELISA (normalized to <t>total</t> <t>protein</t> <t>concentration).</t> Data are shown as scatter dot plots, mean + SEM, n = 6, # p
    Protein Concentrate Kit, supplied by Millipore, used in various techniques. Bioz Stars score: 99/100, based on 6 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Millipore concentrated protein solutions
    PI3Kγ mediates innate immune memory cytokine response of microglia. Primary microglial cells (wild-type •, open columns; PI3Kγ −/− ■, dark gray columns; PI3Kγ KD/KD ▴, hatched columns) were primed with fixed priming doses of LPS [ULP (ultra-low dose, 1 fg/ml) and HP (high-dose, 100 ng/ml)] and restimulated on day 6 with a fixed dose LPS (100 ng/ml). Supernatants were collected 24 h after the second stimulation by LPS and the cytokine levels for (A) TNF-α, (B) IL-6, and (C) IL-10 were assayed by ELISA (normalized to <t>total</t> <t>protein</t> <t>concentration).</t> Data are shown as scatter dot plots, mean + SEM, n = 6, # p
    Concentrated Protein Solutions, supplied by Millipore, used in various techniques. Bioz Stars score: 99/100, based on 2 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    87
    Sartorius AG vivaspin 20 protein concentrator
    PI3Kγ mediates innate immune memory cytokine response of microglia. Primary microglial cells (wild-type •, open columns; PI3Kγ −/− ■, dark gray columns; PI3Kγ KD/KD ▴, hatched columns) were primed with fixed priming doses of LPS [ULP (ultra-low dose, 1 fg/ml) and HP (high-dose, 100 ng/ml)] and restimulated on day 6 with a fixed dose LPS (100 ng/ml). Supernatants were collected 24 h after the second stimulation by LPS and the cytokine levels for (A) TNF-α, (B) IL-6, and (C) IL-10 were assayed by ELISA (normalized to <t>total</t> <t>protein</t> <t>concentration).</t> Data are shown as scatter dot plots, mean + SEM, n = 6, # p
    Vivaspin 20 Protein Concentrator, supplied by Sartorius AG, used in various techniques. Bioz Stars score: 87/100, based on 10 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    88
    Thermo Fisher mwco icon protein concentrator
    PI3Kγ mediates innate immune memory cytokine response of microglia. Primary microglial cells (wild-type •, open columns; PI3Kγ −/− ■, dark gray columns; PI3Kγ KD/KD ▴, hatched columns) were primed with fixed priming doses of LPS [ULP (ultra-low dose, 1 fg/ml) and HP (high-dose, 100 ng/ml)] and restimulated on day 6 with a fixed dose LPS (100 ng/ml). Supernatants were collected 24 h after the second stimulation by LPS and the cytokine levels for (A) TNF-α, (B) IL-6, and (C) IL-10 were assayed by ELISA (normalized to <t>total</t> <t>protein</t> <t>concentration).</t> Data are shown as scatter dot plots, mean + SEM, n = 6, # p
    Mwco Icon Protein Concentrator, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 88/100, based on 5 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Average 88 stars, based on 5 article reviews
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    mwco icon protein concentrator - by Bioz Stars, 2020-04
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    95
    Thermo Fisher 150 kd protein concentrator
    SMPD1 expression levels correlate with total SMase content in supernatant in MM cell lines and their exosomes. ( A ) Basal-dCT values of all four genes coding for SMases (SMPD1–4) measured by qRT-PCR in four human MM cell lines (JJN3, OPM2, LP1, and U266) after 24 h of culture. The housekeeping gene ABL1 was used for data normalization, and differential gene expression was determined using the comparative ΔΔCt method. ( B ) Basal total SMase content determined in supernatans by an AmplexRed SMase assay after 24 h of culture for all four cell lines. The amount of SMase activity measured in these cell lines corresponded best to SMPD1 expression, coding for acid SMase (ASM). ( C ) Cells and exosomes were isolated after 24 h of culture and analyzed for ASM content and exosomal markers, CD81 and CD63. Nanoparticle tracking analysis by Zetaview analysis shows a mean diameter of <t>50–150</t> nm, confirming the expected size of exosomes. Immunoblot can be found in Supplementary Figure S4B . ASM = acid sphingomyelinase, exo = exosomes. In A and B, the averages of n > 3 are shown.
    150 Kd Protein Concentrator, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 95/100, based on 15 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/150 kd protein concentrator/product/Thermo Fisher
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    PI3Kγ mediates innate immune memory cytokine response of microglia. Primary microglial cells (wild-type •, open columns; PI3Kγ −/− ■, dark gray columns; PI3Kγ KD/KD ▴, hatched columns) were primed with fixed priming doses of LPS [ULP (ultra-low dose, 1 fg/ml) and HP (high-dose, 100 ng/ml)] and restimulated on day 6 with a fixed dose LPS (100 ng/ml). Supernatants were collected 24 h after the second stimulation by LPS and the cytokine levels for (A) TNF-α, (B) IL-6, and (C) IL-10 were assayed by ELISA (normalized to total protein concentration). Data are shown as scatter dot plots, mean + SEM, n = 6, # p

    Journal: Frontiers in Immunology

    Article Title: Memory-Like Inflammatory Responses of Microglia to Rising Doses of LPS: Key Role of PI3Kγ

    doi: 10.3389/fimmu.2019.02492

    Figure Lengend Snippet: PI3Kγ mediates innate immune memory cytokine response of microglia. Primary microglial cells (wild-type •, open columns; PI3Kγ −/− ■, dark gray columns; PI3Kγ KD/KD ▴, hatched columns) were primed with fixed priming doses of LPS [ULP (ultra-low dose, 1 fg/ml) and HP (high-dose, 100 ng/ml)] and restimulated on day 6 with a fixed dose LPS (100 ng/ml). Supernatants were collected 24 h after the second stimulation by LPS and the cytokine levels for (A) TNF-α, (B) IL-6, and (C) IL-10 were assayed by ELISA (normalized to total protein concentration). Data are shown as scatter dot plots, mean + SEM, n = 6, # p

    Article Snippet: Measurement of the Protein Concentration Total protein concentration was determined using the Pierce™ 660 nm Protein Assay Kit (#22662) from Thermo Fisher Scientific (Massachusetts, USA).

    Techniques: Enzyme-linked Immunosorbent Assay, Protein Concentration

    Cytokine responses of microglia primed with increasing doses of LPS and β-glucan. (A) Schematic view of the two-step approach to induce memory-like inflammatory responses. Primary microglial cells (75,000 cells/well) isolated from neonatal mice were seeded on day 0 and stimulated with increasing doses of LPS (1 fg/ml to 100 ng/ml) or β-glucan (1 fg/ml to 1 μg/ml) on day 1 for 24 h. On day 2, cells were washed up and followed by medium change on day 4. Thereafter, cells were left to rest and restimulated on day 6 with a fixed dose of LPS (100 ng/ml). Six h and 24 h after the second stimulation, samples have been collected and processed for further analysis. (B–F) Opposing inflammatory response of microglia after priming with increasing doses of LPS or β-glucan. Microglial cells were stimulated with increasing doses of LPS ( B,C : 1 fg/ml to 100 ng/ml) or β-glucan ( D : 1 fg/ml to 1 μg/ml) for 24 h and rechallenged on day 6 with a fixed dose of LPS (100 ng/ml). Supernatants and protein lysates have been collected 24 h after the second stimulus by LPS (100 ng/ml). Cytokine levels ( B,D : TNF-α, n = 8; E : IL-6, n = 8; F : IL-10, n = 8) have been analyzed by ELISA (normalized to total protein concentration), whereas protein expression (C: TNF-α, n = 5) has been assayed by Western blotting and quantification (unprimed cells assigned as 1.0). Data are given as scatter dot plots, mean + SEM, # p

    Journal: Frontiers in Immunology

    Article Title: Memory-Like Inflammatory Responses of Microglia to Rising Doses of LPS: Key Role of PI3Kγ

    doi: 10.3389/fimmu.2019.02492

    Figure Lengend Snippet: Cytokine responses of microglia primed with increasing doses of LPS and β-glucan. (A) Schematic view of the two-step approach to induce memory-like inflammatory responses. Primary microglial cells (75,000 cells/well) isolated from neonatal mice were seeded on day 0 and stimulated with increasing doses of LPS (1 fg/ml to 100 ng/ml) or β-glucan (1 fg/ml to 1 μg/ml) on day 1 for 24 h. On day 2, cells were washed up and followed by medium change on day 4. Thereafter, cells were left to rest and restimulated on day 6 with a fixed dose of LPS (100 ng/ml). Six h and 24 h after the second stimulation, samples have been collected and processed for further analysis. (B–F) Opposing inflammatory response of microglia after priming with increasing doses of LPS or β-glucan. Microglial cells were stimulated with increasing doses of LPS ( B,C : 1 fg/ml to 100 ng/ml) or β-glucan ( D : 1 fg/ml to 1 μg/ml) for 24 h and rechallenged on day 6 with a fixed dose of LPS (100 ng/ml). Supernatants and protein lysates have been collected 24 h after the second stimulus by LPS (100 ng/ml). Cytokine levels ( B,D : TNF-α, n = 8; E : IL-6, n = 8; F : IL-10, n = 8) have been analyzed by ELISA (normalized to total protein concentration), whereas protein expression (C: TNF-α, n = 5) has been assayed by Western blotting and quantification (unprimed cells assigned as 1.0). Data are given as scatter dot plots, mean + SEM, # p

    Article Snippet: Measurement of the Protein Concentration Total protein concentration was determined using the Pierce™ 660 nm Protein Assay Kit (#22662) from Thermo Fisher Scientific (Massachusetts, USA).

    Techniques: Isolation, Mouse Assay, Enzyme-linked Immunosorbent Assay, Protein Concentration, Expressing, Western Blot

    SMPD1 expression levels correlate with total SMase content in supernatant in MM cell lines and their exosomes. ( A ) Basal-dCT values of all four genes coding for SMases (SMPD1–4) measured by qRT-PCR in four human MM cell lines (JJN3, OPM2, LP1, and U266) after 24 h of culture. The housekeeping gene ABL1 was used for data normalization, and differential gene expression was determined using the comparative ΔΔCt method. ( B ) Basal total SMase content determined in supernatans by an AmplexRed SMase assay after 24 h of culture for all four cell lines. The amount of SMase activity measured in these cell lines corresponded best to SMPD1 expression, coding for acid SMase (ASM). ( C ) Cells and exosomes were isolated after 24 h of culture and analyzed for ASM content and exosomal markers, CD81 and CD63. Nanoparticle tracking analysis by Zetaview analysis shows a mean diameter of 50–150 nm, confirming the expected size of exosomes. Immunoblot can be found in Supplementary Figure S4B . ASM = acid sphingomyelinase, exo = exosomes. In A and B, the averages of n > 3 are shown.

    Journal: Cancers

    Article Title: The Transfer of Sphingomyelinase Contributes to Drug Resistance in Multiple Myeloma

    doi: 10.3390/cancers11121823

    Figure Lengend Snippet: SMPD1 expression levels correlate with total SMase content in supernatant in MM cell lines and their exosomes. ( A ) Basal-dCT values of all four genes coding for SMases (SMPD1–4) measured by qRT-PCR in four human MM cell lines (JJN3, OPM2, LP1, and U266) after 24 h of culture. The housekeeping gene ABL1 was used for data normalization, and differential gene expression was determined using the comparative ΔΔCt method. ( B ) Basal total SMase content determined in supernatans by an AmplexRed SMase assay after 24 h of culture for all four cell lines. The amount of SMase activity measured in these cell lines corresponded best to SMPD1 expression, coding for acid SMase (ASM). ( C ) Cells and exosomes were isolated after 24 h of culture and analyzed for ASM content and exosomal markers, CD81 and CD63. Nanoparticle tracking analysis by Zetaview analysis shows a mean diameter of 50–150 nm, confirming the expected size of exosomes. Immunoblot can be found in Supplementary Figure S4B . ASM = acid sphingomyelinase, exo = exosomes. In A and B, the averages of n > 3 are shown.

    Article Snippet: The filtered medium was concentrated using a 150 kD Protein Concentrator (Thermo Scientific, Waltham, MA, USA) and filtered again with a 0.22 µM pore filter.

    Techniques: Expressing, Quantitative RT-PCR, Activity Assay, Isolation