rpmi1640  (Millipore)


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    Name:
    RPMI 1640 Vitamins Solution
    Description:
    Roswell Park Memorial Institute medium is commonly referred as RPMI It is a form of medium used in cell culture and tissue culture This medium contains phosphate and is formulated for use in a 5 carbon dioxide atmosphere RPMI 1640 is used for the serum free expansion of human lymphoid cells
    Catalog Number:
    r7256
    Price:
    None
    Applications:
    RPMI 1640 Vitamins Solution (100x) is used as a supplement in cell culture media.
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    Structured Review

    Millipore rpmi1640
    Concentration of sPLA 2 X released by PBMC of asthmatics and healthy controls. 2 × 10 6 PBMC were incubated in medium containing <t>RPMI1640</t> with 10% heat-inactivated FBS and antibiotics (37°C, 5% CO 2 ). Supernatants were collected after 24 hours of incubation in 37°C. The secretion of sPLA 2 X was measured in indirect ELISA. Data are presented as mean ± SEM; ∗ P
    Roswell Park Memorial Institute medium is commonly referred as RPMI It is a form of medium used in cell culture and tissue culture This medium contains phosphate and is formulated for use in a 5 carbon dioxide atmosphere RPMI 1640 is used for the serum free expansion of human lymphoid cells
    https://www.bioz.com/result/rpmi1640/product/Millipore
    Average 99 stars, based on 340 article reviews
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    rpmi1640 - by Bioz Stars, 2020-09
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    Images

    1) Product Images from "The Step Further to Understand the Role of Cytosolic Phospholipase A2 Alpha and Group X Secretory Phospholipase A2 in Allergic Inflammation: Pilot Study"

    Article Title: The Step Further to Understand the Role of Cytosolic Phospholipase A2 Alpha and Group X Secretory Phospholipase A2 in Allergic Inflammation: Pilot Study

    Journal: BioMed Research International

    doi: 10.1155/2014/670814

    Concentration of sPLA 2 X released by PBMC of asthmatics and healthy controls. 2 × 10 6 PBMC were incubated in medium containing RPMI1640 with 10% heat-inactivated FBS and antibiotics (37°C, 5% CO 2 ). Supernatants were collected after 24 hours of incubation in 37°C. The secretion of sPLA 2 X was measured in indirect ELISA. Data are presented as mean ± SEM; ∗ P
    Figure Legend Snippet: Concentration of sPLA 2 X released by PBMC of asthmatics and healthy controls. 2 × 10 6 PBMC were incubated in medium containing RPMI1640 with 10% heat-inactivated FBS and antibiotics (37°C, 5% CO 2 ). Supernatants were collected after 24 hours of incubation in 37°C. The secretion of sPLA 2 X was measured in indirect ELISA. Data are presented as mean ± SEM; ∗ P

    Techniques Used: Concentration Assay, Incubation, Indirect ELISA

    2) Product Images from "Analyses of Group III Secreted Phospholipase A2 Transgenic Mice Reveal Potential Participation of This Enzyme in Plasma Lipoprotein Modification, Macrophage Foam Cell Formation, and Atherosclerosis * Transgenic Mice Reveal Potential Participation of This Enzyme in Plasma Lipoprotein Modification, Macrophage Foam Cell Formation, and Atherosclerosis * S⃞"

    Article Title: Analyses of Group III Secreted Phospholipase A2 Transgenic Mice Reveal Potential Participation of This Enzyme in Plasma Lipoprotein Modification, Macrophage Foam Cell Formation, and Atherosclerosis * Transgenic Mice Reveal Potential Participation of This Enzyme in Plasma Lipoprotein Modification, Macrophage Foam Cell Formation, and Atherosclerosis * S⃞

    Journal: The Journal of Biological Chemistry

    doi: 10.1074/jbc.M804628200

    Foam cell formation from macrophages by PLA2G3, PLA2G5 and PLA2G10. A , LDL (2 mg/ml) was incubated with or without recombinant human PLA2G3 (sPLA 2 -III) and PLA2G5 (sPLA 2 -V) for 24 h at 37 °C. Then the NEFA released was quantified ( n = 2). B and C , J774 cells were cultured for 24 h in the presence or absence of LDL (200 μg/ml) that had been incubated for 24 h with or without PLA2G3 or PLA2G5. Then the cells were homogenized, and cholesteryl ester contents were quantified ( n = 2) ( B ). Replicate cells were fixed and stained with oil red O ( C ). Ac-LDL (50 μg/ml) was used as a positive control for foam cell formation. Magnification was ×200. D , peritoneal macrophages obtained from PLA2G3 Tg ( III-Tg ) mice and PLA2G10 Tg ( X-Tg ) mice as well as WT mice were placed in RPMI1640 containing 10% lipoprotein-deficient fetal bovine serum and were incubated with or without 200 μg/ml LDL or 50 μg/ml Ac-LDL. After 24 h, the cells were stained with oil red O (magnification, ×200). Magnified views of individual macrophages are shown in the inset. E , PGE 2 production by macrophages from PLA2G3 Tg ( left ) and PLA2G10 Tg ( right ) mice in comparison with WT mice; macrophages were incubated for 24 h in the presence (+) or absence (-) of 200 μg/ml LDL (mean ± S.E., n = 3–5; * , p
    Figure Legend Snippet: Foam cell formation from macrophages by PLA2G3, PLA2G5 and PLA2G10. A , LDL (2 mg/ml) was incubated with or without recombinant human PLA2G3 (sPLA 2 -III) and PLA2G5 (sPLA 2 -V) for 24 h at 37 °C. Then the NEFA released was quantified ( n = 2). B and C , J774 cells were cultured for 24 h in the presence or absence of LDL (200 μg/ml) that had been incubated for 24 h with or without PLA2G3 or PLA2G5. Then the cells were homogenized, and cholesteryl ester contents were quantified ( n = 2) ( B ). Replicate cells were fixed and stained with oil red O ( C ). Ac-LDL (50 μg/ml) was used as a positive control for foam cell formation. Magnification was ×200. D , peritoneal macrophages obtained from PLA2G3 Tg ( III-Tg ) mice and PLA2G10 Tg ( X-Tg ) mice as well as WT mice were placed in RPMI1640 containing 10% lipoprotein-deficient fetal bovine serum and were incubated with or without 200 μg/ml LDL or 50 μg/ml Ac-LDL. After 24 h, the cells were stained with oil red O (magnification, ×200). Magnified views of individual macrophages are shown in the inset. E , PGE 2 production by macrophages from PLA2G3 Tg ( left ) and PLA2G10 Tg ( right ) mice in comparison with WT mice; macrophages were incubated for 24 h in the presence (+) or absence (-) of 200 μg/ml LDL (mean ± S.E., n = 3–5; * , p

    Techniques Used: Incubation, Recombinant, Cell Culture, Staining, Positive Control, Mouse Assay

    3) Product Images from "Regulation of Bcl-2-family proteins in myeloma cells by three myeloma survival factors: interleukin-6, interferon-alpha and insulin-like growth factor 1"

    Article Title: Regulation of Bcl-2-family proteins in myeloma cells by three myeloma survival factors: interleukin-6, interferon-alpha and insulin-like growth factor 1

    Journal: Cell Death and Differentiation

    doi: 10.1038/sj.cdd.4400758

    Stat3, ERK1 and ERK2 phosphorylation in myeloma cells stimulated with IL-6, IFN-α or IGF-1 Myeloma cells were IL-6 and FCS starved for one hour and cultured with no cytokine (Co), 2 ng/mL IL-6 (IL-6), 200 U/mL IFN-α (IFN-α) or 100 ng/mL IGF-1 (IGF) in RPMI1640 and 1% BSA. Stat3 phosphorylation was analyzed by Western blot after 6 hours of culture using phospho-specific Stat3 antibody (P-Stat3), ERK1 and ERK2 phosphorylation after 15 minutes of culture using phospho-specific MAPK (P-MAPK). Blots were then stripped and reprobed for Stat3 or total MAPK. Western blots are for one representative experiment out of 3.
    Figure Legend Snippet: Stat3, ERK1 and ERK2 phosphorylation in myeloma cells stimulated with IL-6, IFN-α or IGF-1 Myeloma cells were IL-6 and FCS starved for one hour and cultured with no cytokine (Co), 2 ng/mL IL-6 (IL-6), 200 U/mL IFN-α (IFN-α) or 100 ng/mL IGF-1 (IGF) in RPMI1640 and 1% BSA. Stat3 phosphorylation was analyzed by Western blot after 6 hours of culture using phospho-specific Stat3 antibody (P-Stat3), ERK1 and ERK2 phosphorylation after 15 minutes of culture using phospho-specific MAPK (P-MAPK). Blots were then stripped and reprobed for Stat3 or total MAPK. Western blots are for one representative experiment out of 3.

    Techniques Used: Cell Culture, Western Blot

    Regulation of Bcl-2-family protein expression by IL-6, IFN-α or IGF-1 on XG-6 myeloma cells XG-6 myeloma cells were IL-6 and FCS starved for 1 hour and cultured for 2, 6 and 24 hours with no cytokine (Co), 2 ng/mL IL-6 (IL-6), 200 U/mL IFN-α (IFN-α) or 100 ng/mL IGF-1 (IGF) in RPMI1640 and 1% BSA. At the end of the culture, the cell count and the percentage of viable cells were assayed by trypan blue exclusion. The percentages of apoptotic cells were determined by staining with FITC-annexin V. Cells were immediately lysated and assayed for Bcl-2-family protein expression using Western blot analysis. The amount of proteins was carefully determined and 25 μg of total proteins from each culture group were loaded on the gel. In the experiments, MAPK expression was used as a loading protein control because it was not affected by the level of apoptosis. Western blots are for one representative experiment out of 3.
    Figure Legend Snippet: Regulation of Bcl-2-family protein expression by IL-6, IFN-α or IGF-1 on XG-6 myeloma cells XG-6 myeloma cells were IL-6 and FCS starved for 1 hour and cultured for 2, 6 and 24 hours with no cytokine (Co), 2 ng/mL IL-6 (IL-6), 200 U/mL IFN-α (IFN-α) or 100 ng/mL IGF-1 (IGF) in RPMI1640 and 1% BSA. At the end of the culture, the cell count and the percentage of viable cells were assayed by trypan blue exclusion. The percentages of apoptotic cells were determined by staining with FITC-annexin V. Cells were immediately lysated and assayed for Bcl-2-family protein expression using Western blot analysis. The amount of proteins was carefully determined and 25 μg of total proteins from each culture group were loaded on the gel. In the experiments, MAPK expression was used as a loading protein control because it was not affected by the level of apoptosis. Western blots are for one representative experiment out of 3.

    Techniques Used: Expressing, Cell Culture, Cell Counting, Staining, Western Blot

    IL-6, IFN-α and IGF-1 are myeloma cell survival factors XG-13 cells were harvested during the exponential growth phase with IL-6 and FCS, IL- 6 and FCS starved for 1 hour and then cultured for 2, 6 or 24 hours with no cytokine, 2 ng/mL of IL-6, 200 U/mL of IFN-α or 100 ng/mL of IGF-1 in RPMI1640 culture medium and 1% BSA. At the end of the culture, cells were stained with FITC-annexin V to determine the percentage of apoptotic cells with a FACScan.
    Figure Legend Snippet: IL-6, IFN-α and IGF-1 are myeloma cell survival factors XG-13 cells were harvested during the exponential growth phase with IL-6 and FCS, IL- 6 and FCS starved for 1 hour and then cultured for 2, 6 or 24 hours with no cytokine, 2 ng/mL of IL-6, 200 U/mL of IFN-α or 100 ng/mL of IGF-1 in RPMI1640 culture medium and 1% BSA. At the end of the culture, cells were stained with FITC-annexin V to determine the percentage of apoptotic cells with a FACScan.

    Techniques Used: Cell Culture, Staining

    4) Product Images from "HIV-1 competition experiments in humanized mice show that APOBEC3H imposes selective pressure and promotes virus adaptation"

    Article Title: HIV-1 competition experiments in humanized mice show that APOBEC3H imposes selective pressure and promotes virus adaptation

    Journal: PLoS Pathogens

    doi: 10.1371/journal.ppat.1006348

    Dynamics of hyper/hypo HIV-1 infection in intermediate A3H humanized mice. ( A, B ) HIV-1 infection in humanized mice. ( A ) A schematic of co-inoculation of hyper and hypo HIV-1s into intermediate A3H humanized mice. ( B ) Hyper and hypo viruses containing 2.5 ng of p24 antigen each (5 ng in total; n = 6) or RPMI1640 (n = 12; for mock infection) were inoculated into humanized mice. The amount of viral RNA in plasma (left) and the level of peripheral CD4 + T cells (CD45 + CD3 + CD4 + cells) (right) were analyzed at 0, 1, 2, 3, 5, and 6 wpi as described in Materials and Methods. The averages are shown in circles with SEMs, and the values from each mouse are shown by line. X-axes, wpi. In the left panel, horizontal broken line indicates detection limit (800 copies/ml plasma). In the right panel, * P
    Figure Legend Snippet: Dynamics of hyper/hypo HIV-1 infection in intermediate A3H humanized mice. ( A, B ) HIV-1 infection in humanized mice. ( A ) A schematic of co-inoculation of hyper and hypo HIV-1s into intermediate A3H humanized mice. ( B ) Hyper and hypo viruses containing 2.5 ng of p24 antigen each (5 ng in total; n = 6) or RPMI1640 (n = 12; for mock infection) were inoculated into humanized mice. The amount of viral RNA in plasma (left) and the level of peripheral CD4 + T cells (CD45 + CD3 + CD4 + cells) (right) were analyzed at 0, 1, 2, 3, 5, and 6 wpi as described in Materials and Methods. The averages are shown in circles with SEMs, and the values from each mouse are shown by line. X-axes, wpi. In the left panel, horizontal broken line indicates detection limit (800 copies/ml plasma). In the right panel, * P

    Techniques Used: Infection, Mouse Assay

    5) Product Images from "Vascular endothelial growth factor overproduced by tumour cells acts predominantly as a potent angiogenic factor contributing to malignant progression"

    Article Title: Vascular endothelial growth factor overproduced by tumour cells acts predominantly as a potent angiogenic factor contributing to malignant progression

    Journal: International Journal of Experimental Pathology

    doi: 10.1046/j.1365-2613.1999.00122.x

    (a) VEGF and (b) bFGF productivity of cancer cells transfected with VEGF cDNA. ▪ parental cells; □ transfected clones. The levels of VEGF and bFGF contained in conditioned medium and cell extract, respectively, were determined by sandwich ELISA. Sample values were plotted against a recombinant VEGF or bFGF standard curve (mean ± SE, n = 3). ND: not detectable. QG90, RPMI4788, MCF-7 and their transfected clones were cultured in RPMI1640 containing 10% FCS (normal). Hormones-stripped FCS with (E 2 +) or without (E 2 −) 10 −8 M 17 β-oestradiol was used for MCF-7 and its transfected clone, M2–24-G10.
    Figure Legend Snippet: (a) VEGF and (b) bFGF productivity of cancer cells transfected with VEGF cDNA. ▪ parental cells; □ transfected clones. The levels of VEGF and bFGF contained in conditioned medium and cell extract, respectively, were determined by sandwich ELISA. Sample values were plotted against a recombinant VEGF or bFGF standard curve (mean ± SE, n = 3). ND: not detectable. QG90, RPMI4788, MCF-7 and their transfected clones were cultured in RPMI1640 containing 10% FCS (normal). Hormones-stripped FCS with (E 2 +) or without (E 2 −) 10 −8 M 17 β-oestradiol was used for MCF-7 and its transfected clone, M2–24-G10.

    Techniques Used: Transfection, Clone Assay, Sandwich ELISA, Recombinant, Cell Culture

    6) Product Images from "High Density Lipoprotein Induces Proliferation and Migration of Human Prostate Androgen Independent Cancer Cells by an ABCA1-dependent Mechanism"

    Article Title: High Density Lipoprotein Induces Proliferation and Migration of Human Prostate Androgen Independent Cancer Cells by an ABCA1-dependent Mechanism

    Journal: Molecular cancer research : MCR

    doi: 10.1158/1541-7786.MCR-10-0008

    Activation of ERK1/2 in LNCaP by HDL after Overexpression of ABCA1 A. LNCaP cells were cultured in phenol red free RPMI1640 with 10 % FBS or charcoal stripped FBS for 72 h. Media was aspirated, and switched to phenol red free RPMI1640 with 1% FBS or charcoal stripped FBS for 24 h. Cells were then stimulated with HDL for 15 min. Cells were harvested and cell lysates were prepared for Western blotting. N; normal, CS; charcoal stripped. B. LNCaP cells were transfected with ABCA1 siRNA (A) or negative siRNA (N). After transfection, the cells were cultured in phenol red free RPMI1640 with 10 % charcoal stripped FBS for 72 h. Medium was aspirated, and switched to phenol red free RPMI1640 with 1% charcoal stripped FBS for 24 h. Cells were then stimulated with HDL (250 µg/mL) for 15 min, and harvested for Western blotting. Cells without transfection (M) and negative siRNA-transfected cells (N) were used as controls. The graph shows the ratio of pERK1/2 to ERK1/2 in each sample relative to 1.0 for lane 2 (n=3, means + s.d.).*P
    Figure Legend Snippet: Activation of ERK1/2 in LNCaP by HDL after Overexpression of ABCA1 A. LNCaP cells were cultured in phenol red free RPMI1640 with 10 % FBS or charcoal stripped FBS for 72 h. Media was aspirated, and switched to phenol red free RPMI1640 with 1% FBS or charcoal stripped FBS for 24 h. Cells were then stimulated with HDL for 15 min. Cells were harvested and cell lysates were prepared for Western blotting. N; normal, CS; charcoal stripped. B. LNCaP cells were transfected with ABCA1 siRNA (A) or negative siRNA (N). After transfection, the cells were cultured in phenol red free RPMI1640 with 10 % charcoal stripped FBS for 72 h. Medium was aspirated, and switched to phenol red free RPMI1640 with 1% charcoal stripped FBS for 24 h. Cells were then stimulated with HDL (250 µg/mL) for 15 min, and harvested for Western blotting. Cells without transfection (M) and negative siRNA-transfected cells (N) were used as controls. The graph shows the ratio of pERK1/2 to ERK1/2 in each sample relative to 1.0 for lane 2 (n=3, means + s.d.).*P

    Techniques Used: Activation Assay, Over Expression, Cell Culture, Western Blot, Transfection

    7) Product Images from "PDK1-mTOR signaling pathway inhibitors reduce cell proliferation in MK2206 resistant neuroblastoma cells"

    Article Title: PDK1-mTOR signaling pathway inhibitors reduce cell proliferation in MK2206 resistant neuroblastoma cells

    Journal: Cancer Cell International

    doi: 10.1186/s12935-015-0239-4

    Effect of AZD8055 (AZD), mTOR inhibitor, in MK-2206-resistant sublines compared with non-resistant cells. a Indicated cell lines were treated with AZD at indicated concentrations, with/without MK-2206 (5 μM) in RPMI1640 + 10 % FBS. Cell growth was evaluated as cell numbers at 72 h, and it was repeated three times. Data are expressed as the mean (±SD). b The effect of AZD on cell cycle phase distribution in LAN-1 and LAN-MK. Indicated cells were treated with AZD (50 nM) with/without MK-2206 (5 μM) in RPMI1640 with 10 % FBS for 12 h followed by analysis of cell cycle phase distribution, as introduced in methods and material. Cells were stained with PI for 30 min followed by FACScan flow cytometer. c IC 50 of AZD of indicated cell lines
    Figure Legend Snippet: Effect of AZD8055 (AZD), mTOR inhibitor, in MK-2206-resistant sublines compared with non-resistant cells. a Indicated cell lines were treated with AZD at indicated concentrations, with/without MK-2206 (5 μM) in RPMI1640 + 10 % FBS. Cell growth was evaluated as cell numbers at 72 h, and it was repeated three times. Data are expressed as the mean (±SD). b The effect of AZD on cell cycle phase distribution in LAN-1 and LAN-MK. Indicated cells were treated with AZD (50 nM) with/without MK-2206 (5 μM) in RPMI1640 with 10 % FBS for 12 h followed by analysis of cell cycle phase distribution, as introduced in methods and material. Cells were stained with PI for 30 min followed by FACScan flow cytometer. c IC 50 of AZD of indicated cell lines

    Techniques Used: Staining, Flow Cytometry, Cytometry

    Effect of AZD8055 (AZD) on mTOR-S6K axis in MK-2206-resistant sublines. a – d After 1 h serum starvation, indicated cell lines were incubated in RPMI1640 + 10 % FBS with/without MK-2206 (5 μM) or AZD (50 nM). Phosphorylation of mTOR and S6K were detected by western blot at 1.5 and 12 h, so was Actin. e Effect of U0126 on NB-19-MK cell line compared with NB-19. Indicated cells were treated with U0126 at indicated concentrations in RPMI1640 + 10 % FBS, with/without MK-2206 (5 μM). Cell growth was evaluated as cell number at 72 h, and result was repeated three times. Data are expressed as the mean (±SD). f NB-19 and NB-19-MK were incubated in RPMI1640 + 10 % FBS with/without MK-2206 (5 μM) or U0126 (5 μM). Phosphorylation of ERK, AKT, and S6K were detected by western blot at 1.5 and 12 h, so was Actin
    Figure Legend Snippet: Effect of AZD8055 (AZD) on mTOR-S6K axis in MK-2206-resistant sublines. a – d After 1 h serum starvation, indicated cell lines were incubated in RPMI1640 + 10 % FBS with/without MK-2206 (5 μM) or AZD (50 nM). Phosphorylation of mTOR and S6K were detected by western blot at 1.5 and 12 h, so was Actin. e Effect of U0126 on NB-19-MK cell line compared with NB-19. Indicated cells were treated with U0126 at indicated concentrations in RPMI1640 + 10 % FBS, with/without MK-2206 (5 μM). Cell growth was evaluated as cell number at 72 h, and result was repeated three times. Data are expressed as the mean (±SD). f NB-19 and NB-19-MK were incubated in RPMI1640 + 10 % FBS with/without MK-2206 (5 μM) or U0126 (5 μM). Phosphorylation of ERK, AKT, and S6K were detected by western blot at 1.5 and 12 h, so was Actin

    Techniques Used: Incubation, Western Blot

    MK-2206 suppressed the cell growth of NB cells. a MK-2206 suppressed the cell growth of NB cell lines. LAN-1, KP-N-SIFA, NB-19, and SK-N-DZ cells were cultured in RPMI1640 + 10 % FBS with MK-2206 at indicated concentrations. Cell growth was evaluated as cell numbers at 72 h, and it was repeated three times. Data are expressed as the mean (±SD). b Photomicrographs of MK-2206 non-resistant and resistant cells. Cells were cultured in glass bottom slide chambers with RPMI1640 + 10 % FBS, with MK-2206 (resistant sublines)/without MK-2206 (non-resistant cells) overnight. A 50 µm scale is indicated (Olympus Fluoview fv1000, DIC acquisition, ×40)
    Figure Legend Snippet: MK-2206 suppressed the cell growth of NB cells. a MK-2206 suppressed the cell growth of NB cell lines. LAN-1, KP-N-SIFA, NB-19, and SK-N-DZ cells were cultured in RPMI1640 + 10 % FBS with MK-2206 at indicated concentrations. Cell growth was evaluated as cell numbers at 72 h, and it was repeated three times. Data are expressed as the mean (±SD). b Photomicrographs of MK-2206 non-resistant and resistant cells. Cells were cultured in glass bottom slide chambers with RPMI1640 + 10 % FBS, with MK-2206 (resistant sublines)/without MK-2206 (non-resistant cells) overnight. A 50 µm scale is indicated (Olympus Fluoview fv1000, DIC acquisition, ×40)

    Techniques Used: Cell Culture

    MK-2206 showed less inhibition in cell growth of MK-2206-resistant sublines. a MK-2206 showed less inhibition in the proliferation of MK-2206-resistant sublines than in the non-resistant cells. Indicated cells were cultured in RPMI1640 + 10 % FBS with MK-2206 at indicated concentrations. Cell growth was evaluated as cell numbers at indicated hours, and it was repeated three times. Data are expressed as the mean (±SD). *P
    Figure Legend Snippet: MK-2206 showed less inhibition in cell growth of MK-2206-resistant sublines. a MK-2206 showed less inhibition in the proliferation of MK-2206-resistant sublines than in the non-resistant cells. Indicated cells were cultured in RPMI1640 + 10 % FBS with MK-2206 at indicated concentrations. Cell growth was evaluated as cell numbers at indicated hours, and it was repeated three times. Data are expressed as the mean (±SD). *P

    Techniques Used: Inhibition, Cell Culture

    Effect of GSK2334470 (GSK), PDK1 inhibitor, in MK-2206-resistant sublines compared with non-resistant cells. a Indicated cells were treated with GSK at indicated concentrations, with/without MK-2206 (5 μM) in RPMI1640 + 10 % FBS. Cell growth was evaluated as cell numbers at 72 h, and it was repeated three times. Data are expressed as the mean (±SD). b The effect of GSK on cell cycle phase distribution in LAN-1 and LAN-MK. LAN-1 and LAN-1-MK were treated with GSK (5 μM) with/without MK-2206 (5 μM) in RPMI1640 with 10 % FBS for 12 h followed by analysis of cell cycle phase distribution, as introduced in methods and material. Indicated cells were stained with PI for 30 min followed by FACScan flow cytometer. c IC 50 of GSK in indicated cells
    Figure Legend Snippet: Effect of GSK2334470 (GSK), PDK1 inhibitor, in MK-2206-resistant sublines compared with non-resistant cells. a Indicated cells were treated with GSK at indicated concentrations, with/without MK-2206 (5 μM) in RPMI1640 + 10 % FBS. Cell growth was evaluated as cell numbers at 72 h, and it was repeated three times. Data are expressed as the mean (±SD). b The effect of GSK on cell cycle phase distribution in LAN-1 and LAN-MK. LAN-1 and LAN-1-MK were treated with GSK (5 μM) with/without MK-2206 (5 μM) in RPMI1640 with 10 % FBS for 12 h followed by analysis of cell cycle phase distribution, as introduced in methods and material. Indicated cells were stained with PI for 30 min followed by FACScan flow cytometer. c IC 50 of GSK in indicated cells

    Techniques Used: Staining, Flow Cytometry, Cytometry

    Effect of GSK2334470 (GSK) on PDK1-mTOR-S6K axis in MK-2206-resistant sublines. a – d After 1 h serum starvation, indicated cells were incubated in RPMI1640 + 10 % FBS with/without MK-2206 (5 μM) or GSK (5 μM). Phosphorylation of PDK1, AKT, mTOR, and S6K were detected by western blot at 1.5 and 12 h, so were AKT and Actin. GSK3β, p-GSK3β and N-MYC were also detected
    Figure Legend Snippet: Effect of GSK2334470 (GSK) on PDK1-mTOR-S6K axis in MK-2206-resistant sublines. a – d After 1 h serum starvation, indicated cells were incubated in RPMI1640 + 10 % FBS with/without MK-2206 (5 μM) or GSK (5 μM). Phosphorylation of PDK1, AKT, mTOR, and S6K were detected by western blot at 1.5 and 12 h, so were AKT and Actin. GSK3β, p-GSK3β and N-MYC were also detected

    Techniques Used: Incubation, Western Blot

    8) Product Images from "Serum-Free Cryopreservation of Five Mammalian Cell Lines in Either a Pelleted or Suspended State"

    Article Title: Serum-Free Cryopreservation of Five Mammalian Cell Lines in Either a Pelleted or Suspended State

    Journal: Biological Procedures Online

    doi: 10.1251/bpo73

    Cells were grown to 50-100% confluency in a 25cm 2 flask. They were next trypsinized, and upon detachment, 3ml of growth medium were added. Cells were then split equally into three 15ml conical polycentrifuge tubes, followed by centrifugation and removal of trypsin solution. Each pellet was resuspended in 1 ml of the idicated cryopreservative (each with 10%DMSO: FBS=fetal bovine serum, PBS=phosphate-buffered saline, and RPMI = RPMI1640 with bicarbonate). One half of the cells from each treatment were removed to a new tube and placed in centrifuge to generate a pellet. Pelleted and suspended cells were then inserted into a foam box containing paper towels and placed in a -80°C freezer.
    Figure Legend Snippet: Cells were grown to 50-100% confluency in a 25cm 2 flask. They were next trypsinized, and upon detachment, 3ml of growth medium were added. Cells were then split equally into three 15ml conical polycentrifuge tubes, followed by centrifugation and removal of trypsin solution. Each pellet was resuspended in 1 ml of the idicated cryopreservative (each with 10%DMSO: FBS=fetal bovine serum, PBS=phosphate-buffered saline, and RPMI = RPMI1640 with bicarbonate). One half of the cells from each treatment were removed to a new tube and placed in centrifuge to generate a pellet. Pelleted and suspended cells were then inserted into a foam box containing paper towels and placed in a -80°C freezer.

    Techniques Used: Centrifugation

    9) Product Images from "Serum-Free Cryopreservation of Five Mammalian Cell Lines in Either a Pelleted or Suspended State"

    Article Title: Serum-Free Cryopreservation of Five Mammalian Cell Lines in Either a Pelleted or Suspended State

    Journal: Biological Procedures Online

    doi: 10.1251/bpo73

    Cells were grown to 50-100% confluency in a 25cm 2 flask. They were next trypsinized, and upon detachment, 3ml of growth medium were added. Cells were then split equally into three 15ml conical polycentrifuge tubes, followed by centrifugation and removal of trypsin solution. Each pellet was resuspended in 1 ml of the idicated cryopreservative (each with 10%DMSO: FBS=fetal bovine serum, PBS=phosphate-buffered saline, and RPMI = RPMI1640 with bicarbonate). One half of the cells from each treatment were removed to a new tube and placed in centrifuge to generate a pellet. Pelleted and suspended cells were then inserted into a foam box containing paper towels and placed in a -80°C freezer.
    Figure Legend Snippet: Cells were grown to 50-100% confluency in a 25cm 2 flask. They were next trypsinized, and upon detachment, 3ml of growth medium were added. Cells were then split equally into three 15ml conical polycentrifuge tubes, followed by centrifugation and removal of trypsin solution. Each pellet was resuspended in 1 ml of the idicated cryopreservative (each with 10%DMSO: FBS=fetal bovine serum, PBS=phosphate-buffered saline, and RPMI = RPMI1640 with bicarbonate). One half of the cells from each treatment were removed to a new tube and placed in centrifuge to generate a pellet. Pelleted and suspended cells were then inserted into a foam box containing paper towels and placed in a -80°C freezer.

    Techniques Used: Centrifugation

    10) Product Images from "FAM20B-catalyzed glycosaminoglycans control murine tooth number by restricting FGFR2b signaling"

    Article Title: FAM20B-catalyzed glycosaminoglycans control murine tooth number by restricting FGFR2b signaling

    Journal: BMC Biology

    doi: 10.1186/s12915-020-00813-4

    GAGs restrict the diffusion gradient of FGF10. a A hydrogel cell culture system was built to mimic the ECM environment. BaF3-FGFR2b cells were embedded in hydrogel with or without GAGs and cultured in RPMI1640 media. Cultures without GAGs in the hydrogel were divided into blank control or FGF10 control according to the presence or absence of 1000 pM FGF10 in the culture medium. The experimental groups were supplemented with 1000 pM FGF10 in the culture medium plus 1 μg/ml, 3 μg/ml, or 5 μg/ml of GAGs in the hydrogel. After 48 h of culture, the FGF activity of each group was calculated based on the viability of BaF3-FGFR2b cells (see Additional File 10: Fig. S10 ). b Cultures with GAGs in the hydrogel without FGF10 in the medium showed no significant differences from the blank control, indicating that GAGs alone did not affect the viability of BaF3 cells. c , d GAGs at 1 μg/ml and 3 μg/ml concentrations displayed differential config effects on FGF reactivity. e GAGs at 5 μg/ml showed nearly equal confining effects on FGF reactivity. The cell viability of each experimental group (named after supplemented GAGs) was compared with FGF control by 2-sample t tests. Data are mean ± SD ( n = 3). * P
    Figure Legend Snippet: GAGs restrict the diffusion gradient of FGF10. a A hydrogel cell culture system was built to mimic the ECM environment. BaF3-FGFR2b cells were embedded in hydrogel with or without GAGs and cultured in RPMI1640 media. Cultures without GAGs in the hydrogel were divided into blank control or FGF10 control according to the presence or absence of 1000 pM FGF10 in the culture medium. The experimental groups were supplemented with 1000 pM FGF10 in the culture medium plus 1 μg/ml, 3 μg/ml, or 5 μg/ml of GAGs in the hydrogel. After 48 h of culture, the FGF activity of each group was calculated based on the viability of BaF3-FGFR2b cells (see Additional File 10: Fig. S10 ). b Cultures with GAGs in the hydrogel without FGF10 in the medium showed no significant differences from the blank control, indicating that GAGs alone did not affect the viability of BaF3 cells. c , d GAGs at 1 μg/ml and 3 μg/ml concentrations displayed differential config effects on FGF reactivity. e GAGs at 5 μg/ml showed nearly equal confining effects on FGF reactivity. The cell viability of each experimental group (named after supplemented GAGs) was compared with FGF control by 2-sample t tests. Data are mean ± SD ( n = 3). * P

    Techniques Used: Diffusion-based Assay, Cell Culture, Activity Assay

    11) Product Images from "A Novel, Fully Human Anti–fucosyl-GM1 Antibody Demonstrates Potent In Vitro and In Vivo Antitumor Activity in Preclinical Models of Small Cell Lung Cancer"

    Article Title: A Novel, Fully Human Anti–fucosyl-GM1 Antibody Demonstrates Potent In Vitro and In Vivo Antitumor Activity in Preclinical Models of Small Cell Lung Cancer

    Journal: Clinical cancer research : an official journal of the American Association for Cancer Research

    doi: 10.1158/1078-0432.CCR-18-0018

    BMS-986012 efficacy is enhanced by anti-CD137 antibody. A, BMS-986012 induces NK-cell activation in the presence of DMS79 target cells. Negatively selected human NK cells and DMS79 target cells were cocultured 1:1 for 24 hours in RPMI1640 plus 10% FBS without IL2 with serially diluted BMS-986012 or isotype control antibody. Expression of CD137 (left) and CD107a (middle) was determined by flow cytometry. IFNγ secreted into culture supernatants was determined by ELISA (right). Data are representative of three independent experiments. B, Mice were dosed every 3 to 4 days with 3 mg/kg BMS-986012 beginning on day 7 postimplantation or 1 mg/kg anti-mouse CD137 beginning day 8 posttransplantation to assess single-agent efficacy. To evaluate combined activity, mice were dosed with 3 mg/kg BMS-986012 every 3 to 4 days intraperitoneally beginning on day 7 after tumor implantation and either 0.1 or 0.3 mg/kg anti-mouse CD137 intraperitoneally 1 day after each dose of BMS-986012 beginning on day 8 post implantation. Data are representative of two independent experiments.
    Figure Legend Snippet: BMS-986012 efficacy is enhanced by anti-CD137 antibody. A, BMS-986012 induces NK-cell activation in the presence of DMS79 target cells. Negatively selected human NK cells and DMS79 target cells were cocultured 1:1 for 24 hours in RPMI1640 plus 10% FBS without IL2 with serially diluted BMS-986012 or isotype control antibody. Expression of CD137 (left) and CD107a (middle) was determined by flow cytometry. IFNγ secreted into culture supernatants was determined by ELISA (right). Data are representative of three independent experiments. B, Mice were dosed every 3 to 4 days with 3 mg/kg BMS-986012 beginning on day 7 postimplantation or 1 mg/kg anti-mouse CD137 beginning day 8 posttransplantation to assess single-agent efficacy. To evaluate combined activity, mice were dosed with 3 mg/kg BMS-986012 every 3 to 4 days intraperitoneally beginning on day 7 after tumor implantation and either 0.1 or 0.3 mg/kg anti-mouse CD137 intraperitoneally 1 day after each dose of BMS-986012 beginning on day 8 post implantation. Data are representative of two independent experiments.

    Techniques Used: Activation Assay, Expressing, Flow Cytometry, Cytometry, Enzyme-linked Immunosorbent Assay, Mouse Assay, Activity Assay, Tumor Implantation

    12) Product Images from "CXCR6 regulates the recruitment of pro-inflammatory IL-17A-producing T cells into atherosclerotic aortas"

    Article Title: CXCR6 regulates the recruitment of pro-inflammatory IL-17A-producing T cells into atherosclerotic aortas

    Journal: International Immunology

    doi: 10.1093/intimm/dxv068

    T h17 cells recovered from atherosclerotic mouse spleens chemotactically migrate toward CXCL16. Forty- to fifty-week-old Apoe −/− and Cxcr6 GFP/GFP Apoe −/− peripheral lymph nodes were collected and 0.3–0.4×10 6 cells were seeded into transwells containing complete RPMI1640 supplemented with BSA, 100ng ml –1 CXCL16, 300ng ml –1 CXCL16 or 1000ng ml –1 CCL20 for cell migration experiments. The cells were allowed to migrate for 3h before the cells in the bottom well were collected for re-stimulation and intracellular cytokine staining. The results depict IL-17A + ). (A) Fold Cxcr6 GFP/+ Apoe −/− IL-17A + T-cell transwell migration. (B) Fold Cxcr6 GFP/GFP Apoe −/− IL-17A + T-cell transwell migration. The results are shown as the mean ± SEM and are representative of three technical replicates from three to four independent experiments. The data are expressed as the mean ± SD of triplicate wells. * P
    Figure Legend Snippet: T h17 cells recovered from atherosclerotic mouse spleens chemotactically migrate toward CXCL16. Forty- to fifty-week-old Apoe −/− and Cxcr6 GFP/GFP Apoe −/− peripheral lymph nodes were collected and 0.3–0.4×10 6 cells were seeded into transwells containing complete RPMI1640 supplemented with BSA, 100ng ml –1 CXCL16, 300ng ml –1 CXCL16 or 1000ng ml –1 CCL20 for cell migration experiments. The cells were allowed to migrate for 3h before the cells in the bottom well were collected for re-stimulation and intracellular cytokine staining. The results depict IL-17A + ). (A) Fold Cxcr6 GFP/+ Apoe −/− IL-17A + T-cell transwell migration. (B) Fold Cxcr6 GFP/GFP Apoe −/− IL-17A + T-cell transwell migration. The results are shown as the mean ± SEM and are representative of three technical replicates from three to four independent experiments. The data are expressed as the mean ± SD of triplicate wells. * P

    Techniques Used: Migration, Staining

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    Incubation:

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    Cell Culture:

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    Article Title: Neutrophil Elastase Subverts the Immune Response by Cleaving Toll-Like Receptors and Cytokines in Pneumococcal Pneumonia
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    Infection:

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    Mouse Assay:

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