Structured Review

FUJIFILM ionomycin
ftz-f1 knockdown causes defects in ovulation and egg morphology. ( A–C ) Ftz-f1 protein (green in A-C) in stage-12 egg chambers from control ( A ), ftz-f1 RNAi1 ( B ), and ftz-f1R NAi2 ( C ) females with Vm26Aa-Gal4, UAS-dcr2; Oamb-RFP . The insets are higher magnification of Ftz-f1 expression in squared areas. ( D–E ) Quantification of egg laying ( D ) and mature follicles in females post egg laying ( E ) in control or ftz-f1 RNAi females with Vm26Aa-Gal4, UAS-dcr2; 47A04-LexA, lexAop2-6XGFP . The number of females is noted above each bar. ( F ) Quantification of OA- and <t>Ionomycin-induced</t> follicle rupture using mature follicles isolated from control or ftz-f1 RNAi females. Mature follicles were isolated according to 47A04-lexA > 6 XGFP expression. The number of mature follicles analyzed is noted above each bar. ( G–I ) Representative images show follicles from control ( G ), ftz-f1 RNA1i ( H ), and ftz-f1 RNAi2 ( I ) females after 3 hr culture with OA. Follicles were isolated according to Oamb-RFP expression (red). Ruptured follicles are marked by arrowheads. ( J–L ) Representative images show follicles from control ( J ), ftz-f1 RNA1i ( K ), and ftz-f1 RNAi2 ( L ) females after 3 hr culture with OA. Follicles were isolated according to 47A04-lexA > 6 XGFP expression (green). Ruptured follicles are marked by arrowheads. 6XGFP forms puncta inside follicle cells. ( M–O ) Representative DIC images show dorsal appendage morphology in control ( M ), ftz-f1 RNA1i ( N ), and ftz-f1 RNAi2 ( O ) stage-14 egg chambers. Blue arrowheads indicate stunted dorsal appendage formation. ***p
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Images

1) Product Images from "Nuclear receptor Ftz-f1 promotes follicle maturation and ovulation partly via bHLH/PAS transcription factor Sim"

Article Title: Nuclear receptor Ftz-f1 promotes follicle maturation and ovulation partly via bHLH/PAS transcription factor Sim

Journal: eLife

doi: 10.7554/eLife.54568

ftz-f1 knockdown causes defects in ovulation and egg morphology. ( A–C ) Ftz-f1 protein (green in A-C) in stage-12 egg chambers from control ( A ), ftz-f1 RNAi1 ( B ), and ftz-f1R NAi2 ( C ) females with Vm26Aa-Gal4, UAS-dcr2; Oamb-RFP . The insets are higher magnification of Ftz-f1 expression in squared areas. ( D–E ) Quantification of egg laying ( D ) and mature follicles in females post egg laying ( E ) in control or ftz-f1 RNAi females with Vm26Aa-Gal4, UAS-dcr2; 47A04-LexA, lexAop2-6XGFP . The number of females is noted above each bar. ( F ) Quantification of OA- and Ionomycin-induced follicle rupture using mature follicles isolated from control or ftz-f1 RNAi females. Mature follicles were isolated according to 47A04-lexA > 6 XGFP expression. The number of mature follicles analyzed is noted above each bar. ( G–I ) Representative images show follicles from control ( G ), ftz-f1 RNA1i ( H ), and ftz-f1 RNAi2 ( I ) females after 3 hr culture with OA. Follicles were isolated according to Oamb-RFP expression (red). Ruptured follicles are marked by arrowheads. ( J–L ) Representative images show follicles from control ( J ), ftz-f1 RNA1i ( K ), and ftz-f1 RNAi2 ( L ) females after 3 hr culture with OA. Follicles were isolated according to 47A04-lexA > 6 XGFP expression (green). Ruptured follicles are marked by arrowheads. 6XGFP forms puncta inside follicle cells. ( M–O ) Representative DIC images show dorsal appendage morphology in control ( M ), ftz-f1 RNA1i ( N ), and ftz-f1 RNAi2 ( O ) stage-14 egg chambers. Blue arrowheads indicate stunted dorsal appendage formation. ***p
Figure Legend Snippet: ftz-f1 knockdown causes defects in ovulation and egg morphology. ( A–C ) Ftz-f1 protein (green in A-C) in stage-12 egg chambers from control ( A ), ftz-f1 RNAi1 ( B ), and ftz-f1R NAi2 ( C ) females with Vm26Aa-Gal4, UAS-dcr2; Oamb-RFP . The insets are higher magnification of Ftz-f1 expression in squared areas. ( D–E ) Quantification of egg laying ( D ) and mature follicles in females post egg laying ( E ) in control or ftz-f1 RNAi females with Vm26Aa-Gal4, UAS-dcr2; 47A04-LexA, lexAop2-6XGFP . The number of females is noted above each bar. ( F ) Quantification of OA- and Ionomycin-induced follicle rupture using mature follicles isolated from control or ftz-f1 RNAi females. Mature follicles were isolated according to 47A04-lexA > 6 XGFP expression. The number of mature follicles analyzed is noted above each bar. ( G–I ) Representative images show follicles from control ( G ), ftz-f1 RNA1i ( H ), and ftz-f1 RNAi2 ( I ) females after 3 hr culture with OA. Follicles were isolated according to Oamb-RFP expression (red). Ruptured follicles are marked by arrowheads. ( J–L ) Representative images show follicles from control ( J ), ftz-f1 RNA1i ( K ), and ftz-f1 RNAi2 ( L ) females after 3 hr culture with OA. Follicles were isolated according to 47A04-lexA > 6 XGFP expression (green). Ruptured follicles are marked by arrowheads. 6XGFP forms puncta inside follicle cells. ( M–O ) Representative DIC images show dorsal appendage morphology in control ( M ), ftz-f1 RNA1i ( N ), and ftz-f1 RNAi2 ( O ) stage-14 egg chambers. Blue arrowheads indicate stunted dorsal appendage formation. ***p

Techniques Used: Expressing, Isolation

2) Product Images from "Conditional Deletion of TAK1 in T Cells Reveals a Pivotal Role of TCRαβ+ Intraepithelial Lymphocytes in Preventing Lymphopenia-Associated Colitis"

Article Title: Conditional Deletion of TAK1 in T Cells Reveals a Pivotal Role of TCRαβ+ Intraepithelial Lymphocytes in Preventing Lymphopenia-Associated Colitis

Journal: PLoS ONE

doi: 10.1371/journal.pone.0128761

Characterization of aberrant T cells in LTAC mice. (A and B) Frequencies (left) and absolute numbers (right) of TCRβ + cells of CD45 + cells from mLNs (A) or cLP (B) of WT (n = 9–10, filled circle) and LTAC mice (n = 9–11, open circle) determined by flow cytometry analysis. Horizontal bars represent mean. (C) (Left and middle plots) Flow cytometry of CD4 + and CD8α + cell subsets in TCRβ + cells. (Right panels) Frequencies of CD4 + cell subset in TCRβ + cells. The plots are representative of at least three independent experiments. mLNs (upper) or cLP (lower) of WT (n = 7–10, filled circle) and LTAC mice (n = 8–10, open circle), determined by flow cytometry analysis. Horizontal bars represent mean. (D) Genomic DNAs from the sorted T cells in each tissue indicated were subjected to PCR amplification to detect flox (= TAK1 WT) or Δ (= TAK1 KO) allele in the TAK1 genomic locus. The DNA size difference between the flox and Δ bands is shown. Specific DNAs for Cre, loaded in each well, were also shown to prove its existence in the genome. (E) Flow cytometry of intracellular cytokines in TCRβ + CD4 + cells from cLP after culture with PMA + Ionomycin for 5 hours in the presence of GolgiStop. The plots are representative of four independent experiments. (F) Frequencies of each subset of cytokine-producing cells in TCRβ + CD4 + cells in cLP of WT (n = 4, filled circle) and LTAC mice (n = 4, open circle), calculated by flow cytometry analysis from (E). Horizontal bars represent mean. (G) Flow cytometry of gut homing receptors by mLN TCRβ + CD4 + cells. The histograms are representative of at least three independent experiments. (H) Frequencies of α4β7 hi , CCR9 + and CD103 + cells of TCRβ + CD4 + cells in the mLNs of WT (n ≧ 5, filled circle) and LTAC mice (n ≧ 5, open circle), determined by flow cytometry analysis from (H). Horizontal bars represent mean. In (A), (B), (C), (F) and (H), unpaired t tests were performed. Statistical significance was indicated by * P
Figure Legend Snippet: Characterization of aberrant T cells in LTAC mice. (A and B) Frequencies (left) and absolute numbers (right) of TCRβ + cells of CD45 + cells from mLNs (A) or cLP (B) of WT (n = 9–10, filled circle) and LTAC mice (n = 9–11, open circle) determined by flow cytometry analysis. Horizontal bars represent mean. (C) (Left and middle plots) Flow cytometry of CD4 + and CD8α + cell subsets in TCRβ + cells. (Right panels) Frequencies of CD4 + cell subset in TCRβ + cells. The plots are representative of at least three independent experiments. mLNs (upper) or cLP (lower) of WT (n = 7–10, filled circle) and LTAC mice (n = 8–10, open circle), determined by flow cytometry analysis. Horizontal bars represent mean. (D) Genomic DNAs from the sorted T cells in each tissue indicated were subjected to PCR amplification to detect flox (= TAK1 WT) or Δ (= TAK1 KO) allele in the TAK1 genomic locus. The DNA size difference between the flox and Δ bands is shown. Specific DNAs for Cre, loaded in each well, were also shown to prove its existence in the genome. (E) Flow cytometry of intracellular cytokines in TCRβ + CD4 + cells from cLP after culture with PMA + Ionomycin for 5 hours in the presence of GolgiStop. The plots are representative of four independent experiments. (F) Frequencies of each subset of cytokine-producing cells in TCRβ + CD4 + cells in cLP of WT (n = 4, filled circle) and LTAC mice (n = 4, open circle), calculated by flow cytometry analysis from (E). Horizontal bars represent mean. (G) Flow cytometry of gut homing receptors by mLN TCRβ + CD4 + cells. The histograms are representative of at least three independent experiments. (H) Frequencies of α4β7 hi , CCR9 + and CD103 + cells of TCRβ + CD4 + cells in the mLNs of WT (n ≧ 5, filled circle) and LTAC mice (n ≧ 5, open circle), determined by flow cytometry analysis from (H). Horizontal bars represent mean. In (A), (B), (C), (F) and (H), unpaired t tests were performed. Statistical significance was indicated by * P

Techniques Used: Mouse Assay, Flow Cytometry, Cytometry, Polymerase Chain Reaction, Amplification

3) Product Images from "High fat diet exacerbates murine psoriatic dermatitis by increasing the number of IL-17-producing γδ T cells"

Article Title: High fat diet exacerbates murine psoriatic dermatitis by increasing the number of IL-17-producing γδ T cells

Journal: Scientific Reports

doi: 10.1038/s41598-017-14292-1

Increase of IL-17A-producing cells in the skin of HFD-fed mice in both steady and inflammatory states. ( a ) Flow cytometric analysis of IL-17A-producing cells in the whole ear skin of HFD- or ND-fed mice in the steady state. The single cell suspensions were stimulated with phorbol myristate acetate (PMA) and ionomycin in the presence of brefeldin A for 3 h before intracellular staining. Left panels indicate cells gated on CD45 + , and right panels indicate cells gated on IL-17A + in the left panels. ( b – e ) Statistical analysis of the number of total IL-17A-producing cells ( b ) and the percentage ( c ) and number/composition ( d , e ) of IL-17A-producing cells. Samples were collected at 24 h after the last IMQ treatment, and the ear skin was digested with collagenase in the presence or absence of brefeldin A. Single cell suspensions were either stimulated with PMA and ionomycin in the presence of brefeldin A for 3 h before intracellular staining ( b – d ) or directly subjected to intracellular staining ( b , e ). ( f ) Fold induction of Il23a and Tnfa mRNA in the whole ear skin of ND- or HFD-fed mice in the steady state, as analyzed by quantitative RT-PCR. Results are presented relative to those of ND. The average mRNA expression level in ND-fed mice is set as 1. ( g ) Flow cytometric analysis of the ratio of Ki67 + TCRγδ + cells in the whole ear skin in the steady state. Results are expressed as the mean ± SEM. p -values were obtained by Mann-Whitney-U-test ( b , f , g ) and one-way ANOVA ( d , e ). * p ≤ 0.05. Data are from one experiment, representative of four independent experiments with three to four mice ( a ), two experiments with four mice ( g ). Data are pooled from two experiments with three to four mice ( b – e ), three experiments with three mice ( f ).
Figure Legend Snippet: Increase of IL-17A-producing cells in the skin of HFD-fed mice in both steady and inflammatory states. ( a ) Flow cytometric analysis of IL-17A-producing cells in the whole ear skin of HFD- or ND-fed mice in the steady state. The single cell suspensions were stimulated with phorbol myristate acetate (PMA) and ionomycin in the presence of brefeldin A for 3 h before intracellular staining. Left panels indicate cells gated on CD45 + , and right panels indicate cells gated on IL-17A + in the left panels. ( b – e ) Statistical analysis of the number of total IL-17A-producing cells ( b ) and the percentage ( c ) and number/composition ( d , e ) of IL-17A-producing cells. Samples were collected at 24 h after the last IMQ treatment, and the ear skin was digested with collagenase in the presence or absence of brefeldin A. Single cell suspensions were either stimulated with PMA and ionomycin in the presence of brefeldin A for 3 h before intracellular staining ( b – d ) or directly subjected to intracellular staining ( b , e ). ( f ) Fold induction of Il23a and Tnfa mRNA in the whole ear skin of ND- or HFD-fed mice in the steady state, as analyzed by quantitative RT-PCR. Results are presented relative to those of ND. The average mRNA expression level in ND-fed mice is set as 1. ( g ) Flow cytometric analysis of the ratio of Ki67 + TCRγδ + cells in the whole ear skin in the steady state. Results are expressed as the mean ± SEM. p -values were obtained by Mann-Whitney-U-test ( b , f , g ) and one-way ANOVA ( d , e ). * p ≤ 0.05. Data are from one experiment, representative of four independent experiments with three to four mice ( a ), two experiments with four mice ( g ). Data are pooled from two experiments with three to four mice ( b – e ), three experiments with three mice ( f ).

Techniques Used: Mouse Assay, Flow Cytometry, Staining, Quantitative RT-PCR, Expressing, MANN-WHITNEY

4) Product Images from "Intracellular calcium signal at the leading edge regulates mesodermal sheet migration during Xenopus gastrulation"

Article Title: Intracellular calcium signal at the leading edge regulates mesodermal sheet migration during Xenopus gastrulation

Journal: Scientific Reports

doi: 10.1038/s41598-018-20747-w

Ca 2+ signalling regulates Rac1 activity. ( a ) Rac1 activity after Ionomycin (Iono.) treatment for 3 minutes and 2 hours. DMZ explants were dissected and treated with Ionomycin. DMSO was used as a control. ( b ) Rac1 activity after Ionomycin treatment for 3 minutes. The active Rac1 intensity with Ionomycin was normalized to the intensity in DMSO-treated samples. Each western sample was prepared from 35–40 DMZ explants. Error bars indicate s.e. ± Student’s t-test, ns: No significance. ( c ) Rac1 activity after Ionomycin treatment for 2 hours. Student’s t-test, **P
Figure Legend Snippet: Ca 2+ signalling regulates Rac1 activity. ( a ) Rac1 activity after Ionomycin (Iono.) treatment for 3 minutes and 2 hours. DMZ explants were dissected and treated with Ionomycin. DMSO was used as a control. ( b ) Rac1 activity after Ionomycin treatment for 3 minutes. The active Rac1 intensity with Ionomycin was normalized to the intensity in DMSO-treated samples. Each western sample was prepared from 35–40 DMZ explants. Error bars indicate s.e. ± Student’s t-test, ns: No significance. ( c ) Rac1 activity after Ionomycin treatment for 2 hours. Student’s t-test, **P

Techniques Used: Activity Assay, Western Blot

Intracellular Ca 2+ signalling regulates migration activity in the LEM. ( a ) Snapshots from time-lapse imaging of the Ca 2+ dynamics in DMZ explants treated with Ionomycin (2.5 μM). Upper panel: mRFP. Lower panel: FRET ratio of yellow cameleon-nano converted to pseudocolours (bar at right). ( b ) LEM migration into open space in Ionomycin- (2.5 μM) and DMSO-treated DMZ explants monitored by mRFP. ( c ) Relative migration distance in DMSO- and Ionomycin-treated DMZ explants. The migration distance was normalized to the migration distance of DMSO-treated DMZ. DMSO: n = 7 embryos. Ionomycin (Iono.): n = 8 embryos. Error bars indicate s.e. ± Student’s t-test, *P
Figure Legend Snippet: Intracellular Ca 2+ signalling regulates migration activity in the LEM. ( a ) Snapshots from time-lapse imaging of the Ca 2+ dynamics in DMZ explants treated with Ionomycin (2.5 μM). Upper panel: mRFP. Lower panel: FRET ratio of yellow cameleon-nano converted to pseudocolours (bar at right). ( b ) LEM migration into open space in Ionomycin- (2.5 μM) and DMSO-treated DMZ explants monitored by mRFP. ( c ) Relative migration distance in DMSO- and Ionomycin-treated DMZ explants. The migration distance was normalized to the migration distance of DMSO-treated DMZ. DMSO: n = 7 embryos. Ionomycin (Iono.): n = 8 embryos. Error bars indicate s.e. ± Student’s t-test, *P

Techniques Used: Migration, Activity Assay, Imaging

5) Product Images from "Intracellular calcium signal at the leading edge regulates mesodermal sheet migration during Xenopus gastrulation"

Article Title: Intracellular calcium signal at the leading edge regulates mesodermal sheet migration during Xenopus gastrulation

Journal: Scientific Reports

doi: 10.1038/s41598-018-20747-w

Ca 2+ signalling regulates Rac1 activity. ( a ) Rac1 activity after Ionomycin (Iono.) treatment for 3 minutes and 2 hours. DMZ explants were dissected and treated with Ionomycin. DMSO was used as a control. ( b ) Rac1 activity after Ionomycin treatment for 3 minutes. The active Rac1 intensity with Ionomycin was normalized to the intensity in DMSO-treated samples. Each western sample was prepared from 35–40 DMZ explants. Error bars indicate s.e. ± Student’s t-test, ns: No significance. ( c ) Rac1 activity after Ionomycin treatment for 2 hours. Student’s t-test, **P
Figure Legend Snippet: Ca 2+ signalling regulates Rac1 activity. ( a ) Rac1 activity after Ionomycin (Iono.) treatment for 3 minutes and 2 hours. DMZ explants were dissected and treated with Ionomycin. DMSO was used as a control. ( b ) Rac1 activity after Ionomycin treatment for 3 minutes. The active Rac1 intensity with Ionomycin was normalized to the intensity in DMSO-treated samples. Each western sample was prepared from 35–40 DMZ explants. Error bars indicate s.e. ± Student’s t-test, ns: No significance. ( c ) Rac1 activity after Ionomycin treatment for 2 hours. Student’s t-test, **P

Techniques Used: Activity Assay, Western Blot

Intracellular Ca 2+ signalling regulates migration activity in the LEM. ( a ) Snapshots from time-lapse imaging of the Ca 2+ dynamics in DMZ explants treated with Ionomycin (2.5 μM). Upper panel: mRFP. Lower panel: FRET ratio of yellow cameleon-nano converted to pseudocolours (bar at right). ( b ) LEM migration into open space in Ionomycin- (2.5 μM) and DMSO-treated DMZ explants monitored by mRFP. ( c ) Relative migration distance in DMSO- and Ionomycin-treated DMZ explants. The migration distance was normalized to the migration distance of DMSO-treated DMZ. DMSO: n = 7 embryos. Ionomycin (Iono.): n = 8 embryos. Error bars indicate s.e. ± Student’s t-test, *P
Figure Legend Snippet: Intracellular Ca 2+ signalling regulates migration activity in the LEM. ( a ) Snapshots from time-lapse imaging of the Ca 2+ dynamics in DMZ explants treated with Ionomycin (2.5 μM). Upper panel: mRFP. Lower panel: FRET ratio of yellow cameleon-nano converted to pseudocolours (bar at right). ( b ) LEM migration into open space in Ionomycin- (2.5 μM) and DMSO-treated DMZ explants monitored by mRFP. ( c ) Relative migration distance in DMSO- and Ionomycin-treated DMZ explants. The migration distance was normalized to the migration distance of DMSO-treated DMZ. DMSO: n = 7 embryos. Ionomycin (Iono.): n = 8 embryos. Error bars indicate s.e. ± Student’s t-test, *P

Techniques Used: Migration, Activity Assay, Imaging

6) Product Images from "Hypoxia-induced sensitisation of TRPA1 in painful dysesthesia evoked by transient hindlimb ischemia/reperfusion in mice"

Article Title: Hypoxia-induced sensitisation of TRPA1 in painful dysesthesia evoked by transient hindlimb ischemia/reperfusion in mice

Journal: Scientific Reports

doi: 10.1038/srep23261

Effects of hypoxia on H 2 O 2 -evoked TRPA1 activation in hTRPA1-expressing cells. HEK293 cells transfected with vector ( a , d ; mock) or hTRPA1 cDNA ( b , c , e ) were pretreated with normoxia (160 mmHg) or hypoxia (100, 90, or 80 mmHg) for 30 min, and then exposed to H 2 O 2 (10 μM) for 5 min, as shown in Supplementary Fig. S5 . ( a–c ) Representative images of [Ca 2+ ] i changes are shown as F 340 /F 380 ratio. F 340 /F 380 ratio in each cell is shown as pale-coloured lines, and the average of F 340 /F 380 ratio is shown as a black solid line. ( d,e ) Statistical analyses of H 2 O 2 -evoked [Ca 2+ ] i increases were calculated as the average of the maximal ΔRatio during the 5 min H 2 O 2 application period (33–38 min) obtained from 5–6 independent experiments, as described in Materials and Methods. AITC, allyl isothiocyanate, 100 μM; Iono, ionomycin, 3 μM. ** P
Figure Legend Snippet: Effects of hypoxia on H 2 O 2 -evoked TRPA1 activation in hTRPA1-expressing cells. HEK293 cells transfected with vector ( a , d ; mock) or hTRPA1 cDNA ( b , c , e ) were pretreated with normoxia (160 mmHg) or hypoxia (100, 90, or 80 mmHg) for 30 min, and then exposed to H 2 O 2 (10 μM) for 5 min, as shown in Supplementary Fig. S5 . ( a–c ) Representative images of [Ca 2+ ] i changes are shown as F 340 /F 380 ratio. F 340 /F 380 ratio in each cell is shown as pale-coloured lines, and the average of F 340 /F 380 ratio is shown as a black solid line. ( d,e ) Statistical analyses of H 2 O 2 -evoked [Ca 2+ ] i increases were calculated as the average of the maximal ΔRatio during the 5 min H 2 O 2 application period (33–38 min) obtained from 5–6 independent experiments, as described in Materials and Methods. AITC, allyl isothiocyanate, 100 μM; Iono, ionomycin, 3 μM. ** P

Techniques Used: Activation Assay, Expressing, Transfection, Plasmid Preparation

7) Product Images from "Early pathogenesis of Duchenne muscular dystrophy modelled in patient-derived human induced pluripotent stem cells"

Article Title: Early pathogenesis of Duchenne muscular dystrophy modelled in patient-derived human induced pluripotent stem cells

Journal: Scientific Reports

doi: 10.1038/srep12831

Ca 2+ influx induces prominent skeletal muscle cellular damage in DMD-Myocytes. ( a ) Experimental design of the CK assay. ( b ) CK activity (IU/L) at the baseline condition of Control- and DMD-Myocytes, measured after 10 min of incubation at 37 °C. n = 6 with three independent experiments conducted for each assay. ( c ) Relative fold change in the CK value in Control- and DMD-Myocytes upon ionomycin addition, normalised against CK values for DMSO (control) addition. Two-way analysis of variance (ANOVA) revealed a significant difference in the rate of change between Control- and DMD- Myocytes in response to ionomycin treatment based on Scheffe’s test. **P
Figure Legend Snippet: Ca 2+ influx induces prominent skeletal muscle cellular damage in DMD-Myocytes. ( a ) Experimental design of the CK assay. ( b ) CK activity (IU/L) at the baseline condition of Control- and DMD-Myocytes, measured after 10 min of incubation at 37 °C. n = 6 with three independent experiments conducted for each assay. ( c ) Relative fold change in the CK value in Control- and DMD-Myocytes upon ionomycin addition, normalised against CK values for DMSO (control) addition. Two-way analysis of variance (ANOVA) revealed a significant difference in the rate of change between Control- and DMD- Myocytes in response to ionomycin treatment based on Scheffe’s test. **P

Techniques Used: Activity Assay, Incubation

8) Product Images from "Streptococcus thermophilus ST28 Ameliorates Colitis in Mice Partially by Suppression of Inflammatory Th17 Cells"

Article Title: Streptococcus thermophilus ST28 Ameliorates Colitis in Mice Partially by Suppression of Inflammatory Th17 Cells

Journal: Journal of Biomedicine and Biotechnology

doi: 10.1155/2011/378417

Effects of S. thermophilus ST28 on percentages of Th17 (a) and inflammatory DC (b) in LPLs from DSS-induced colitis mice. (a) After the experiments described in Figure 3 , LPLs from DSS-induced mice orally administered ST28 or ATCC 19258 were incubated with phorbol-12-myristate-13-acetate, ionomycin, and brefeldin A and applied to flow cytometry to measure intracellular IL-17. (b) Separately, flow cytometric analysis of surface marker CD86 was performed using freshly prepared LPLs. LPLs from untreated DSS-induced colitis mice (−) and healthy control mice (Cont) were also assayed. ND, not detected.
Figure Legend Snippet: Effects of S. thermophilus ST28 on percentages of Th17 (a) and inflammatory DC (b) in LPLs from DSS-induced colitis mice. (a) After the experiments described in Figure 3 , LPLs from DSS-induced mice orally administered ST28 or ATCC 19258 were incubated with phorbol-12-myristate-13-acetate, ionomycin, and brefeldin A and applied to flow cytometry to measure intracellular IL-17. (b) Separately, flow cytometric analysis of surface marker CD86 was performed using freshly prepared LPLs. LPLs from untreated DSS-induced colitis mice (−) and healthy control mice (Cont) were also assayed. ND, not detected.

Techniques Used: Mouse Assay, Incubation, Flow Cytometry, Cytometry, Marker

9) Product Images from "Seihai-to (TJ-90)-Induced Activation of Airway Ciliary Beatings of Mice: Ca2+ Modulation of cAMP-Stimulated Ciliary Beatings via PDE1"

Article Title: Seihai-to (TJ-90)-Induced Activation of Airway Ciliary Beatings of Mice: Ca2+ Modulation of cAMP-Stimulated Ciliary Beatings via PDE1

Journal: International Journal of Molecular Sciences

doi: 10.3390/ijms19030658

Effects of ionomycin (500 nM, IM) on CBF increase and CBA increase stimulated by procaterol (100 pM). The stimulation with 100 pM procaterol increased and sustained CBF and CBA by ~35%. The further addition of 500 nM IM decreased CBF (7/10 cells) or did not change CBF (3/10 cells), although it only sustained the CBA increase. ( A ) Cells were first stimulated with 100 pM procaterol for 5 min, and then, further with 500 nM IM. The addition of 500 nM IM decreased the CBF ratio gradually ( n = 4), but not CBA ( n = 3). The incidence was 7/10 cells; ( B ) The further addition of 500 nM IM did not change the CBF ratio ( n = 3), and also the CBA ratio ( n = 3). The incidence was 3/10 cells. * shows control values and † shows the values just before the addition of IM (A).
Figure Legend Snippet: Effects of ionomycin (500 nM, IM) on CBF increase and CBA increase stimulated by procaterol (100 pM). The stimulation with 100 pM procaterol increased and sustained CBF and CBA by ~35%. The further addition of 500 nM IM decreased CBF (7/10 cells) or did not change CBF (3/10 cells), although it only sustained the CBA increase. ( A ) Cells were first stimulated with 100 pM procaterol for 5 min, and then, further with 500 nM IM. The addition of 500 nM IM decreased the CBF ratio gradually ( n = 4), but not CBA ( n = 3). The incidence was 7/10 cells; ( B ) The further addition of 500 nM IM did not change the CBF ratio ( n = 3), and also the CBA ratio ( n = 3). The incidence was 3/10 cells. * shows control values and † shows the values just before the addition of IM (A).

Techniques Used: Crocin Bleaching Assay

Related Articles

Isolation:

Article Title: Nuclear receptor Ftz-f1 promotes follicle maturation and ovulation partly via bHLH/PAS transcription factor Sim
Article Snippet: .. Five mature follicles were isolated and placed in each well of a 96-well plate with 100 μl of Grace’s insect medium containing either 20 μM OA or 2 μM ionomycin and 200 μM of L-012 (Wako Chemicals). .. Plates were placed in a CLARIOstar microplate reader (BMG Labtech) for luminescence reading for 60 min.

Cell Culture:

Article Title: Early pathogenesis of Duchenne muscular dystrophy modelled in patient-derived human induced pluripotent stem cells
Article Snippet: .. Collected supernatants were supplemented with protease inhibitor cocktail (Nacalai Tesque) at 1:100 dilution and CK activity was measured by Oriental Yeast Co., LTD. Ca2+ influx analysis was performed by adding 6.25 μM ionomycin (Wako) or ionomycin and 1 μM ruthenium red (Sigma) to cultured cells. .. Ca2+ uptake was visualised with Fluo-8 and measured with the BZ9000 system.

Article Title: Conditional Deletion of TAK1 in T Cells Reveals a Pivotal Role of TCRαβ+ Intraepithelial Lymphocytes in Preventing Lymphopenia-Associated Colitis
Article Snippet: .. For intracellular cytokine staining, cells were cultured with 50 ng/ml phorbol 12-myristate 13-acetate (PMA) (Wako) plus 500 ng/ml ionomycin (Wako) for 5 hours in the presence of GolgiStop (BD Bioscience). .. After staining with antibodies to cell surface molecules, cells were fixed and permeabilized in Cytofix/Cytoperm buffer (BD Bioscience), followed by intracellular cytokine staining in Perm/Wash buffer (BD Bioscience).

Incubation:

Article Title: Streptococcus thermophilus ST28 Ameliorates Colitis in Mice Partially by Suppression of Inflammatory Th17 Cells
Article Snippet: .. Flow Cytometric Analysis of Splenocytes The splenocytes (1 × 106 cells/mL) were incubated at 37°C for 4 hours in RPMI 1640 medium containing 0.5 μ g/mL phorbol-12-myristate-13-acetate (MP Biomedicals, Aurora, Ohio, USA), 1 μ g/mL ionomycin (Wako Pure Chemical Industries, Osaka, Japan), and 3 μ g/mL brefeldin-A (Wako Pure Chemical Industries). .. Then, anti-mouse CD32/16 antibody was added and incubated 4°C for 30 min. For analysis of surface markers, fluorescein isothiocyanate (FITC) anti-mouse CD11c antibody (BioLegend, San Diego, Calif, USA), allophycocyanin (APC) anti-mouse CD11b antibody (eBioscience, San Diego, Calif, USA), or APC anti-mouse CD4 antibody (eBioscience) was added to the cell suspension, which was then incubated in the dark at 4°C for 30 min. For isotype controls, FITC-Armenian hamster IgG, APC rat IgG2aκ (BioLegend), or APC Rat IgG2bκ (eBioscience) was used.

Activity Assay:

Article Title: Early pathogenesis of Duchenne muscular dystrophy modelled in patient-derived human induced pluripotent stem cells
Article Snippet: .. Collected supernatants were supplemented with protease inhibitor cocktail (Nacalai Tesque) at 1:100 dilution and CK activity was measured by Oriental Yeast Co., LTD. Ca2+ influx analysis was performed by adding 6.25 μM ionomycin (Wako) or ionomycin and 1 μM ruthenium red (Sigma) to cultured cells. .. Ca2+ uptake was visualised with Fluo-8 and measured with the BZ9000 system.

Protease Inhibitor:

Article Title: Early pathogenesis of Duchenne muscular dystrophy modelled in patient-derived human induced pluripotent stem cells
Article Snippet: .. Collected supernatants were supplemented with protease inhibitor cocktail (Nacalai Tesque) at 1:100 dilution and CK activity was measured by Oriental Yeast Co., LTD. Ca2+ influx analysis was performed by adding 6.25 μM ionomycin (Wako) or ionomycin and 1 μM ruthenium red (Sigma) to cultured cells. .. Ca2+ uptake was visualised with Fluo-8 and measured with the BZ9000 system.

Staining:

Article Title: High fat diet exacerbates murine psoriatic dermatitis by increasing the number of IL-17-producing γδ T cells
Article Snippet: .. For intracellular staining, cells were stimulated for 3 h with 50 ng/ml PMA (phorbol myristate acetate; Sigma-Aldrich, St Louis, MO) and 1 μg/ml ionomycin (Wako, Osaka, Japan) in the presence of 10 μg/ml brefeldin A (Sigma-Aldrich) or put 10 μg/ml brefeldin A in the collagenase solution, and then fixed and permeabilized with Cytofix/Cytoperm buffer (BD Biosciences). .. Flow cytometry was performed using LSRFortessa (BD Biosciences) and analyzed with FlowJo (TreeStar, San Carlos, CA).

Article Title: Conditional Deletion of TAK1 in T Cells Reveals a Pivotal Role of TCRαβ+ Intraepithelial Lymphocytes in Preventing Lymphopenia-Associated Colitis
Article Snippet: .. For intracellular cytokine staining, cells were cultured with 50 ng/ml phorbol 12-myristate 13-acetate (PMA) (Wako) plus 500 ng/ml ionomycin (Wako) for 5 hours in the presence of GolgiStop (BD Bioscience). .. After staining with antibodies to cell surface molecules, cells were fixed and permeabilized in Cytofix/Cytoperm buffer (BD Bioscience), followed by intracellular cytokine staining in Perm/Wash buffer (BD Bioscience).

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  • 92
    FUJIFILM dnase i
    Dnase I, supplied by FUJIFILM, used in various techniques. Bioz Stars score: 92/100, based on 5 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/dnase i/product/FUJIFILM
    Average 92 stars, based on 5 article reviews
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    dnase i - by Bioz Stars, 2020-09
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