Structured Review

Alomone Labs ionomycin
Specific F-actin structures in the sperm head did not change as a consequence of the AR initiation. Sperm were loaded with SiR-actin (green) and FM4-64 (magenta) and attached to concanavalin A-coated slides for imaging using TIRF microscopy. Following image acquisition, the ROIs indicated in A and D were analyzed. Representative image sequences of sperm stimulated with <t>ionomycin</t> (addition indicated by black arrow) that initially possessed the F-actin structure corresponding to the perforatorium (B), ventral (D; i) or neck (D; ii) region. The corresponding fluorescence traces (C and F) of the indicated ROIs (for SiR-actin) and the whole sperm head (for FM4-64) (A and D) are shown on the right. Analysis of these traces demonstrates that the depolymerization did not occur before or after the initiation of the AR (C and F) ( n =6. 62 cells analyzed).
Ionomycin, supplied by Alomone Labs, used in various techniques. Bioz Stars score: 91/100, based on 3 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/result/ionomycin/product/Alomone Labs
Average 91 stars, based on 3 article reviews
Price from $9.99 to $1999.99
ionomycin - by Bioz Stars, 2020-09
91/100 stars

Images

1) Product Images from "Super-resolution imaging of live sperm reveals dynamic changes of the actin cytoskeleton during acrosomal exocytosis"

Article Title: Super-resolution imaging of live sperm reveals dynamic changes of the actin cytoskeleton during acrosomal exocytosis

Journal: Journal of Cell Science

doi: 10.1242/jcs.218958

Specific F-actin structures in the sperm head did not change as a consequence of the AR initiation. Sperm were loaded with SiR-actin (green) and FM4-64 (magenta) and attached to concanavalin A-coated slides for imaging using TIRF microscopy. Following image acquisition, the ROIs indicated in A and D were analyzed. Representative image sequences of sperm stimulated with ionomycin (addition indicated by black arrow) that initially possessed the F-actin structure corresponding to the perforatorium (B), ventral (D; i) or neck (D; ii) region. The corresponding fluorescence traces (C and F) of the indicated ROIs (for SiR-actin) and the whole sperm head (for FM4-64) (A and D) are shown on the right. Analysis of these traces demonstrates that the depolymerization did not occur before or after the initiation of the AR (C and F) ( n =6. 62 cells analyzed).
Figure Legend Snippet: Specific F-actin structures in the sperm head did not change as a consequence of the AR initiation. Sperm were loaded with SiR-actin (green) and FM4-64 (magenta) and attached to concanavalin A-coated slides for imaging using TIRF microscopy. Following image acquisition, the ROIs indicated in A and D were analyzed. Representative image sequences of sperm stimulated with ionomycin (addition indicated by black arrow) that initially possessed the F-actin structure corresponding to the perforatorium (B), ventral (D; i) or neck (D; ii) region. The corresponding fluorescence traces (C and F) of the indicated ROIs (for SiR-actin) and the whole sperm head (for FM4-64) (A and D) are shown on the right. Analysis of these traces demonstrates that the depolymerization did not occur before or after the initiation of the AR (C and F) ( n =6. 62 cells analyzed).

Techniques Used: Imaging, Microscopy, Fluorescence

Specific dynamic changes in the actin cytoskeleton after initiation of acrosomal exocytosis. (A) Sperm labeled with SiR-actin (green) and FM4-64 (magenta). The ROIs used for the analysis, corresponding to the upper acrosome region (for SiR-actin) and the whole sperm head (for FM4-64), are depicted. (B) Representative sequence of images of a sperm loaded with SiR-actin and FM4-64 that displayed a decrease in SiR-actin fluorescence in the upper acrosome region after stimulation with 10†µM ionomycin (addition indicated by black arrow). ″ represents time in seconds. (C) The resulting fluorescence traces revealed that the loss of F-actin occurred after the initiation of the AR as judged by the increase in FM4-64 fluorescence ( n .
Figure Legend Snippet: Specific dynamic changes in the actin cytoskeleton after initiation of acrosomal exocytosis. (A) Sperm labeled with SiR-actin (green) and FM4-64 (magenta). The ROIs used for the analysis, corresponding to the upper acrosome region (for SiR-actin) and the whole sperm head (for FM4-64), are depicted. (B) Representative sequence of images of a sperm loaded with SiR-actin and FM4-64 that displayed a decrease in SiR-actin fluorescence in the upper acrosome region after stimulation with 10†µM ionomycin (addition indicated by black arrow). ″ represents time in seconds. (C) The resulting fluorescence traces revealed that the loss of F-actin occurred after the initiation of the AR as judged by the increase in FM4-64 fluorescence ( n .

Techniques Used: Labeling, Sequencing, Fluorescence

2) Product Images from "Balance of calcineurin A? and CDK5 activities sets release probability at nerve terminals"

Article Title: Balance of calcineurin A? and CDK5 activities sets release probability at nerve terminals

Journal: The Journal of neuroscience : the official journal of the Society for Neuroscience

doi: 10.1523/JNEUROSCI.4288-12.2013

CNAα and CDK5 gate silent presynaptic boutons by controlling Ca 2+ influx (A) Representative single-AP GCaMP3 difference image (ΔF 1AP ) in WT neurons in the absence or presence of CSA (Top) or Ros (Bottom). Ionomycin treatment (right) reveals all presynaptic boutons. White circles indicate boutons that were silenced after treatment with CSA (Top) or unsilenced after treatment of Ros (Bottom). Scale bar represents 5 μm. (B) Corresponding traces of silenced (white circle in A top) and unsilenced boutons (white circle in A bottom) with or without treatment of CSA or Ros. (C) Histogram of the fraction of silent and responsive boutons measured by GCaMP3 in WT with or without CSA or Roscovitine: WT silent = 0.17 ± 0.04, WT responsive = 0.83 ± 0.04 (n= 25), WT+CSA silent = 0.46 ± 0.09, WT+CSA responsive = 0.54 ± 0.09 (n= 9), WT+Ros silent = 0.07 ± 0.02, WT+Ros responsive = 0.93 ± 0.02 (n= 16). (D) Histogram of the proportion of silent and responsive boutons measured by MgGreen in WT with or without CSA or Ros. The division of silent or responsive boutons was based on a signal to noise ratio of 1 (peak versus SD of baseline) for the single-AP response: WT silent = 0.14 ± 0.03, WT responsive = 0.86 ± 0.03 (n=19), WT+CSA silent = 0.36 ± 0.11, WT+CSA responsive = 0.64 ± 0.11 (n=7), WT+Ros silent = 0.04 ± 0.02, WT+Ros responsive = 0.96 ± 0.02 (n=10). (E) Histogram of the fraction of silent boutons measured by GCaMP3 and normalized to fraction of WT silent boutons in WT, CNAαKD, CDK5KD, WT with/without CSA or Roscovitine, WT with Cono or Aga with/without Roscovitine.: WT = 1.0 ± 0.18 (n=25), CNAαKD = 3.00 ± 0.24 (n=26), CDK5KD = 0.36 ± 0.09 (n= 12), WT+CSA = 2.70 ± 0.54 (n= 9), WT+Ros = 0.40 ± 0.09 (n= 16), WT+Cono = 3.31 ± 0.66, WT+Cono +Ros = 2.85 ± 0.60 (n= 7), WT+Aga = 1.87 ± 0.16, WT+Aga+Ros = 0.76 ± 0.40 (n= 8).
Figure Legend Snippet: CNAα and CDK5 gate silent presynaptic boutons by controlling Ca 2+ influx (A) Representative single-AP GCaMP3 difference image (ΔF 1AP ) in WT neurons in the absence or presence of CSA (Top) or Ros (Bottom). Ionomycin treatment (right) reveals all presynaptic boutons. White circles indicate boutons that were silenced after treatment with CSA (Top) or unsilenced after treatment of Ros (Bottom). Scale bar represents 5 μm. (B) Corresponding traces of silenced (white circle in A top) and unsilenced boutons (white circle in A bottom) with or without treatment of CSA or Ros. (C) Histogram of the fraction of silent and responsive boutons measured by GCaMP3 in WT with or without CSA or Roscovitine: WT silent = 0.17 ± 0.04, WT responsive = 0.83 ± 0.04 (n= 25), WT+CSA silent = 0.46 ± 0.09, WT+CSA responsive = 0.54 ± 0.09 (n= 9), WT+Ros silent = 0.07 ± 0.02, WT+Ros responsive = 0.93 ± 0.02 (n= 16). (D) Histogram of the proportion of silent and responsive boutons measured by MgGreen in WT with or without CSA or Ros. The division of silent or responsive boutons was based on a signal to noise ratio of 1 (peak versus SD of baseline) for the single-AP response: WT silent = 0.14 ± 0.03, WT responsive = 0.86 ± 0.03 (n=19), WT+CSA silent = 0.36 ± 0.11, WT+CSA responsive = 0.64 ± 0.11 (n=7), WT+Ros silent = 0.04 ± 0.02, WT+Ros responsive = 0.96 ± 0.02 (n=10). (E) Histogram of the fraction of silent boutons measured by GCaMP3 and normalized to fraction of WT silent boutons in WT, CNAαKD, CDK5KD, WT with/without CSA or Roscovitine, WT with Cono or Aga with/without Roscovitine.: WT = 1.0 ± 0.18 (n=25), CNAαKD = 3.00 ± 0.24 (n=26), CDK5KD = 0.36 ± 0.09 (n= 12), WT+CSA = 2.70 ± 0.54 (n= 9), WT+Ros = 0.40 ± 0.09 (n= 16), WT+Cono = 3.31 ± 0.66, WT+Cono +Ros = 2.85 ± 0.60 (n= 7), WT+Aga = 1.87 ± 0.16, WT+Aga+Ros = 0.76 ± 0.40 (n= 8).

Techniques Used:

3) Product Images from "Super-resolution imaging of live sperm reveals dynamic changes of the actin cytoskeleton during acrosomal exocytosis"

Article Title: Super-resolution imaging of live sperm reveals dynamic changes of the actin cytoskeleton during acrosomal exocytosis

Journal: Journal of Cell Science

doi: 10.1242/jcs.218958

Specific F-actin structures in the sperm head did not change as a consequence of the AR initiation. Sperm were loaded with SiR-actin (green) and FM4-64 (magenta) and attached to concanavalin A-coated slides for imaging using TIRF microscopy. Following image acquisition, the ROIs indicated in A and D were analyzed. Representative image sequences of sperm stimulated with ionomycin (addition indicated by black arrow) that initially possessed the F-actin structure corresponding to the perforatorium (B), ventral (D; i) or neck (D; ii) region. The corresponding fluorescence traces (C and F) of the indicated ROIs (for SiR-actin) and the whole sperm head (for FM4-64) (A and D) are shown on the right. Analysis of these traces demonstrates that the depolymerization did not occur before or after the initiation of the AR (C and F) ( n =6. 62 cells analyzed).
Figure Legend Snippet: Specific F-actin structures in the sperm head did not change as a consequence of the AR initiation. Sperm were loaded with SiR-actin (green) and FM4-64 (magenta) and attached to concanavalin A-coated slides for imaging using TIRF microscopy. Following image acquisition, the ROIs indicated in A and D were analyzed. Representative image sequences of sperm stimulated with ionomycin (addition indicated by black arrow) that initially possessed the F-actin structure corresponding to the perforatorium (B), ventral (D; i) or neck (D; ii) region. The corresponding fluorescence traces (C and F) of the indicated ROIs (for SiR-actin) and the whole sperm head (for FM4-64) (A and D) are shown on the right. Analysis of these traces demonstrates that the depolymerization did not occur before or after the initiation of the AR (C and F) ( n =6. 62 cells analyzed).

Techniques Used: Imaging, Microscopy, Fluorescence

Specific dynamic changes in the actin cytoskeleton after initiation of acrosomal exocytosis. (A) Sperm labeled with SiR-actin (green) and FM4-64 (magenta). The ROIs used for the analysis, corresponding to the upper acrosome region (for SiR-actin) and the whole sperm head (for FM4-64), are depicted. (B) Representative sequence of images of a sperm loaded with SiR-actin and FM4-64 that displayed a decrease in SiR-actin fluorescence in the upper acrosome region after stimulation with 10 µM ionomycin (addition indicated by black arrow). ″ represents time in seconds. (C) The resulting fluorescence traces revealed that the loss of F-actin occurred after the initiation of the AR as judged by the increase in FM4-64 fluorescence ( n .
Figure Legend Snippet: Specific dynamic changes in the actin cytoskeleton after initiation of acrosomal exocytosis. (A) Sperm labeled with SiR-actin (green) and FM4-64 (magenta). The ROIs used for the analysis, corresponding to the upper acrosome region (for SiR-actin) and the whole sperm head (for FM4-64), are depicted. (B) Representative sequence of images of a sperm loaded with SiR-actin and FM4-64 that displayed a decrease in SiR-actin fluorescence in the upper acrosome region after stimulation with 10 µM ionomycin (addition indicated by black arrow). ″ represents time in seconds. (C) The resulting fluorescence traces revealed that the loss of F-actin occurred after the initiation of the AR as judged by the increase in FM4-64 fluorescence ( n .

Techniques Used: Labeling, Sequencing, Fluorescence

4) Product Images from "A Specific Transitory Increase in Intracellular Calcium Induced by Progesterone Promotes Acrosomal Exocytosis in Mouse Sperm 1"

Article Title: A Specific Transitory Increase in Intracellular Calcium Induced by Progesterone Promotes Acrosomal Exocytosis in Mouse Sperm 1

Journal: Biology of Reproduction

doi: 10.1095/biolreprod.115.136085

Directionality of the [Ca 2+ ]i increase wave propagation in response to progesterone in mouse sperm immobilized with concanavalin A. A ) Representative diagram of the different sperm areas evaluated in sperm attached to concanavalin A-coated slides. PPP, posterior principal piece; PPP II , posterior principal piece II; PPP I , posterior principal piece I; APP, anterior principal piece; CD, cytoplasmic droplet; PMP, posterior midpiece; AMP, anterior midpiece; H, head. B and E ) Representative images of the time course of Fluo-4 AM-labeled sperm after addition of 100 μM progesterone or 10 μM ionomycin as a control, respectively. C and F ) Time course of the fluorescence changes in each of the areas of the sperm shown in B and E , respectively. D and G ) Different time delays of the analyzed areas of the [Ca 2+ ]i increase propagation in sperm stimulated with progesterone or ionomycin, respectively (n = 8). Statistically significant differences at * P
Figure Legend Snippet: Directionality of the [Ca 2+ ]i increase wave propagation in response to progesterone in mouse sperm immobilized with concanavalin A. A ) Representative diagram of the different sperm areas evaluated in sperm attached to concanavalin A-coated slides. PPP, posterior principal piece; PPP II , posterior principal piece II; PPP I , posterior principal piece I; APP, anterior principal piece; CD, cytoplasmic droplet; PMP, posterior midpiece; AMP, anterior midpiece; H, head. B and E ) Representative images of the time course of Fluo-4 AM-labeled sperm after addition of 100 μM progesterone or 10 μM ionomycin as a control, respectively. C and F ) Time course of the fluorescence changes in each of the areas of the sperm shown in B and E , respectively. D and G ) Different time delays of the analyzed areas of the [Ca 2+ ]i increase propagation in sperm stimulated with progesterone or ionomycin, respectively (n = 8). Statistically significant differences at * P

Techniques Used: Labeling, Fluorescence

[Ca 2+ ]i and AR alternate measurements in mouse sperm. A and B ) Fluo-4 AM and FM4-64 fluorescence from sperm exposed to progesterone (PROG) or ionomycin (IONO), respectively. The arrow in the panel corresponding to 75 sec indicates a sperm that increased its [Ca 2+ ]i and underwent AR (arrow in panel corresponding to 350 sec). C ) Representative alternate Fluo-4 AM and FM4-64 fluorescence images of a sperm undergoing a progesterone response. C ) Fluo-4 AM and FM4-64 fluorescence changes observed in the sperm displayed in D . E ) Alternate Fluo-4 AM and FM4-64 fluorescence images of a representative sperm responding to ionomycin. F ) Fluo-4 AM and FM4-64 fluorescence changes observed during the recordings of the sperm shown in E . G ) Percentage of AR stimulated by progesterone or ionomycin at different times (AR time was established as the time that FM4-64 fluorescence significantly increased); n = 8 mice, 21 reacted cells and n = 4 mice, 43 reacted cells for progesterone and ionomycin experiments, respectively. Statistically significant differences between the progesterone and ionomycin response at * P
Figure Legend Snippet: [Ca 2+ ]i and AR alternate measurements in mouse sperm. A and B ) Fluo-4 AM and FM4-64 fluorescence from sperm exposed to progesterone (PROG) or ionomycin (IONO), respectively. The arrow in the panel corresponding to 75 sec indicates a sperm that increased its [Ca 2+ ]i and underwent AR (arrow in panel corresponding to 350 sec). C ) Representative alternate Fluo-4 AM and FM4-64 fluorescence images of a sperm undergoing a progesterone response. C ) Fluo-4 AM and FM4-64 fluorescence changes observed in the sperm displayed in D . E ) Alternate Fluo-4 AM and FM4-64 fluorescence images of a representative sperm responding to ionomycin. F ) Fluo-4 AM and FM4-64 fluorescence changes observed during the recordings of the sperm shown in E . G ) Percentage of AR stimulated by progesterone or ionomycin at different times (AR time was established as the time that FM4-64 fluorescence significantly increased); n = 8 mice, 21 reacted cells and n = 4 mice, 43 reacted cells for progesterone and ionomycin experiments, respectively. Statistically significant differences between the progesterone and ionomycin response at * P

Techniques Used: Fluorescence, Size-exclusion Chromatography, Mouse Assay

Validation of the plasma membrane probe FM4-64 as an AR indicator and its simultaneous observation with the calcium indicator Fluo-4 AM. A ) Representative fluorescence images of the time course of an FM4-64-labeled EGFP-sperm that does not undergo AR after addition of 10 μM ionomycin (IONO). B ) EGFP and FM4-64 fluorescence intensities (in arbitrary units) of the sperm shown in A . C ) Representative fluorescence images of the time course of FM4-64-labeled EGFP-sperm undergoing AR after addition of 10 μM ionomycin. D ) EGFP and FM4-64 fluorescence intensities (in arbitrary units) of the sperm shown in C . E ) Fluorescence images of FM4-64- and Fluo-4 AM-labeled sperm.
Figure Legend Snippet: Validation of the plasma membrane probe FM4-64 as an AR indicator and its simultaneous observation with the calcium indicator Fluo-4 AM. A ) Representative fluorescence images of the time course of an FM4-64-labeled EGFP-sperm that does not undergo AR after addition of 10 μM ionomycin (IONO). B ) EGFP and FM4-64 fluorescence intensities (in arbitrary units) of the sperm shown in A . C ) Representative fluorescence images of the time course of FM4-64-labeled EGFP-sperm undergoing AR after addition of 10 μM ionomycin. D ) EGFP and FM4-64 fluorescence intensities (in arbitrary units) of the sperm shown in C . E ) Fluorescence images of FM4-64- and Fluo-4 AM-labeled sperm.

Techniques Used: Fluorescence, Labeling

Mouse sperm displayed different patterns of [Ca 2+ ]i increase in response to progesterone. A ) Graphics representing the different patterns of [Ca 2+ ]i increase observed in the sperm head as a result of the addition of 100 μM progesterone (PROG). According to the calcium response, the patterns of increase were classified as sustained, transitory, oscillatory, late transitory, and gradual increase. The bar below the traces represents the time scale = 1 min. B ) Representative time lapse images of Fluo-4 AM-loaded sperm following the addition of 100 μM progesterone and 10 μM ionomycin (IONO). C ) Percentage of sperm displaying each of the patterns observed. These results were obtained by analyzing 184 sperm from 10 mice. D ) Comparison of the calcium increase in response to progesterone normalized by the ionomycin response. These results were obtained by analyzing 184 sperm from 10 mice; statistically significant differences at * P
Figure Legend Snippet: Mouse sperm displayed different patterns of [Ca 2+ ]i increase in response to progesterone. A ) Graphics representing the different patterns of [Ca 2+ ]i increase observed in the sperm head as a result of the addition of 100 μM progesterone (PROG). According to the calcium response, the patterns of increase were classified as sustained, transitory, oscillatory, late transitory, and gradual increase. The bar below the traces represents the time scale = 1 min. B ) Representative time lapse images of Fluo-4 AM-loaded sperm following the addition of 100 μM progesterone and 10 μM ionomycin (IONO). C ) Percentage of sperm displaying each of the patterns observed. These results were obtained by analyzing 184 sperm from 10 mice. D ) Comparison of the calcium increase in response to progesterone normalized by the ionomycin response. These results were obtained by analyzing 184 sperm from 10 mice; statistically significant differences at * P

Techniques Used: Mouse Assay

Progesterone promotes an [Ca 2+ ]i increase in Fluo-4 AM-loaded mice sperm. A ) Image sequence showing sperm exposed to 40 μM progesterone (PROG) and recorded for 30 min. Sperm displayed an [Ca 2+ ]i increase at different times during the experiment. At the end of the recording, ionomycin (IONO) was added as a viability control. B ) Fluorescence changes corresponding to sperm 1–3 of the field recorded in A . C ) Percentage of sperm displaying an [Ca 2+ ]i increase in response to vehicle, 40 μM, and 100 μM progesterone addition. Black arrows indicate the time where progesterone and ionomycin were applied. These results were obtained by analyzing 319 sperm from 25 mice; statistically significant differences at * P
Figure Legend Snippet: Progesterone promotes an [Ca 2+ ]i increase in Fluo-4 AM-loaded mice sperm. A ) Image sequence showing sperm exposed to 40 μM progesterone (PROG) and recorded for 30 min. Sperm displayed an [Ca 2+ ]i increase at different times during the experiment. At the end of the recording, ionomycin (IONO) was added as a viability control. B ) Fluorescence changes corresponding to sperm 1–3 of the field recorded in A . C ) Percentage of sperm displaying an [Ca 2+ ]i increase in response to vehicle, 40 μM, and 100 μM progesterone addition. Black arrows indicate the time where progesterone and ionomycin were applied. These results were obtained by analyzing 319 sperm from 25 mice; statistically significant differences at * P

Techniques Used: Mouse Assay, Sequencing, Fluorescence

5) Product Images from "Acrosome Reaction and Ca2+ Imaging in Single Human Spermatozoa: New Regulatory Roles of [Ca2+]i 1"

Article Title: Acrosome Reaction and Ca2+ Imaging in Single Human Spermatozoa: New Regulatory Roles of [Ca2+]i 1

Journal: Biology of Reproduction

doi: 10.1095/biolreprod.114.119768

AR and [Ca 2+ ]i imaging in single human spermatozoa exposed to ionomycin and progesterone. A ) Nonstimulated human spermatozoa labeled with FM4-64 (red) and Fluo-4 (green). FM4-64 labels the plasma membrane, and Fluo-4 is distributed mainly in the cytoplasm.
Figure Legend Snippet: AR and [Ca 2+ ]i imaging in single human spermatozoa exposed to ionomycin and progesterone. A ) Nonstimulated human spermatozoa labeled with FM4-64 (red) and Fluo-4 (green). FM4-64 labels the plasma membrane, and Fluo-4 is distributed mainly in the cytoplasm.

Techniques Used: Imaging, Labeling

Evidence that FM4-64 acrosome staining corresponds to the onset of the AR. A ) Concomitant imaging of FM4-64 and FITC-PSA during ionomycin application to human spermatozoa. Ionomycin applied at ∼80 sec (black arrowhead indicates its addition) increases
Figure Legend Snippet: Evidence that FM4-64 acrosome staining corresponds to the onset of the AR. A ) Concomitant imaging of FM4-64 and FITC-PSA during ionomycin application to human spermatozoa. Ionomycin applied at ∼80 sec (black arrowhead indicates its addition) increases

Techniques Used: Staining, Imaging, Size-exclusion Chromatography

Related Articles

other:

Article Title: A Specific Transitory Increase in Intracellular Calcium Induced by Progesterone Promotes Acrosomal Exocytosis in Mouse Sperm 1
Article Snippet: Bovine serum albumin, concanavalin A, and progesterone were purchased from Sigma-Aldrich Chemical Co. Ionomycin was from Alomone Labs.

Article Title: Super-resolution imaging of live sperm reveals dynamic changes of the actin cytoskeleton during acrosomal exocytosis
Article Snippet: Ionomycin was from Alomone Laboratories (Jerusalem, Israel).

Article Title: Acrosome Reaction and Ca2+ Imaging in Single Human Spermatozoa: New Regulatory Roles of [Ca2+]i 1
Article Snippet: Bovine serum albumin, Ham F-10 media, FITC-PSA, and progesterone were purchased from Sigma-Aldrich Chemical Co. Ionomycin was from Alomone Labs.

Mass Spectrometry:

Article Title: TRPV4-mediates oscillatory fluid shear mechanotransduction in mesenchymal stem cells in part via the primary cilium
Article Snippet: .. Exposure time was kept below 600 ms and was kept constant between control and treatment groups, allowing image acquisition every 1.29 s. Following each experiment, 10 µM ionomycin (Alomone Labs, Jerusalem, Israel, http://www.alomone.com ) was applied to the cells as a positive control for sensitivity of the indicator to calcium. .. Response was calculated as fold change fluorescence over baseline levels where baseline was taken as the 30 seconds prior to treatment (t = 0).

Positive Control:

Article Title: TRPV4-mediates oscillatory fluid shear mechanotransduction in mesenchymal stem cells in part via the primary cilium
Article Snippet: .. Exposure time was kept below 600 ms and was kept constant between control and treatment groups, allowing image acquisition every 1.29 s. Following each experiment, 10 µM ionomycin (Alomone Labs, Jerusalem, Israel, http://www.alomone.com ) was applied to the cells as a positive control for sensitivity of the indicator to calcium. .. Response was calculated as fold change fluorescence over baseline levels where baseline was taken as the 30 seconds prior to treatment (t = 0).

Similar Products

  • Logo
  • About
  • News
  • Press Release
  • Team
  • Advisors
  • Partners
  • Contact
  • Bioz Stars
  • Bioz vStars
  • 91
    Alomone Labs ionomycin
    Specific F-actin structures in the sperm head did not change as a consequence of the AR initiation. Sperm were loaded with SiR-actin (green) and FM4-64 (magenta) and attached to concanavalin A-coated slides for imaging using TIRF microscopy. Following image acquisition, the ROIs indicated in A and D were analyzed. Representative image sequences of sperm stimulated with <t>ionomycin</t> (addition indicated by black arrow) that initially possessed the F-actin structure corresponding to the perforatorium (B), ventral (D; i) or neck (D; ii) region. The corresponding fluorescence traces (C and F) of the indicated ROIs (for SiR-actin) and the whole sperm head (for FM4-64) (A and D) are shown on the right. Analysis of these traces demonstrates that the depolymerization did not occur before or after the initiation of the AR (C and F) ( n =6. 62 cells analyzed).
    Ionomycin, supplied by Alomone Labs, used in various techniques. Bioz Stars score: 91/100, based on 3 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/ionomycin/product/Alomone Labs
    Average 91 stars, based on 3 article reviews
    Price from $9.99 to $1999.99
    ionomycin - by Bioz Stars, 2020-09
    91/100 stars
      Buy from Supplier

    Image Search Results


    Specific F-actin structures in the sperm head did not change as a consequence of the AR initiation. Sperm were loaded with SiR-actin (green) and FM4-64 (magenta) and attached to concanavalin A-coated slides for imaging using TIRF microscopy. Following image acquisition, the ROIs indicated in A and D were analyzed. Representative image sequences of sperm stimulated with ionomycin (addition indicated by black arrow) that initially possessed the F-actin structure corresponding to the perforatorium (B), ventral (D; i) or neck (D; ii) region. The corresponding fluorescence traces (C and F) of the indicated ROIs (for SiR-actin) and the whole sperm head (for FM4-64) (A and D) are shown on the right. Analysis of these traces demonstrates that the depolymerization did not occur before or after the initiation of the AR (C and F) ( n =6. 62 cells analyzed).

    Journal: Journal of Cell Science

    Article Title: Super-resolution imaging of live sperm reveals dynamic changes of the actin cytoskeleton during acrosomal exocytosis

    doi: 10.1242/jcs.218958

    Figure Lengend Snippet: Specific F-actin structures in the sperm head did not change as a consequence of the AR initiation. Sperm were loaded with SiR-actin (green) and FM4-64 (magenta) and attached to concanavalin A-coated slides for imaging using TIRF microscopy. Following image acquisition, the ROIs indicated in A and D were analyzed. Representative image sequences of sperm stimulated with ionomycin (addition indicated by black arrow) that initially possessed the F-actin structure corresponding to the perforatorium (B), ventral (D; i) or neck (D; ii) region. The corresponding fluorescence traces (C and F) of the indicated ROIs (for SiR-actin) and the whole sperm head (for FM4-64) (A and D) are shown on the right. Analysis of these traces demonstrates that the depolymerization did not occur before or after the initiation of the AR (C and F) ( n =6. 62 cells analyzed).

    Article Snippet: Ionomycin was from Alomone Laboratories (Jerusalem, Israel).

    Techniques: Imaging, Microscopy, Fluorescence

    Specific dynamic changes in the actin cytoskeleton after initiation of acrosomal exocytosis. (A) Sperm labeled with SiR-actin (green) and FM4-64 (magenta). The ROIs used for the analysis, corresponding to the upper acrosome region (for SiR-actin) and the whole sperm head (for FM4-64), are depicted. (B) Representative sequence of images of a sperm loaded with SiR-actin and FM4-64 that displayed a decrease in SiR-actin fluorescence in the upper acrosome region after stimulation with 10†µM ionomycin (addition indicated by black arrow). ″ represents time in seconds. (C) The resulting fluorescence traces revealed that the loss of F-actin occurred after the initiation of the AR as judged by the increase in FM4-64 fluorescence ( n .

    Journal: Journal of Cell Science

    Article Title: Super-resolution imaging of live sperm reveals dynamic changes of the actin cytoskeleton during acrosomal exocytosis

    doi: 10.1242/jcs.218958

    Figure Lengend Snippet: Specific dynamic changes in the actin cytoskeleton after initiation of acrosomal exocytosis. (A) Sperm labeled with SiR-actin (green) and FM4-64 (magenta). The ROIs used for the analysis, corresponding to the upper acrosome region (for SiR-actin) and the whole sperm head (for FM4-64), are depicted. (B) Representative sequence of images of a sperm loaded with SiR-actin and FM4-64 that displayed a decrease in SiR-actin fluorescence in the upper acrosome region after stimulation with 10†µM ionomycin (addition indicated by black arrow). ″ represents time in seconds. (C) The resulting fluorescence traces revealed that the loss of F-actin occurred after the initiation of the AR as judged by the increase in FM4-64 fluorescence ( n .

    Article Snippet: Ionomycin was from Alomone Laboratories (Jerusalem, Israel).

    Techniques: Labeling, Sequencing, Fluorescence

    CNAα and CDK5 gate silent presynaptic boutons by controlling Ca 2+ influx (A) Representative single-AP GCaMP3 difference image (ΔF 1AP ) in WT neurons in the absence or presence of CSA (Top) or Ros (Bottom). Ionomycin treatment (right) reveals all presynaptic boutons. White circles indicate boutons that were silenced after treatment with CSA (Top) or unsilenced after treatment of Ros (Bottom). Scale bar represents 5 μm. (B) Corresponding traces of silenced (white circle in A top) and unsilenced boutons (white circle in A bottom) with or without treatment of CSA or Ros. (C) Histogram of the fraction of silent and responsive boutons measured by GCaMP3 in WT with or without CSA or Roscovitine: WT silent = 0.17 ± 0.04, WT responsive = 0.83 ± 0.04 (n= 25), WT+CSA silent = 0.46 ± 0.09, WT+CSA responsive = 0.54 ± 0.09 (n= 9), WT+Ros silent = 0.07 ± 0.02, WT+Ros responsive = 0.93 ± 0.02 (n= 16). (D) Histogram of the proportion of silent and responsive boutons measured by MgGreen in WT with or without CSA or Ros. The division of silent or responsive boutons was based on a signal to noise ratio of 1 (peak versus SD of baseline) for the single-AP response: WT silent = 0.14 ± 0.03, WT responsive = 0.86 ± 0.03 (n=19), WT+CSA silent = 0.36 ± 0.11, WT+CSA responsive = 0.64 ± 0.11 (n=7), WT+Ros silent = 0.04 ± 0.02, WT+Ros responsive = 0.96 ± 0.02 (n=10). (E) Histogram of the fraction of silent boutons measured by GCaMP3 and normalized to fraction of WT silent boutons in WT, CNAαKD, CDK5KD, WT with/without CSA or Roscovitine, WT with Cono or Aga with/without Roscovitine.: WT = 1.0 ± 0.18 (n=25), CNAαKD = 3.00 ± 0.24 (n=26), CDK5KD = 0.36 ± 0.09 (n= 12), WT+CSA = 2.70 ± 0.54 (n= 9), WT+Ros = 0.40 ± 0.09 (n= 16), WT+Cono = 3.31 ± 0.66, WT+Cono +Ros = 2.85 ± 0.60 (n= 7), WT+Aga = 1.87 ± 0.16, WT+Aga+Ros = 0.76 ± 0.40 (n= 8).

    Journal: The Journal of neuroscience : the official journal of the Society for Neuroscience

    Article Title: Balance of calcineurin A? and CDK5 activities sets release probability at nerve terminals

    doi: 10.1523/JNEUROSCI.4288-12.2013

    Figure Lengend Snippet: CNAα and CDK5 gate silent presynaptic boutons by controlling Ca 2+ influx (A) Representative single-AP GCaMP3 difference image (ΔF 1AP ) in WT neurons in the absence or presence of CSA (Top) or Ros (Bottom). Ionomycin treatment (right) reveals all presynaptic boutons. White circles indicate boutons that were silenced after treatment with CSA (Top) or unsilenced after treatment of Ros (Bottom). Scale bar represents 5 μm. (B) Corresponding traces of silenced (white circle in A top) and unsilenced boutons (white circle in A bottom) with or without treatment of CSA or Ros. (C) Histogram of the fraction of silent and responsive boutons measured by GCaMP3 in WT with or without CSA or Roscovitine: WT silent = 0.17 ± 0.04, WT responsive = 0.83 ± 0.04 (n= 25), WT+CSA silent = 0.46 ± 0.09, WT+CSA responsive = 0.54 ± 0.09 (n= 9), WT+Ros silent = 0.07 ± 0.02, WT+Ros responsive = 0.93 ± 0.02 (n= 16). (D) Histogram of the proportion of silent and responsive boutons measured by MgGreen in WT with or without CSA or Ros. The division of silent or responsive boutons was based on a signal to noise ratio of 1 (peak versus SD of baseline) for the single-AP response: WT silent = 0.14 ± 0.03, WT responsive = 0.86 ± 0.03 (n=19), WT+CSA silent = 0.36 ± 0.11, WT+CSA responsive = 0.64 ± 0.11 (n=7), WT+Ros silent = 0.04 ± 0.02, WT+Ros responsive = 0.96 ± 0.02 (n=10). (E) Histogram of the fraction of silent boutons measured by GCaMP3 and normalized to fraction of WT silent boutons in WT, CNAαKD, CDK5KD, WT with/without CSA or Roscovitine, WT with Cono or Aga with/without Roscovitine.: WT = 1.0 ± 0.18 (n=25), CNAαKD = 3.00 ± 0.24 (n=26), CDK5KD = 0.36 ± 0.09 (n= 12), WT+CSA = 2.70 ± 0.54 (n= 9), WT+Ros = 0.40 ± 0.09 (n= 16), WT+Cono = 3.31 ± 0.66, WT+Cono +Ros = 2.85 ± 0.60 (n= 7), WT+Aga = 1.87 ± 0.16, WT+Aga+Ros = 0.76 ± 0.40 (n= 8).

    Article Snippet: Unless otherwise noted, all chemicals were obtained from Sigma (St. Louis, MO) except for Ca2+ channel toxins, ω-AagtoxinIVA, ω-Conotoxin GVIA (Alomone Labs), Ionomycin (Alomone Labs), Ca2+ dyes, Fluo5F AM and Magnesium Green AM (invitrogen) and EGTA AM (invitrogen).

    Techniques:

    Directionality of the [Ca 2+ ]i increase wave propagation in response to progesterone in mouse sperm immobilized with concanavalin A. A ) Representative diagram of the different sperm areas evaluated in sperm attached to concanavalin A-coated slides. PPP, posterior principal piece; PPP II , posterior principal piece II; PPP I , posterior principal piece I; APP, anterior principal piece; CD, cytoplasmic droplet; PMP, posterior midpiece; AMP, anterior midpiece; H, head. B and E ) Representative images of the time course of Fluo-4 AM-labeled sperm after addition of 100 μM progesterone or 10 μM ionomycin as a control, respectively. C and F ) Time course of the fluorescence changes in each of the areas of the sperm shown in B and E , respectively. D and G ) Different time delays of the analyzed areas of the [Ca 2+ ]i increase propagation in sperm stimulated with progesterone or ionomycin, respectively (n = 8). Statistically significant differences at * P

    Journal: Biology of Reproduction

    Article Title: A Specific Transitory Increase in Intracellular Calcium Induced by Progesterone Promotes Acrosomal Exocytosis in Mouse Sperm 1

    doi: 10.1095/biolreprod.115.136085

    Figure Lengend Snippet: Directionality of the [Ca 2+ ]i increase wave propagation in response to progesterone in mouse sperm immobilized with concanavalin A. A ) Representative diagram of the different sperm areas evaluated in sperm attached to concanavalin A-coated slides. PPP, posterior principal piece; PPP II , posterior principal piece II; PPP I , posterior principal piece I; APP, anterior principal piece; CD, cytoplasmic droplet; PMP, posterior midpiece; AMP, anterior midpiece; H, head. B and E ) Representative images of the time course of Fluo-4 AM-labeled sperm after addition of 100 μM progesterone or 10 μM ionomycin as a control, respectively. C and F ) Time course of the fluorescence changes in each of the areas of the sperm shown in B and E , respectively. D and G ) Different time delays of the analyzed areas of the [Ca 2+ ]i increase propagation in sperm stimulated with progesterone or ionomycin, respectively (n = 8). Statistically significant differences at * P

    Article Snippet: Bovine serum albumin, concanavalin A, and progesterone were purchased from Sigma-Aldrich Chemical Co. Ionomycin was from Alomone Labs.

    Techniques: Labeling, Fluorescence

    [Ca 2+ ]i and AR alternate measurements in mouse sperm. A and B ) Fluo-4 AM and FM4-64 fluorescence from sperm exposed to progesterone (PROG) or ionomycin (IONO), respectively. The arrow in the panel corresponding to 75 sec indicates a sperm that increased its [Ca 2+ ]i and underwent AR (arrow in panel corresponding to 350 sec). C ) Representative alternate Fluo-4 AM and FM4-64 fluorescence images of a sperm undergoing a progesterone response. C ) Fluo-4 AM and FM4-64 fluorescence changes observed in the sperm displayed in D . E ) Alternate Fluo-4 AM and FM4-64 fluorescence images of a representative sperm responding to ionomycin. F ) Fluo-4 AM and FM4-64 fluorescence changes observed during the recordings of the sperm shown in E . G ) Percentage of AR stimulated by progesterone or ionomycin at different times (AR time was established as the time that FM4-64 fluorescence significantly increased); n = 8 mice, 21 reacted cells and n = 4 mice, 43 reacted cells for progesterone and ionomycin experiments, respectively. Statistically significant differences between the progesterone and ionomycin response at * P

    Journal: Biology of Reproduction

    Article Title: A Specific Transitory Increase in Intracellular Calcium Induced by Progesterone Promotes Acrosomal Exocytosis in Mouse Sperm 1

    doi: 10.1095/biolreprod.115.136085

    Figure Lengend Snippet: [Ca 2+ ]i and AR alternate measurements in mouse sperm. A and B ) Fluo-4 AM and FM4-64 fluorescence from sperm exposed to progesterone (PROG) or ionomycin (IONO), respectively. The arrow in the panel corresponding to 75 sec indicates a sperm that increased its [Ca 2+ ]i and underwent AR (arrow in panel corresponding to 350 sec). C ) Representative alternate Fluo-4 AM and FM4-64 fluorescence images of a sperm undergoing a progesterone response. C ) Fluo-4 AM and FM4-64 fluorescence changes observed in the sperm displayed in D . E ) Alternate Fluo-4 AM and FM4-64 fluorescence images of a representative sperm responding to ionomycin. F ) Fluo-4 AM and FM4-64 fluorescence changes observed during the recordings of the sperm shown in E . G ) Percentage of AR stimulated by progesterone or ionomycin at different times (AR time was established as the time that FM4-64 fluorescence significantly increased); n = 8 mice, 21 reacted cells and n = 4 mice, 43 reacted cells for progesterone and ionomycin experiments, respectively. Statistically significant differences between the progesterone and ionomycin response at * P

    Article Snippet: Bovine serum albumin, concanavalin A, and progesterone were purchased from Sigma-Aldrich Chemical Co. Ionomycin was from Alomone Labs.

    Techniques: Fluorescence, Size-exclusion Chromatography, Mouse Assay

    Validation of the plasma membrane probe FM4-64 as an AR indicator and its simultaneous observation with the calcium indicator Fluo-4 AM. A ) Representative fluorescence images of the time course of an FM4-64-labeled EGFP-sperm that does not undergo AR after addition of 10 μM ionomycin (IONO). B ) EGFP and FM4-64 fluorescence intensities (in arbitrary units) of the sperm shown in A . C ) Representative fluorescence images of the time course of FM4-64-labeled EGFP-sperm undergoing AR after addition of 10 μM ionomycin. D ) EGFP and FM4-64 fluorescence intensities (in arbitrary units) of the sperm shown in C . E ) Fluorescence images of FM4-64- and Fluo-4 AM-labeled sperm.

    Journal: Biology of Reproduction

    Article Title: A Specific Transitory Increase in Intracellular Calcium Induced by Progesterone Promotes Acrosomal Exocytosis in Mouse Sperm 1

    doi: 10.1095/biolreprod.115.136085

    Figure Lengend Snippet: Validation of the plasma membrane probe FM4-64 as an AR indicator and its simultaneous observation with the calcium indicator Fluo-4 AM. A ) Representative fluorescence images of the time course of an FM4-64-labeled EGFP-sperm that does not undergo AR after addition of 10 μM ionomycin (IONO). B ) EGFP and FM4-64 fluorescence intensities (in arbitrary units) of the sperm shown in A . C ) Representative fluorescence images of the time course of FM4-64-labeled EGFP-sperm undergoing AR after addition of 10 μM ionomycin. D ) EGFP and FM4-64 fluorescence intensities (in arbitrary units) of the sperm shown in C . E ) Fluorescence images of FM4-64- and Fluo-4 AM-labeled sperm.

    Article Snippet: Bovine serum albumin, concanavalin A, and progesterone were purchased from Sigma-Aldrich Chemical Co. Ionomycin was from Alomone Labs.

    Techniques: Fluorescence, Labeling

    Mouse sperm displayed different patterns of [Ca 2+ ]i increase in response to progesterone. A ) Graphics representing the different patterns of [Ca 2+ ]i increase observed in the sperm head as a result of the addition of 100 μM progesterone (PROG). According to the calcium response, the patterns of increase were classified as sustained, transitory, oscillatory, late transitory, and gradual increase. The bar below the traces represents the time scale = 1 min. B ) Representative time lapse images of Fluo-4 AM-loaded sperm following the addition of 100 μM progesterone and 10 μM ionomycin (IONO). C ) Percentage of sperm displaying each of the patterns observed. These results were obtained by analyzing 184 sperm from 10 mice. D ) Comparison of the calcium increase in response to progesterone normalized by the ionomycin response. These results were obtained by analyzing 184 sperm from 10 mice; statistically significant differences at * P

    Journal: Biology of Reproduction

    Article Title: A Specific Transitory Increase in Intracellular Calcium Induced by Progesterone Promotes Acrosomal Exocytosis in Mouse Sperm 1

    doi: 10.1095/biolreprod.115.136085

    Figure Lengend Snippet: Mouse sperm displayed different patterns of [Ca 2+ ]i increase in response to progesterone. A ) Graphics representing the different patterns of [Ca 2+ ]i increase observed in the sperm head as a result of the addition of 100 μM progesterone (PROG). According to the calcium response, the patterns of increase were classified as sustained, transitory, oscillatory, late transitory, and gradual increase. The bar below the traces represents the time scale = 1 min. B ) Representative time lapse images of Fluo-4 AM-loaded sperm following the addition of 100 μM progesterone and 10 μM ionomycin (IONO). C ) Percentage of sperm displaying each of the patterns observed. These results were obtained by analyzing 184 sperm from 10 mice. D ) Comparison of the calcium increase in response to progesterone normalized by the ionomycin response. These results were obtained by analyzing 184 sperm from 10 mice; statistically significant differences at * P

    Article Snippet: Bovine serum albumin, concanavalin A, and progesterone were purchased from Sigma-Aldrich Chemical Co. Ionomycin was from Alomone Labs.

    Techniques: Mouse Assay

    Progesterone promotes an [Ca 2+ ]i increase in Fluo-4 AM-loaded mice sperm. A ) Image sequence showing sperm exposed to 40 μM progesterone (PROG) and recorded for 30 min. Sperm displayed an [Ca 2+ ]i increase at different times during the experiment. At the end of the recording, ionomycin (IONO) was added as a viability control. B ) Fluorescence changes corresponding to sperm 1–3 of the field recorded in A . C ) Percentage of sperm displaying an [Ca 2+ ]i increase in response to vehicle, 40 μM, and 100 μM progesterone addition. Black arrows indicate the time where progesterone and ionomycin were applied. These results were obtained by analyzing 319 sperm from 25 mice; statistically significant differences at * P

    Journal: Biology of Reproduction

    Article Title: A Specific Transitory Increase in Intracellular Calcium Induced by Progesterone Promotes Acrosomal Exocytosis in Mouse Sperm 1

    doi: 10.1095/biolreprod.115.136085

    Figure Lengend Snippet: Progesterone promotes an [Ca 2+ ]i increase in Fluo-4 AM-loaded mice sperm. A ) Image sequence showing sperm exposed to 40 μM progesterone (PROG) and recorded for 30 min. Sperm displayed an [Ca 2+ ]i increase at different times during the experiment. At the end of the recording, ionomycin (IONO) was added as a viability control. B ) Fluorescence changes corresponding to sperm 1–3 of the field recorded in A . C ) Percentage of sperm displaying an [Ca 2+ ]i increase in response to vehicle, 40 μM, and 100 μM progesterone addition. Black arrows indicate the time where progesterone and ionomycin were applied. These results were obtained by analyzing 319 sperm from 25 mice; statistically significant differences at * P

    Article Snippet: Bovine serum albumin, concanavalin A, and progesterone were purchased from Sigma-Aldrich Chemical Co. Ionomycin was from Alomone Labs.

    Techniques: Mouse Assay, Sequencing, Fluorescence