Structured Review

Millipore cacl2
Expanded ESI-FTICR mass spectra of CaM with ( A ) RS20, ( B ) A13L, ( C ) V11L, ( D ) V11F, and ( E ) R16L. The spectra were measured in 5 mM ammonium acetate buffer, pH 5.9, in the absence of <t>CaCl2</t> . The spectra show peaks originated from the CaM–peptide complexes at the 8+ charge-state and CaM at the 7+ charge-state in their actual intensities relative to each other.
Cacl2, supplied by Millipore, used in various techniques. Bioz Stars score: 78/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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Images

1) Product Images from "Relative Affinity Constants by Electrospray Ionization and Fourier Transform Ion Cyclotron Resonance Mass Spectrometry: Calmodulin Binding to Peptide Analogs of Myosin Light Chain Kinase"

Article Title: Relative Affinity Constants by Electrospray Ionization and Fourier Transform Ion Cyclotron Resonance Mass Spectrometry: Calmodulin Binding to Peptide Analogs of Myosin Light Chain Kinase

Journal:

doi:

Expanded ESI-FTICR mass spectra of CaM with ( A ) RS20, ( B ) A13L, ( C ) V11L, ( D ) V11F, and ( E ) R16L. The spectra were measured in 5 mM ammonium acetate buffer, pH 5.9, in the absence of CaCl2 . The spectra show peaks originated from the CaM–peptide complexes at the 8+ charge-state and CaM at the 7+ charge-state in their actual intensities relative to each other.
Figure Legend Snippet: Expanded ESI-FTICR mass spectra of CaM with ( A ) RS20, ( B ) A13L, ( C ) V11L, ( D ) V11F, and ( E ) R16L. The spectra were measured in 5 mM ammonium acetate buffer, pH 5.9, in the absence of CaCl2 . The spectra show peaks originated from the CaM–peptide complexes at the 8+ charge-state and CaM at the 7+ charge-state in their actual intensities relative to each other.

Techniques Used: Chick Chorioallantoic Membrane Assay

2) Product Images from "Adipose Stem Cell Microbeads as Production Sources for Chondrogenic Growth Factors"

Article Title: Adipose Stem Cell Microbeads as Production Sources for Chondrogenic Growth Factors

Journal: Journal of Stem Cells & Regenerative Medicine

doi:

Effect of Divalent Crosslinks in ASC Microbead on Growth Factor mRNA Levels and Production. (A) mRNA levels and (B) growth factor secretion/DNA content on day 7 from ASC microbeads crosslinked in CaCl2 Ca++) or BaCl2 (Ba++), cultured in growth medium and normalized to ASCs on tissue culture polystyrene (TCPS). (C) Growth factor retained within microbeads normalized to DNA content (n = 6 samples, mean±SE, *p
Figure Legend Snippet: Effect of Divalent Crosslinks in ASC Microbead on Growth Factor mRNA Levels and Production. (A) mRNA levels and (B) growth factor secretion/DNA content on day 7 from ASC microbeads crosslinked in CaCl2 Ca++) or BaCl2 (Ba++), cultured in growth medium and normalized to ASCs on tissue culture polystyrene (TCPS). (C) Growth factor retained within microbeads normalized to DNA content (n = 6 samples, mean±SE, *p

Techniques Used: Cell Culture

3) Product Images from "C - Reactive Protein Induced Rearrangement of Phosphatidylcholine on Nanoparticle Mimics of Lipoprotein Particles"

Article Title: C - Reactive Protein Induced Rearrangement of Phosphatidylcholine on Nanoparticle Mimics of Lipoprotein Particles

Journal:

doi: 10.1021/jp911617q

a) UV-vis spectra of Au-SO-PC-HT nanoparticles in 10 mM HEPES at pH 6.5 with 0.1% NaN3 and with increasing concentrations of CaCl2 in the presence of 45.5 nM CRP, b) plot of LSPR red shift with increasing concentration of CaCl2 [25 - 252 μ M] in the presence of 45.5 nM CRP, and c) UV-vis spectra of Au-SO-PC-HT with an increasing concentration of CaCl2 [0 – 800 μ M] in the absence of CRP.
Figure Legend Snippet: a) UV-vis spectra of Au-SO-PC-HT nanoparticles in 10 mM HEPES at pH 6.5 with 0.1% NaN3 and with increasing concentrations of CaCl2 in the presence of 45.5 nM CRP, b) plot of LSPR red shift with increasing concentration of CaCl2 [25 - 252 μ M] in the presence of 45.5 nM CRP, and c) UV-vis spectra of Au-SO-PC-HT with an increasing concentration of CaCl2 [0 – 800 μ M] in the absence of CRP.

Techniques Used: Concentration Assay

UV-vis spectra of i) 10 nm bare Au nanoparticles in water, ii) Au-SO-PC-HT nanoparticles in 10 mM HEPES at pH 6.5 with 0.1% NaN3 in 227 μ M CaCl2 , and iii) Au-SO-PC-HT nanoparticles in the presence of 45.5 nM CRP and 252 μ M CaCl2 .
Figure Legend Snippet: UV-vis spectra of i) 10 nm bare Au nanoparticles in water, ii) Au-SO-PC-HT nanoparticles in 10 mM HEPES at pH 6.5 with 0.1% NaN3 in 227 μ M CaCl2 , and iii) Au-SO-PC-HT nanoparticles in the presence of 45.5 nM CRP and 252 μ M CaCl2 .

Techniques Used:

4) Product Images from "Coaxial Extrusion Bioprinting of 3D Microfibrous Constructs with Cell-Favorable Gelatin Methacryloyl Microenvironments"

Article Title: Coaxial Extrusion Bioprinting of 3D Microfibrous Constructs with Cell-Favorable Gelatin Methacryloyl Microenvironments

Journal: Biofabrication

doi: 10.1088/1758-5090/aa9d44

Bioprinting performance of alginate bioinks. A) Printability map showing the effect of feeding rates of the CaCl2 solution and the alginate bioink. B-G) Photographs showing bioprinted hollow constructs with curved and straight channels, the feeding rates of the CaCl2 solution and the alginate bioink (equal rates) were (B, C) 900 μL min-1 , (D, E) 700 μL min-1 , and (F, G) 400 μL min-1 . H-M) Channel diameter of the hollow constructs as a function of (H) feeding rates of the CaCl2 solution and the alginate bioink (equal rates), (I) feeding rate of the CaCl2 solution, (J) feeding rate of the alginate, (K) concentration of the alginate bioink, (L) concentration of the CaCl2 solution, and (M) nozzle moving speed. Unless otherwise noted, the concentration of the CaCl2 solution and the alginate bioink, the feeding rates of the CaCl2 solution and the alginate bioink (equal rates), and the nozzle moving speed were kept constant at 6.0% and 1.0%, 500 μL min-1 and 500 mm min-1 , respectively. The core-sheath nozzle was made using 23G/28G needles.
Figure Legend Snippet: Bioprinting performance of alginate bioinks. A) Printability map showing the effect of feeding rates of the CaCl2 solution and the alginate bioink. B-G) Photographs showing bioprinted hollow constructs with curved and straight channels, the feeding rates of the CaCl2 solution and the alginate bioink (equal rates) were (B, C) 900 μL min-1 , (D, E) 700 μL min-1 , and (F, G) 400 μL min-1 . H-M) Channel diameter of the hollow constructs as a function of (H) feeding rates of the CaCl2 solution and the alginate bioink (equal rates), (I) feeding rate of the CaCl2 solution, (J) feeding rate of the alginate, (K) concentration of the alginate bioink, (L) concentration of the CaCl2 solution, and (M) nozzle moving speed. Unless otherwise noted, the concentration of the CaCl2 solution and the alginate bioink, the feeding rates of the CaCl2 solution and the alginate bioink (equal rates), and the nozzle moving speed were kept constant at 6.0% and 1.0%, 500 μL min-1 and 500 mm min-1 , respectively. The core-sheath nozzle was made using 23G/28G needles.

Techniques Used: Construct, Concentration Assay

5) Product Images from "Adipose Stem Cell Microbeads as Production Sources for Chondrogenic Growth Factors"

Article Title: Adipose Stem Cell Microbeads as Production Sources for Chondrogenic Growth Factors

Journal: Journal of Stem Cells & Regenerative Medicine

doi:

Effect of Divalent Crosslinks in ASC Microbead on Growth Factor mRNA Levels and Production. (A) mRNA levels and (B) growth factor secretion/DNA content on day 7 from ASC microbeads crosslinked in CaCl2 Ca++) or BaCl2 (Ba++), cultured in growth medium and normalized to ASCs on tissue culture polystyrene (TCPS). (C) Growth factor retained within microbeads normalized to DNA content (n = 6 samples, mean±SE, *p
Figure Legend Snippet: Effect of Divalent Crosslinks in ASC Microbead on Growth Factor mRNA Levels and Production. (A) mRNA levels and (B) growth factor secretion/DNA content on day 7 from ASC microbeads crosslinked in CaCl2 Ca++) or BaCl2 (Ba++), cultured in growth medium and normalized to ASCs on tissue culture polystyrene (TCPS). (C) Growth factor retained within microbeads normalized to DNA content (n = 6 samples, mean±SE, *p

Techniques Used: Cell Culture

6) Product Images from "DNA-Endonuclease Complex Dynamics by Simultaneous FRET and Fluorophore Intensity in Evanescent Field"

Article Title: DNA-Endonuclease Complex Dynamics by Simultaneous FRET and Fluorophore Intensity in Evanescent Field

Journal:

doi: 10.1016/j.bpj.2017.01.017

( A ) Ecl18kI tetramer-DNA structure (PBD entry 2GB7) with simulated AV clouds of Cy3 ( green ) and Cy5 ( red ) for “antiparallel” ( top ) and “parallel” ( bottom ) loop conformations. The indicated distances are those between the centers of the AV clouds. DNA extensions with dotted lines denote its topology in the two-loop conformations. ( B ) Experimental scheme of DNA fragment surface-immobilization and excitation through TIR with ALEX is shown; different signals corresponding to various DNA conformations are illustrated. ( C ) Representative single-molecule fluorescence intensity and EFRET time traces in the presence of 4 nM of Ecl18kI and 10 mM of CaCl2 are shown.
Figure Legend Snippet: ( A ) Ecl18kI tetramer-DNA structure (PBD entry 2GB7) with simulated AV clouds of Cy3 ( green ) and Cy5 ( red ) for “antiparallel” ( top ) and “parallel” ( bottom ) loop conformations. The indicated distances are those between the centers of the AV clouds. DNA extensions with dotted lines denote its topology in the two-loop conformations. ( B ) Experimental scheme of DNA fragment surface-immobilization and excitation through TIR with ALEX is shown; different signals corresponding to various DNA conformations are illustrated. ( C ) Representative single-molecule fluorescence intensity and EFRET time traces in the presence of 4 nM of Ecl18kI and 10 mM of CaCl2 are shown.

Techniques Used: Fluorescence

7) Product Images from "Sustained-release synthetic biomarkers for monitoring thrombosis and inflammation using point-of-care compatible readouts"

Article Title: Sustained-release synthetic biomarkers for monitoring thrombosis and inflammation using point-of-care compatible readouts

Journal:

doi: 10.1002/adfm.201505142

In vitro characterizations of PEG-peptide substrates ( a ) Dynamic light scattering measurements of PEG backbones. ( b ) PEG-T1Q (thrombin substrate) was exposed to recombinant proteases involved in the clotting cascade. Representative dequenching measurements after protease addition. ( c ) Sensors were responsive to the natural process of clotting by adding CaCl2 to activate clotting in human serum. Addition of the thrombin inhibitor, Argatroban, mitigated the cleavage of the substrates. ( d ) PEG-M1Q (MMP9 substrate) was exposed to MMPs. MMP9 cleavage of the substrate was blocked by the MMP inhibitor Marimastat.
Figure Legend Snippet: In vitro characterizations of PEG-peptide substrates ( a ) Dynamic light scattering measurements of PEG backbones. ( b ) PEG-T1Q (thrombin substrate) was exposed to recombinant proteases involved in the clotting cascade. Representative dequenching measurements after protease addition. ( c ) Sensors were responsive to the natural process of clotting by adding CaCl2 to activate clotting in human serum. Addition of the thrombin inhibitor, Argatroban, mitigated the cleavage of the substrates. ( d ) PEG-M1Q (MMP9 substrate) was exposed to MMPs. MMP9 cleavage of the substrate was blocked by the MMP inhibitor Marimastat.

Techniques Used: In Vitro, Recombinant, Coagulation

8) Product Images from "EGFR and ADAMs Cooperate to Regulate Shedding and Endocytic Trafficking of the Desmosomal Cadherin Desmoglein 2"

Article Title: EGFR and ADAMs Cooperate to Regulate Shedding and Endocytic Trafficking of the Desmosomal Cadherin Desmoglein 2

Journal:

doi: 10.1091/mbc.E08-04-0356

MMP inhibition promotes Dsg2 accumulation at cell–cell borders and strengthens intercellular adhesion. (A) SCC68 cells were treated overnight with DMSO, TAPI-0 (TAPI), or PKI166 (PKI) in medium containing 0.25 mM CaCl2 . Indirect immunofluorescence was used to determine the cellular localization of desmoglein 2 (Dsg2) and E-cadherin (E-cad). Bar, 20 μm. (B) Staining along the border was quantified as follows: one-third border occupied (black), two-thirds (gray), or entire (white) border. TAPI- and PKI-treated cells showed increased intercellular Dsg2, but not E-cad, localization. (C) Confluent monolayers of SCC68 cells were cultured overnight in the presence of DMSO, GM6001 (GM), or PKI in 0.09, 0.50, or 1.0 mM CaCl2 . Intercellular adhesion was measured by counting the number of fragments generated from dispase-released monolayers subjected to a defined amount of mechanical stress. At 0.50 mM and 1.0 mM calcium, significantly fewer fragments were produced in the TAPI- and PKI-treated monolayers (p < 0.05).
Figure Legend Snippet: MMP inhibition promotes Dsg2 accumulation at cell–cell borders and strengthens intercellular adhesion. (A) SCC68 cells were treated overnight with DMSO, TAPI-0 (TAPI), or PKI166 (PKI) in medium containing 0.25 mM CaCl2 . Indirect immunofluorescence was used to determine the cellular localization of desmoglein 2 (Dsg2) and E-cadherin (E-cad). Bar, 20 μm. (B) Staining along the border was quantified as follows: one-third border occupied (black), two-thirds (gray), or entire (white) border. TAPI- and PKI-treated cells showed increased intercellular Dsg2, but not E-cad, localization. (C) Confluent monolayers of SCC68 cells were cultured overnight in the presence of DMSO, GM6001 (GM), or PKI in 0.09, 0.50, or 1.0 mM CaCl2 . Intercellular adhesion was measured by counting the number of fragments generated from dispase-released monolayers subjected to a defined amount of mechanical stress. At 0.50 mM and 1.0 mM calcium, significantly fewer fragments were produced in the TAPI- and PKI-treated monolayers (p < 0.05).

Techniques Used: Inhibition, Immunofluorescence, Staining, Cell Culture, Generated, Produced

9) Product Images from "Liposomal Formulations for an Efficient Encapsulation of Epigallocatechin-3-Gallate: An In-Silico/Experimental Approach"

Article Title: Liposomal Formulations for an Efficient Encapsulation of Epigallocatechin-3-Gallate: An In-Silico/Experimental Approach

Journal: Molecules : A Journal of Synthetic Chemistry and Natural Product Chemistry

doi: 10.3390/molecules23020441

Representative structures for the steady states of neutral models with CaCl2 . The models without salts ( A ), 1:1 ( B ), 3:1 ( C ), 5:1 ( D ) and 6:1 ( E ) are shown. Ca2+ ions (cyan VdW spheres), Epigallocatechin-3-gallate (EGCG) molecules (green sticks), phosphorous atoms (orange VdW spheres) and lipid leaflets (as transparent surface) are highlighted.
Figure Legend Snippet: Representative structures for the steady states of neutral models with CaCl2 . The models without salts ( A ), 1:1 ( B ), 3:1 ( C ), 5:1 ( D ) and 6:1 ( E ) are shown. Ca2+ ions (cyan VdW spheres), Epigallocatechin-3-gallate (EGCG) molecules (green sticks), phosphorous atoms (orange VdW spheres) and lipid leaflets (as transparent surface) are highlighted.

Techniques Used:

10) Product Images from "Forty mouse strain survey of voluntary calcium intake, blood calcium, and bone mineral content"

Article Title: Forty mouse strain survey of voluntary calcium intake, blood calcium, and bone mineral content

Journal: Physiology & behavior

doi: 10.1016/j.physbeh.2007.03.027

Scatter plot of intake of three concentrations of CaCl2 versus calcium lactate (CaLa; r =0.75). Each point represents the mean value of ~10 mice of the same sex and strain. The four outlying high points are all from the MSM/MsJ strain. The outlying low point is from C58/J females given 7.5 mM concentrations.
Figure Legend Snippet: Scatter plot of intake of three concentrations of CaCl2 versus calcium lactate (CaLa; r =0.75). Each point represents the mean value of ~10 mice of the same sex and strain. The four outlying high points are all from the MSM/MsJ strain. The outlying low point is from C58/J females given 7.5 mM concentrations.

Techniques Used: Mouse Assay

11) Product Images from "Interaction of Heparins and Dextran Sulfates with a Mesoscopic Protein Nanopore"

Article Title: Interaction of Heparins and Dextran Sulfates with a Mesoscopic Protein Nanopore

Journal:

doi: 10.1016/j.bpj.2009.09.019

Representations of ( A ) the α HL channel, heparin, and DS used in this work, ( B ) the charge distribution at the α HL channel, and ( C and D ) the side-dependent heparin effect. ( A ) Cross section through the α HL channel (Protein Data Bank, 7AHL.pdb) embedded in membrane. Structure of the dodecameric heparin (Protein Data Bank, 1HPN.pdb; molecular mass of 3453.64 g/mol) and eventual structure of the hexadecameric DS (molecular mass of 5009.56 g/mol) were built with Chem3D (CambridgeSoft). (Abbreviations: Hep , heparin; DS , dextran sulfate.) The atoms are represented using the following color codes: O (which possesses a negative charge) in red, C in gray, H in white, N (which possesses a positive charge) in blue, and S in yellow. ( Green triangles ) Levels where the novel Cys and its charged derivatives are located in the channel structure. Scale bar is 2 nm. ( B ) The charge distribution was calculated using Coulomb calculation method (Swiss-PDBViewer, Ver. 3.7) assuming solvent ionic strength of 0.15 mol dm−3 , equal to that of the 50 mM CaCl2 solution mainly used in this study. The isopotential contouring values, equal to 1.8 kT / e , are shown ( red , positive potentials; blue , negative potentials). ( C and D ) The current decays after application of 100 mV steps to multichannel bilayers in the presence of Hep6000 at the trans ( C ) and cis ( D ) compartments of the experimental chamber. ( Red lines ) Best-fit of a single exponential function. Control traces (no PA) are shown for comparison. Concentrations of Hep6000, voltage protocols, current, and timescales are given in the figure. Note that the current inhibition is not total. All other conditions for the experiment are described in .
Figure Legend Snippet: Representations of ( A ) the α HL channel, heparin, and DS used in this work, ( B ) the charge distribution at the α HL channel, and ( C and D ) the side-dependent heparin effect. ( A ) Cross section through the α HL channel (Protein Data Bank, 7AHL.pdb) embedded in membrane. Structure of the dodecameric heparin (Protein Data Bank, 1HPN.pdb; molecular mass of 3453.64 g/mol) and eventual structure of the hexadecameric DS (molecular mass of 5009.56 g/mol) were built with Chem3D (CambridgeSoft). (Abbreviations: Hep , heparin; DS , dextran sulfate.) The atoms are represented using the following color codes: O (which possesses a negative charge) in red, C in gray, H in white, N (which possesses a positive charge) in blue, and S in yellow. ( Green triangles ) Levels where the novel Cys and its charged derivatives are located in the channel structure. Scale bar is 2 nm. ( B ) The charge distribution was calculated using Coulomb calculation method (Swiss-PDBViewer, Ver. 3.7) assuming solvent ionic strength of 0.15 mol dm−3 , equal to that of the 50 mM CaCl2 solution mainly used in this study. The isopotential contouring values, equal to 1.8 kT / e , are shown ( red , positive potentials; blue , negative potentials). ( C and D ) The current decays after application of 100 mV steps to multichannel bilayers in the presence of Hep6000 at the trans ( C ) and cis ( D ) compartments of the experimental chamber. ( Red lines ) Best-fit of a single exponential function. Control traces (no PA) are shown for comparison. Concentrations of Hep6000, voltage protocols, current, and timescales are given in the figure. Note that the current inhibition is not total. All other conditions for the experiment are described in .

Techniques Used: Inhibition

12) Product Images from "Fabrication of Freestanding Alginate Microfibers and Microstructures for Tissue Engineering Applications"

Article Title: Fabrication of Freestanding Alginate Microfibers and Microstructures for Tissue Engineering Applications

Journal:

doi: 10.1088/1758-5082/6/2/024104

CaCl2 and PIPAAm are necessary for the release of freestanding, assembled alginate microfibers (a) When alginate was micromolded onto a glass surface, hydration in a 2% CaCl2 solution enabled the alginate to crosslink, but the resultant fibers remained permanently bound to the glass surface. (b) Alternatively, alginate fibers that were micromolded onto a PIPAAm surface and hydrated with just DI water failed to crosslink and ultimately dissolved into solution. (c) CaCl2 concentrations as low as 0.5% were sufficient to crosslink the alginate and produce microfibers upon dissolution of PIPAAm. (d) A higher concentrations of 2% CaCl2 also effectively crosslinked the alginate microfibers. Scale bars are 50 μm.
Figure Legend Snippet: CaCl2 and PIPAAm are necessary for the release of freestanding, assembled alginate microfibers (a) When alginate was micromolded onto a glass surface, hydration in a 2% CaCl2 solution enabled the alginate to crosslink, but the resultant fibers remained permanently bound to the glass surface. (b) Alternatively, alginate fibers that were micromolded onto a PIPAAm surface and hydrated with just DI water failed to crosslink and ultimately dissolved into solution. (c) CaCl2 concentrations as low as 0.5% were sufficient to crosslink the alginate and produce microfibers upon dissolution of PIPAAm. (d) A higher concentrations of 2% CaCl2 also effectively crosslinked the alginate microfibers. Scale bars are 50 μm.

Techniques Used:

A schematic of the alginate microfiber and microstructure fabrication process (a) A microfabricated PDMS stamp is pressed onto an alginate or alginate-fibrinogen drop on a PIPAAm coated coverslip. This is performed on a hotplate set to 45 °C to prevent dissolution of the PIPAAm layer. (b) The alginate is heated with the PDMS stamp in conformal contact to dry the alginate onto the PIPAAm surface. (c) Removal of the PDMS stamp yields the formation of dried, micromolded alginate microfibers. (d) The alginate fibers and structures are hydrated in a 40 °C CaCl2 solution. If desirable, cells are also seeded at this step. (e) Cooling the solution below the LCST of PIPAAm (32 °C) causes the dissolution of the PIPAAm layer and the release of assembled alginate microfibers and microstructures.
Figure Legend Snippet: A schematic of the alginate microfiber and microstructure fabrication process (a) A microfabricated PDMS stamp is pressed onto an alginate or alginate-fibrinogen drop on a PIPAAm coated coverslip. This is performed on a hotplate set to 45 °C to prevent dissolution of the PIPAAm layer. (b) The alginate is heated with the PDMS stamp in conformal contact to dry the alginate onto the PIPAAm surface. (c) Removal of the PDMS stamp yields the formation of dried, micromolded alginate microfibers. (d) The alginate fibers and structures are hydrated in a 40 °C CaCl2 solution. If desirable, cells are also seeded at this step. (e) Cooling the solution below the LCST of PIPAAm (32 °C) causes the dissolution of the PIPAAm layer and the release of assembled alginate microfibers and microstructures.

Techniques Used:

13) Product Images from "Stromal Cell-Derived Factor-1α Alleviates Calcium-Sensing Receptor Activation-Mediated Ischemia/Reperfusion Injury by Inhibiting Caspase-3/Caspase-9-Induced Cell Apoptosis in Rat Free Flaps"

Article Title: Stromal Cell-Derived Factor-1α Alleviates Calcium-Sensing Receptor Activation-Mediated Ischemia/Reperfusion Injury by Inhibiting Caspase-3/Caspase-9-Induced Cell Apoptosis in Rat Free Flaps

Journal: BioMed Research International

doi: 10.1155/2018/8945850

(a) Free flap survival status on day 7 after ischemia/reperfusion (I/R). The necrotic ratio in the CaCl2 group was significantly higher than that of the other groups, while the necrotic ratios in the NPS2143 + CaCl2 and SDF-1 α + CaCl2 groups were significantly lower than that in the NS group. (b) Apoptotic cell analysis by TUNEL staining after 2 h or 7 d of reperfusion. Apoptotic cells in the CaCl2 group were remarkably increased, while apoptotic cells in the NPS2143 + CaCl2 and SDF-1 α + CaCl2 groups were notably decreased compared with the other groups. ∗∗∗ P < 0.001 versus CaCl2 group, ### P < 0.001 versus SDF-1 α + CaCl2 group, ## P < 0.01 versus SDF-1 α + CaCl2 group.
Figure Legend Snippet: (a) Free flap survival status on day 7 after ischemia/reperfusion (I/R). The necrotic ratio in the CaCl2 group was significantly higher than that of the other groups, while the necrotic ratios in the NPS2143 + CaCl2 and SDF-1 α + CaCl2 groups were significantly lower than that in the NS group. (b) Apoptotic cell analysis by TUNEL staining after 2 h or 7 d of reperfusion. Apoptotic cells in the CaCl2 group were remarkably increased, while apoptotic cells in the NPS2143 + CaCl2 and SDF-1 α + CaCl2 groups were notably decreased compared with the other groups. ∗∗∗ P < 0.001 versus CaCl2 group, ### P < 0.001 versus SDF-1 α + CaCl2 group, ## P < 0.01 versus SDF-1 α + CaCl2 group.

Techniques Used: TUNEL Assay, Staining

14) Product Images from "Transcranial manganese delivery for neuronal tract tracing using MEMRI"

Article Title: Transcranial manganese delivery for neuronal tract tracing using MEMRI

Journal:

doi: 10.1016/j.neuroimage.2017.05.025

Concentration dependence and the calcium addition effect on transcranial manganese delivery efficiency R1 -maps calculated from saturation recovery MR images of rats receiving - (A) 100 mM MnCl2 (slice shown is approximately 1.5 mm anterior from bregma), (B) 100 mM MnCl2 +400 mM CaCl2 (slice is approximately −0.6 mm posterior from bregma), (C) 250 mM MnCl2 (slice shown is approximately −0.5 mm posterior from bregma) and (D) 300 mM MnCl2 +200 mM CaCl2 (slice shown is approximately −0.6 mm posterior from bregma) on the bregma and dependence of total amount of manganese delivered to the brain on the concentration of manganese solution applied on the bregma, when only MnCl2 was applied (red diamonds) and when CaCl2 was added to the total concentration of 500 mM (blue squares) (E). Scale bar, 2 mm. R1 intensity scaling, 0.125–1 s−1 . ROIs where T1 measurements were made are shown on the top of the R1 maps in red or blue, whereas the green ROIs represent the background. Ca2+ increases the efficiency of Mn2+ delivery through the skull. Each data point represents the average of three animals.
Figure Legend Snippet: Concentration dependence and the calcium addition effect on transcranial manganese delivery efficiency R1 -maps calculated from saturation recovery MR images of rats receiving - (A) 100 mM MnCl2 (slice shown is approximately 1.5 mm anterior from bregma), (B) 100 mM MnCl2 +400 mM CaCl2 (slice is approximately −0.6 mm posterior from bregma), (C) 250 mM MnCl2 (slice shown is approximately −0.5 mm posterior from bregma) and (D) 300 mM MnCl2 +200 mM CaCl2 (slice shown is approximately −0.6 mm posterior from bregma) on the bregma and dependence of total amount of manganese delivered to the brain on the concentration of manganese solution applied on the bregma, when only MnCl2 was applied (red diamonds) and when CaCl2 was added to the total concentration of 500 mM (blue squares) (E). Scale bar, 2 mm. R1 intensity scaling, 0.125–1 s−1 . ROIs where T1 measurements were made are shown on the top of the R1 maps in red or blue, whereas the green ROIs represent the background. Ca2+ increases the efficiency of Mn2+ delivery through the skull. Each data point represents the average of three animals.

Techniques Used: Concentration Assay, Recovery

15) Product Images from "TRP Channels Localize to Subdomains of the Apical Plasma Membrane in Human Fetal Retinal Pigment Epithelium"

Article Title: TRP Channels Localize to Subdomains of the Apical Plasma Membrane in Human Fetal Retinal Pigment Epithelium

Journal:

doi: 10.1167/iovs.14-15738

Effect of BaCl2 on hfRPE in culture media. The TER is reported relative to the TER at the start of the experiment (TER ≥ 300 Ω × cm2 ). Except where noted, the results are based on three independent experiments (mean ± SE). ( A ) The results for SFM-1 and SFM-1 plus caloxin 1b1 (Ca2+ -ATPase inhibitor) were indistinguishable. The effect of adding CaCl2 was transient with a small, but significant decrease ( P < 0.05) at 2 hours. The decrease was prolonged in the presence of the inhibitor ( P < 0.05 relative to Ca2+ alone). Barium chloride caused significant reduction of TER after 2 and 4 hours in SFM at 37°C. Similar results were obtained when the TER of hfRPE was elevated by maintaining hfRPE in a serum-containing culture medium (not shown). ( B ) Light microscopy revealed normal polygonal RPE morphology after 4 hours in 5 mM CaCl2 ( left ) and loss of the apical junctional complex after 4 hours in BaCl2 ( right ). ( C ) The TER decreased when BaCl2 was added to the apical media chamber, but not when added to the basolateral chamber (mean ± SE). Valinomycin and nifedipine were ineffective at preventing TER reduction (mean ± range, n = 2). ( D ) Lanthanum chloride in the apical chamber partially blocked the Ba2+ -mediated effect, but LaCl3 in the basolateral chamber was ineffective. Inhibitors of TRPC4 (ML204) and TRPV4 (HC-067047), alone and in combination (ML/HC), failed to block the effect of Ba2+ (mean ± range, n = 2).
Figure Legend Snippet: Effect of BaCl2 on hfRPE in culture media. The TER is reported relative to the TER at the start of the experiment (TER ≥ 300 Ω × cm2 ). Except where noted, the results are based on three independent experiments (mean ± SE). ( A ) The results for SFM-1 and SFM-1 plus caloxin 1b1 (Ca2+ -ATPase inhibitor) were indistinguishable. The effect of adding CaCl2 was transient with a small, but significant decrease ( P < 0.05) at 2 hours. The decrease was prolonged in the presence of the inhibitor ( P < 0.05 relative to Ca2+ alone). Barium chloride caused significant reduction of TER after 2 and 4 hours in SFM at 37°C. Similar results were obtained when the TER of hfRPE was elevated by maintaining hfRPE in a serum-containing culture medium (not shown). ( B ) Light microscopy revealed normal polygonal RPE morphology after 4 hours in 5 mM CaCl2 ( left ) and loss of the apical junctional complex after 4 hours in BaCl2 ( right ). ( C ) The TER decreased when BaCl2 was added to the apical media chamber, but not when added to the basolateral chamber (mean ± SE). Valinomycin and nifedipine were ineffective at preventing TER reduction (mean ± range, n = 2). ( D ) Lanthanum chloride in the apical chamber partially blocked the Ba2+ -mediated effect, but LaCl3 in the basolateral chamber was ineffective. Inhibitors of TRPC4 (ML204) and TRPV4 (HC-067047), alone and in combination (ML/HC), failed to block the effect of Ba2+ (mean ± range, n = 2).

Techniques Used: Light Microscopy, Blocking Assay

Related Articles

Zymography:

Article Title: Biological and Enzymatic Characterization of Proteases from Crude Venom of the Ant Odontomachus bauri
Article Snippet: Paragraph title: 4.4.4. Gelatin Zymography ... Subsequent to the electrophoresis, the gel was washed twice for 30 min at room temperature in 2.5% Triton X-100 (Sigma-Aldrich) to remove the SDS and incubated at 37 °C for 18 h in one of the following buffers: 0.05 M sodium citrate pH 4.0, pH 5.0 and pH 6.0; 0.05 M Tris-HCl pH 7.0, pH 8.0, pH 9.0 and pH 10.0; and in the presence of ions and other chemicals as 50 mM Tris-HCl pH 8.0; 50 mM Tris-HCl and 10 mM CaCl2 (Sigma-Aldrich) pH 8.0; 50 mM Tris-HCl, 150 mM NaCl, 10 mM CaCl2 , 0.002% CHAPS (Sigma-Aldrich) and 10 mM EDTA pH 8.0; and 50 mM Tris-HCl, 1 mM CaCl2 and 1 mM ZnSO4 (Sigma-Aldrich) pH 8.0.

Electrophoresis:

Article Title: Biological and Enzymatic Characterization of Proteases from Crude Venom of the Ant Odontomachus bauri
Article Snippet: Crude venom samples (5 µg) were separated by 12% SDS-PAGE containing 1% of the gelatin substrate (Sigma-Aldrich). .. Subsequent to the electrophoresis, the gel was washed twice for 30 min at room temperature in 2.5% Triton X-100 (Sigma-Aldrich) to remove the SDS and incubated at 37 °C for 18 h in one of the following buffers: 0.05 M sodium citrate pH 4.0, pH 5.0 and pH 6.0; 0.05 M Tris-HCl pH 7.0, pH 8.0, pH 9.0 and pH 10.0; and in the presence of ions and other chemicals as 50 mM Tris-HCl pH 8.0; 50 mM Tris-HCl and 10 mM CaCl2 (Sigma-Aldrich) pH 8.0; 50 mM Tris-HCl, 150 mM NaCl, 10 mM CaCl2 , 0.002% CHAPS (Sigma-Aldrich) and 10 mM EDTA pH 8.0; and 50 mM Tris-HCl, 1 mM CaCl2 and 1 mM ZnSO4 (Sigma-Aldrich) pH 8.0. .. The gels were stained with R-250 Coomassie blue and gelatin proteolysis activity detected as colorless bands in the otherwise blue gel.

Incubation:

Article Title: E-Cadherin fragments as potential mediators for peritoneal metastasis in advanced epithelial ovarian cancer
Article Snippet: After washing in PBS, the cells were scratched with a cell scraper and homogenised in extraction buffer (50 mM HEPES (ph 7.5), 1 mM EGTA, 50 mM KCl, 2 mM MgCl2, 5 mM Mercaptoethanol and 1 μ l protease inhibitor cocktail without Leupeptin (P8849, Sigma-Aldrich) per 100 μ l extraction-buffer). .. Equal amounts of protein lysates were incubated with different treatments for 60 min at 30 °C: without adjunct, with 1 mM CaCl2 , with 1 mM CaCl2 and 0.3 U (Units) μ -Calpain (C6108, Sigma-Aldrich), and with 1 mM CaCl2 , 0.3 U μ -Calpain and 10 mM Calpeptin (CAS 117591-20-5, Calbiochem, Darmstadt, Germany). .. Treatment was stopped by addition of 2 × sample buffer (62.5 mm Tris-HCL (pH 6.8), 4% SDS, 10% glycerol) and 10-min incubation at 100 °C.

Article Title: Biological and Enzymatic Characterization of Proteases from Crude Venom of the Ant Odontomachus bauri
Article Snippet: Crude venom samples (5 µg) were separated by 12% SDS-PAGE containing 1% of the gelatin substrate (Sigma-Aldrich). .. Subsequent to the electrophoresis, the gel was washed twice for 30 min at room temperature in 2.5% Triton X-100 (Sigma-Aldrich) to remove the SDS and incubated at 37 °C for 18 h in one of the following buffers: 0.05 M sodium citrate pH 4.0, pH 5.0 and pH 6.0; 0.05 M Tris-HCl pH 7.0, pH 8.0, pH 9.0 and pH 10.0; and in the presence of ions and other chemicals as 50 mM Tris-HCl pH 8.0; 50 mM Tris-HCl and 10 mM CaCl2 (Sigma-Aldrich) pH 8.0; 50 mM Tris-HCl, 150 mM NaCl, 10 mM CaCl2 , 0.002% CHAPS (Sigma-Aldrich) and 10 mM EDTA pH 8.0; and 50 mM Tris-HCl, 1 mM CaCl2 and 1 mM ZnSO4 (Sigma-Aldrich) pH 8.0. .. The gels were stained with R-250 Coomassie blue and gelatin proteolysis activity detected as colorless bands in the otherwise blue gel.

Article Title: Residual Detergent Detection Method for Nondestructive Cytocompatibility Evaluation of Decellularized Whole Lung Scaffolds
Article Snippet: Following a 5, 15, 30, or 60 min incubation period at room temperature, 150 μL of the bottom chloroform layer was extracted ( ; Supplementary Data are available online at ) and the absorbance was measured in a Synergy HT Multi-Detection Microplate Reader (Biotek Instruments, Winooski, VT). .. To analyze the effect of other ions and reagents commonly utilized during decellularization protocols, , , , , , , , , we determined the SDC concentration in the presence of a range of Triton X-100 (0–1%), 2 mM CaCl2 (Sigma-Aldrich), 1.3 mM MgSO4 (Sigma-Aldrich), a solution of both 2 mM CaCl2 (Sigma-Aldrich) and 1.3 mM MgSO4 (Sigma-Aldrich), or 1 M NaCl (Fisher Scientific, Hampton, NH).

Article Title: TRP Channels Localize to Subdomains of the Apical Plasma Membrane in Human Fetal Retinal Pigment Epithelium
Article Snippet: Final concentrations were: 3.0 mM BaCl2 (Thermo Fisher Scientific, Rockford, IL, USA), 6.3 mM CaCl2 (Sigma-Aldrich Corp., St. Louis, MO, USA), 2.0 mM LaCl3 (Sigma-Aldrich Corp.), 5.0 μM valinomycin (R & D Systems, Minneapolis, MN, USA), 10 μM nifedipine (Alfa Aesar, Ward Hill, MA, USA), 20 μM ML204 (Sigma-Aldrich Corp.), 1.0 μM HC-067047 (Sigma-Aldrich Corp.), 100 μM ALLM (Santa Cruz Biotechnology, Dallas, TX, USA), and 400 μM caloxin 1b1. .. Final concentrations were: 3.0 mM BaCl2 (Thermo Fisher Scientific, Rockford, IL, USA), 6.3 mM CaCl2 (Sigma-Aldrich Corp., St. Louis, MO, USA), 2.0 mM LaCl3 (Sigma-Aldrich Corp.), 5.0 μM valinomycin (R & D Systems, Minneapolis, MN, USA), 10 μM nifedipine (Alfa Aesar, Ward Hill, MA, USA), 20 μM ML204 (Sigma-Aldrich Corp.), 1.0 μM HC-067047 (Sigma-Aldrich Corp.), 100 μM ALLM (Santa Cruz Biotechnology, Dallas, TX, USA), and 400 μM caloxin 1b1.

Diffusion-based Assay:

Article Title: Transcranial manganese delivery for neuronal tract tracing using MEMRI
Article Snippet: Solutions of CaCl2 , NaCl, MgCl2 and D-mannitol (Sigma-Aldrich, St. Louis, MO) in different concentrations were each separately added to some of the MnCl2 solutions to study the possible effect of ionic strength and osmolarity on the delivery efficiency. .. Solutions of CaCl2 , NaCl, MgCl2 and D-mannitol (Sigma-Aldrich, St. Louis, MO) in different concentrations were each separately added to some of the MnCl2 solutions to study the possible effect of ionic strength and osmolarity on the delivery efficiency.

Activity Assay:

Article Title: Sustained-release synthetic biomarkers for monitoring thrombosis and inflammation using point-of-care compatible readouts
Article Snippet: For plasma studies, PEG-T1Q was mixed with 50 μL of control human plasma (Thermo Scientific) and 50 μL of 80 mM CaCl2 (Sigma) to initiate the clotting cascade with or without the thrombin inhibitor (Argatroban Monohydrate, Sigma) or PBS. .. For plasma studies, PEG-T1Q was mixed with 50 μL of control human plasma (Thermo Scientific) and 50 μL of 80 mM CaCl2 (Sigma) to initiate the clotting cascade with or without the thrombin inhibitor (Argatroban Monohydrate, Sigma) or PBS.

Expressing:

Article Title: Stromal Cell-Derived Factor-1α Alleviates Calcium-Sensing Receptor Activation-Mediated Ischemia/Reperfusion Injury by Inhibiting Caspase-3/Caspase-9-Induced Cell Apoptosis in Rat Free Flaps
Article Snippet: All rats were equally randomized into Groups A, B, C, D, and E ( n = 10 per group), in which they received CaCl2 (0.1 mL/kg; Sigma-Aldrich, St. Louis, MO, USA), NPS2143 (1 mg/kg; Selleck Chemicals, Houston, TX, USA) + CaCl2 , SDF-1 α (10 mg/kg; Cyagen Biosciences, Santa Clara, CA, USA) + CaCl2 , AMD3100 (5 mg/kg; Sigma-Aldrich) + SDF-1 α + CaCl2 , and normal saline (NS) only, respectively [ ]. .. All rats were equally randomized into Groups A, B, C, D, and E ( n = 10 per group), in which they received CaCl2 (0.1 mL/kg; Sigma-Aldrich, St. Louis, MO, USA), NPS2143 (1 mg/kg; Selleck Chemicals, Houston, TX, USA) + CaCl2 , SDF-1 α (10 mg/kg; Cyagen Biosciences, Santa Clara, CA, USA) + CaCl2 , AMD3100 (5 mg/kg; Sigma-Aldrich) + SDF-1 α + CaCl2 , and normal saline (NS) only, respectively [ ].

Modification:

Article Title: C - Reactive Protein Induced Rearrangement of Phosphatidylcholine on Nanoparticle Mimics of Lipoprotein Particles
Article Snippet: Human CRP in 10 mM Tris at pH 8.0 containing 140 mM NaCl, 0.1% NaN3 , and 2.22 mM CaCl2 and 95% L-α-phosphatidylcholine (PC) were from Sigma Aldrich. .. Human CRP in 10 mM Tris at pH 8.0 containing 140 mM NaCl, 0.1% NaN3 , and 2.22 mM CaCl2 and 95% L-α-phosphatidylcholine (PC) were from Sigma Aldrich.

Article Title: Coaxial Extrusion Bioprinting of 3D Microfibrous Constructs with Cell-Favorable Gelatin Methacryloyl Microenvironments
Article Snippet: Gelatin, methacrylic anhydride, 2-hydroxy-4’-(2-hydroxyethoxy)-2-methylpropiophenone (photoinitiator, PI), alginate, and CaCl2 were purchased from Sigma-Aldrich (St. Louis, MO, USA). .. GelMA molecules were synthesized according to our previously published protocol [ , ], at a medium degree of methacryloyl substitution (53.8 ± 0.5%).

Western Blot:

Article Title: E-Cadherin fragments as potential mediators for peritoneal metastasis in advanced epithelial ovarian cancer
Article Snippet: Equal amounts of protein lysates were incubated with different treatments for 60 min at 30 °C: without adjunct, with 1 mM CaCl2 , with 1 mM CaCl2 and 0.3 U (Units) μ -Calpain (C6108, Sigma-Aldrich), and with 1 mM CaCl2 , 0.3 U μ -Calpain and 10 mM Calpeptin (CAS 117591-20-5, Calbiochem, Darmstadt, Germany). .. Equal amounts of protein lysates were incubated with different treatments for 60 min at 30 °C: without adjunct, with 1 mM CaCl2 , with 1 mM CaCl2 and 0.3 U (Units) μ -Calpain (C6108, Sigma-Aldrich), and with 1 mM CaCl2 , 0.3 U μ -Calpain and 10 mM Calpeptin (CAS 117591-20-5, Calbiochem, Darmstadt, Germany).

Magnetic Resonance Imaging:

Article Title: Transcranial manganese delivery for neuronal tract tracing using MEMRI
Article Snippet: Solutions of CaCl2 , NaCl, MgCl2 and D-mannitol (Sigma-Aldrich, St. Louis, MO) in different concentrations were each separately added to some of the MnCl2 solutions to study the possible effect of ionic strength and osmolarity on the delivery efficiency. .. Solutions of CaCl2 , NaCl, MgCl2 and D-mannitol (Sigma-Aldrich, St. Louis, MO) in different concentrations were each separately added to some of the MnCl2 solutions to study the possible effect of ionic strength and osmolarity on the delivery efficiency.

Electron Microscopy:

Article Title: Sustained-release synthetic biomarkers for monitoring thrombosis and inflammation using point-of-care compatible readouts
Article Snippet: Fluorescence was monitored using a microplate reader (either SpectroMax Gemini EM or Tecan Infinite). .. For plasma studies, PEG-T1Q was mixed with 50 μL of control human plasma (Thermo Scientific) and 50 μL of 80 mM CaCl2 (Sigma) to initiate the clotting cascade with or without the thrombin inhibitor (Argatroban Monohydrate, Sigma) or PBS.

Flow Cytometry:

Article Title: Adipose Stem Cell Microbeads as Production Sources for Chondrogenic Growth Factors
Article Snippet: Microbeads containing the commercial ASCs seeded homogenously throughout the hydrogel were created using a Nisco Encapsulator VAR V1 LIN-0043 (Nisco Engineering AG, Zurich, Switzerland) at a 10 mL/hr flow rate, 0.12 mm inner diameter nozzle and 6kV/cm electrostatic potential[ , ] . .. In studies determining growth factor production and secretion from ASC microbeads over time and in studies determining the effect of chondrogenic medium and microencapsulation on ASCs from multiple donors, microbeads with 130 kDa, 44% guluronate alginate (LVM, low viscosity high mannuronate) were crosslinked in a solution of 50 mM CaCl2 (Sigma, St. Louis, MO, USA), 150 mM glucose (Sigma) and 15 mM 4-(2-hydroxyethyl)-1-piperazineethanesulfonic acid (HEPES, Sigma) at pH 7.3 for at least 15 minutes.

Protease Inhibitor:

Article Title: E-Cadherin fragments as potential mediators for peritoneal metastasis in advanced epithelial ovarian cancer
Article Snippet: After washing in PBS, the cells were scratched with a cell scraper and homogenised in extraction buffer (50 mM HEPES (ph 7.5), 1 mM EGTA, 50 mM KCl, 2 mM MgCl2, 5 mM Mercaptoethanol and 1 μ l protease inhibitor cocktail without Leupeptin (P8849, Sigma-Aldrich) per 100 μ l extraction-buffer). .. Equal amounts of protein lysates were incubated with different treatments for 60 min at 30 °C: without adjunct, with 1 mM CaCl2 , with 1 mM CaCl2 and 0.3 U (Units) μ -Calpain (C6108, Sigma-Aldrich), and with 1 mM CaCl2 , 0.3 U μ -Calpain and 10 mM Calpeptin (CAS 117591-20-5, Calbiochem, Darmstadt, Germany).

Transferring:

Article Title: Identification of Chloride Channels CLCN3 and CLCN5 Mediating the Excitatory Cl− Currents Activated by Sphingosine-1-Phosphate in Sensory Neurons
Article Snippet: The external solution (ECS) contained (in mM): 145 NaCl, 5 KCl, 2 CaCl2 , 1 MgCl2 (all Sigma), 10 glucose and 10 HEPES (Merck, Darmstadt, Germany), at pH 7.3 adjusted with NaOH (Merck), and electrodes were filled with internal solution (ICS, in mM): 140 KCl, 2 MgCl2 , 2 Na-ATP, 0.2 Na-GTP, 0.1 CaCl2 , 1 EGTA (all Sigma) and 10 HEPES (Merck), at pH 7.3 adjusted with KOH (Merck). .. The external solution (ECS) contained (in mM): 145 NaCl, 5 KCl, 2 CaCl2 , 1 MgCl2 (all Sigma), 10 glucose and 10 HEPES (Merck, Darmstadt, Germany), at pH 7.3 adjusted with NaOH (Merck), and electrodes were filled with internal solution (ICS, in mM): 140 KCl, 2 MgCl2 , 2 Na-ATP, 0.2 Na-GTP, 0.1 CaCl2 , 1 EGTA (all Sigma) and 10 HEPES (Merck), at pH 7.3 adjusted with KOH (Merck).

other:

Article Title: Liposomal Formulations for an Efficient Encapsulation of Epigallocatechin-3-Gallate: An In-Silico/Experimental Approach
Article Snippet: Tween-20, CaCl2 and MgCl2 salts and all solvents were obtained from Sigma Aldrich (St. Louis, MO, USA).

Inverted Microscopy:

Article Title: Adipose Stem Cell Microbeads as Production Sources for Chondrogenic Growth Factors
Article Snippet: Microbeads were imaged with an inverted microscope (Motic, Richmond, British Columbia , Canada) and had a mean diameter ± standard deviation of 122±15 μm (Motic Images Plus 2.0). .. In studies determining growth factor production and secretion from ASC microbeads over time and in studies determining the effect of chondrogenic medium and microencapsulation on ASCs from multiple donors, microbeads with 130 kDa, 44% guluronate alginate (LVM, low viscosity high mannuronate) were crosslinked in a solution of 50 mM CaCl2 (Sigma, St. Louis, MO, USA), 150 mM glucose (Sigma) and 15 mM 4-(2-hydroxyethyl)-1-piperazineethanesulfonic acid (HEPES, Sigma) at pH 7.3 for at least 15 minutes.

Injection:

Article Title: Stromal Cell-Derived Factor-1α Alleviates Calcium-Sensing Receptor Activation-Mediated Ischemia/Reperfusion Injury by Inhibiting Caspase-3/Caspase-9-Induced Cell Apoptosis in Rat Free Flaps
Article Snippet: All rats were equally randomized into Groups A, B, C, D, and E ( n = 10 per group), in which they received CaCl2 (0.1 mL/kg; Sigma-Aldrich, St. Louis, MO, USA), NPS2143 (1 mg/kg; Selleck Chemicals, Houston, TX, USA) + CaCl2 , SDF-1 α (10 mg/kg; Cyagen Biosciences, Santa Clara, CA, USA) + CaCl2 , AMD3100 (5 mg/kg; Sigma-Aldrich) + SDF-1 α + CaCl2 , and normal saline (NS) only, respectively [ ]. .. All rats were equally randomized into Groups A, B, C, D, and E ( n = 10 per group), in which they received CaCl2 (0.1 mL/kg; Sigma-Aldrich, St. Louis, MO, USA), NPS2143 (1 mg/kg; Selleck Chemicals, Houston, TX, USA) + CaCl2 , SDF-1 α (10 mg/kg; Cyagen Biosciences, Santa Clara, CA, USA) + CaCl2 , AMD3100 (5 mg/kg; Sigma-Aldrich) + SDF-1 α + CaCl2 , and normal saline (NS) only, respectively [ ].

Recombinant:

Article Title: Sustained-release synthetic biomarkers for monitoring thrombosis and inflammation using point-of-care compatible readouts
Article Snippet: PEG-T1Q (1 μM by peptide) was mixed with recombinant human thrombin (~10 nM working concentration, Haematologic Technologies) in a 384-well plate at 37°C in PBS BSA (1% wt/vol). .. For plasma studies, PEG-T1Q was mixed with 50 μL of control human plasma (Thermo Scientific) and 50 μL of 80 mM CaCl2 (Sigma) to initiate the clotting cascade with or without the thrombin inhibitor (Argatroban Monohydrate, Sigma) or PBS.

Molecular Weight:

Article Title: Adipose Stem Cell Microbeads as Production Sources for Chondrogenic Growth Factors
Article Snippet: In studies determining growth factor production and secretion from ASC microbeads over time and in studies determining the effect of chondrogenic medium and microencapsulation on ASCs from multiple donors, microbeads with 130 kDa, 44% guluronate alginate (LVM, low viscosity high mannuronate) were crosslinked in a solution of 50 mM CaCl2 (Sigma, St. Louis, MO, USA), 150 mM glucose (Sigma) and 15 mM 4-(2-hydroxyethyl)-1-piperazineethanesulfonic acid (HEPES, Sigma) at pH 7.3 for at least 15 minutes. .. In studies determining growth factor production and secretion from ASC microbeads over time and in studies determining the effect of chondrogenic medium and microencapsulation on ASCs from multiple donors, microbeads with 130 kDa, 44% guluronate alginate (LVM, low viscosity high mannuronate) were crosslinked in a solution of 50 mM CaCl2 (Sigma, St. Louis, MO, USA), 150 mM glucose (Sigma) and 15 mM 4-(2-hydroxyethyl)-1-piperazineethanesulfonic acid (HEPES, Sigma) at pH 7.3 for at least 15 minutes.

Fluorescence:

Article Title: Sustained-release synthetic biomarkers for monitoring thrombosis and inflammation using point-of-care compatible readouts
Article Snippet: Fluorescence was monitored using a microplate reader (either SpectroMax Gemini EM or Tecan Infinite). .. For plasma studies, PEG-T1Q was mixed with 50 μL of control human plasma (Thermo Scientific) and 50 μL of 80 mM CaCl2 (Sigma) to initiate the clotting cascade with or without the thrombin inhibitor (Argatroban Monohydrate, Sigma) or PBS.

Isolation:

Article Title: Identification of Chloride Channels CLCN3 and CLCN5 Mediating the Excitatory Cl− Currents Activated by Sphingosine-1-Phosphate in Sensory Neurons
Article Snippet: Ionic currents were recorded from isolated DRG neurons in the whole-cell voltage-clamp configuration at −60 mV holding potential, unless otherwise indicated. .. The external solution (ECS) contained (in mM): 145 NaCl, 5 KCl, 2 CaCl2 , 1 MgCl2 (all Sigma), 10 glucose and 10 HEPES (Merck, Darmstadt, Germany), at pH 7.3 adjusted with NaOH (Merck), and electrodes were filled with internal solution (ICS, in mM): 140 KCl, 2 MgCl2 , 2 Na-ATP, 0.2 Na-GTP, 0.1 CaCl2 , 1 EGTA (all Sigma) and 10 HEPES (Merck), at pH 7.3 adjusted with KOH (Merck).

Labeling:

Article Title: Fabrication of Freestanding Alginate Microfibers and Microstructures for Tissue Engineering Applications
Article Snippet: The alginate features were released by hydration in 40 °C CaCl2 (Sigma-Aldrich) solution with concentrations ranging between 0.5-2% (w/v) ( ). .. The alginate features were released by hydration in 40 °C CaCl2 (Sigma-Aldrich) solution with concentrations ranging between 0.5-2% (w/v) ( ).

Coagulation:

Article Title: Sustained-release synthetic biomarkers for monitoring thrombosis and inflammation using point-of-care compatible readouts
Article Snippet: Fluorescence was monitored using a microplate reader (either SpectroMax Gemini EM or Tecan Infinite). .. For plasma studies, PEG-T1Q was mixed with 50 μL of control human plasma (Thermo Scientific) and 50 μL of 80 mM CaCl2 (Sigma) to initiate the clotting cascade with or without the thrombin inhibitor (Argatroban Monohydrate, Sigma) or PBS. .. For serum stability, PEG-T1Q was added to control human serum and placed at 37°C overnight and then tested for responsiveness to thrombin compared to fresh PEG-T1Q.

Microscopy:

Article Title: Fabrication of Freestanding Alginate Microfibers and Microstructures for Tissue Engineering Applications
Article Snippet: Once dried, the PDMS stamp was removed ( ) and the fidelity of the micromolded alginate microfibers and microstructures were inspected using phase contrast microscopy. .. The alginate features were released by hydration in 40 °C CaCl2 (Sigma-Aldrich) solution with concentrations ranging between 0.5-2% (w/v) ( ).

SDS Page:

Article Title: Biological and Enzymatic Characterization of Proteases from Crude Venom of the Ant Odontomachus bauri
Article Snippet: Subsequent to the electrophoresis, the gel was washed twice for 30 min at room temperature in 2.5% Triton X-100 (Sigma-Aldrich) to remove the SDS and incubated at 37 °C for 18 h in one of the following buffers: 0.05 M sodium citrate pH 4.0, pH 5.0 and pH 6.0; 0.05 M Tris-HCl pH 7.0, pH 8.0, pH 9.0 and pH 10.0; and in the presence of ions and other chemicals as 50 mM Tris-HCl pH 8.0; 50 mM Tris-HCl and 10 mM CaCl2 (Sigma-Aldrich) pH 8.0; 50 mM Tris-HCl, 150 mM NaCl, 10 mM CaCl2 , 0.002% CHAPS (Sigma-Aldrich) and 10 mM EDTA pH 8.0; and 50 mM Tris-HCl, 1 mM CaCl2 and 1 mM ZnSO4 (Sigma-Aldrich) pH 8.0. .. Subsequent to the electrophoresis, the gel was washed twice for 30 min at room temperature in 2.5% Triton X-100 (Sigma-Aldrich) to remove the SDS and incubated at 37 °C for 18 h in one of the following buffers: 0.05 M sodium citrate pH 4.0, pH 5.0 and pH 6.0; 0.05 M Tris-HCl pH 7.0, pH 8.0, pH 9.0 and pH 10.0; and in the presence of ions and other chemicals as 50 mM Tris-HCl pH 8.0; 50 mM Tris-HCl and 10 mM CaCl2 (Sigma-Aldrich) pH 8.0; 50 mM Tris-HCl, 150 mM NaCl, 10 mM CaCl2 , 0.002% CHAPS (Sigma-Aldrich) and 10 mM EDTA pH 8.0; and 50 mM Tris-HCl, 1 mM CaCl2 and 1 mM ZnSO4 (Sigma-Aldrich) pH 8.0.

Binding Assay:

Article Title: C - Reactive Protein Induced Rearrangement of Phosphatidylcholine on Nanoparticle Mimics of Lipoprotein Particles
Article Snippet: Human CRP in 10 mM Tris at pH 8.0 containing 140 mM NaCl, 0.1% NaN3 , and 2.22 mM CaCl2 and 95% L-α-phosphatidylcholine (PC) were from Sigma Aldrich. .. The aptamer was modified with Cy5 linked to the 5′ terminal residue.

In Vitro:

Article Title: Sustained-release synthetic biomarkers for monitoring thrombosis and inflammation using point-of-care compatible readouts
Article Snippet: Paragraph title: In vitro cleavage assays ... For plasma studies, PEG-T1Q was mixed with 50 μL of control human plasma (Thermo Scientific) and 50 μL of 80 mM CaCl2 (Sigma) to initiate the clotting cascade with or without the thrombin inhibitor (Argatroban Monohydrate, Sigma) or PBS.

Patch Clamp:

Article Title: Identification of Chloride Channels CLCN3 and CLCN5 Mediating the Excitatory Cl− Currents Activated by Sphingosine-1-Phosphate in Sensory Neurons
Article Snippet: Whole-cell patch-clamp recordings were performed using patch pipettes with a tip resistance of 2–4 MΩ as previously described (Camprubí-Robles et al., ). .. The external solution (ECS) contained (in mM): 145 NaCl, 5 KCl, 2 CaCl2 , 1 MgCl2 (all Sigma), 10 glucose and 10 HEPES (Merck, Darmstadt, Germany), at pH 7.3 adjusted with NaOH (Merck), and electrodes were filled with internal solution (ICS, in mM): 140 KCl, 2 MgCl2 , 2 Na-ATP, 0.2 Na-GTP, 0.1 CaCl2 , 1 EGTA (all Sigma) and 10 HEPES (Merck), at pH 7.3 adjusted with KOH (Merck).

Evaporation:

Article Title: Residual Detergent Detection Method for Nondestructive Cytocompatibility Evaluation of Decellularized Whole Lung Scaffolds
Article Snippet: To analyze the effect of other ions and reagents commonly utilized during decellularization protocols, , , , , , , , , we determined the SDC concentration in the presence of a range of Triton X-100 (0–1%), 2 mM CaCl2 (Sigma-Aldrich), 1.3 mM MgSO4 (Sigma-Aldrich), a solution of both 2 mM CaCl2 (Sigma-Aldrich) and 1.3 mM MgSO4 (Sigma-Aldrich), or 1 M NaCl (Fisher Scientific, Hampton, NH). .. To analyze the effect of other ions and reagents commonly utilized during decellularization protocols, , , , , , , , , we determined the SDC concentration in the presence of a range of Triton X-100 (0–1%), 2 mM CaCl2 (Sigma-Aldrich), 1.3 mM MgSO4 (Sigma-Aldrich), a solution of both 2 mM CaCl2 (Sigma-Aldrich) and 1.3 mM MgSO4 (Sigma-Aldrich), or 1 M NaCl (Fisher Scientific, Hampton, NH).

Concentration Assay:

Article Title: Sustained-release synthetic biomarkers for monitoring thrombosis and inflammation using point-of-care compatible readouts
Article Snippet: PEG-T1Q (1 μM by peptide) was mixed with recombinant human thrombin (~10 nM working concentration, Haematologic Technologies) in a 384-well plate at 37°C in PBS BSA (1% wt/vol). .. For plasma studies, PEG-T1Q was mixed with 50 μL of control human plasma (Thermo Scientific) and 50 μL of 80 mM CaCl2 (Sigma) to initiate the clotting cascade with or without the thrombin inhibitor (Argatroban Monohydrate, Sigma) or PBS.

Article Title: Residual Detergent Detection Method for Nondestructive Cytocompatibility Evaluation of Decellularized Whole Lung Scaffolds
Article Snippet: Pure DI-water or PBS (Mediatech, Inc., Manassas, VA) containing no detergents served as the blank for respective experiments. .. To analyze the effect of other ions and reagents commonly utilized during decellularization protocols, , , , , , , , , we determined the SDC concentration in the presence of a range of Triton X-100 (0–1%), 2 mM CaCl2 (Sigma-Aldrich), 1.3 mM MgSO4 (Sigma-Aldrich), a solution of both 2 mM CaCl2 (Sigma-Aldrich) and 1.3 mM MgSO4 (Sigma-Aldrich), or 1 M NaCl (Fisher Scientific, Hampton, NH). .. To assess the effect of PBS on assay performance, SDC and SDS standards were prepared in PBS in addition to DI water.

Article Title: Coaxial Extrusion Bioprinting of 3D Microfibrous Constructs with Cell-Favorable Gelatin Methacryloyl Microenvironments
Article Snippet: Gelatin, methacrylic anhydride, 2-hydroxy-4’-(2-hydroxyethoxy)-2-methylpropiophenone (photoinitiator, PI), alginate, and CaCl2 were purchased from Sigma-Aldrich (St. Louis, MO, USA). .. Gelatin, methacrylic anhydride, 2-hydroxy-4’-(2-hydroxyethoxy)-2-methylpropiophenone (photoinitiator, PI), alginate, and CaCl2 were purchased from Sigma-Aldrich (St. Louis, MO, USA).

Standard Deviation:

Article Title: Adipose Stem Cell Microbeads as Production Sources for Chondrogenic Growth Factors
Article Snippet: Microbeads were imaged with an inverted microscope (Motic, Richmond, British Columbia , Canada) and had a mean diameter ± standard deviation of 122±15 μm (Motic Images Plus 2.0). .. In studies determining growth factor production and secretion from ASC microbeads over time and in studies determining the effect of chondrogenic medium and microencapsulation on ASCs from multiple donors, microbeads with 130 kDa, 44% guluronate alginate (LVM, low viscosity high mannuronate) were crosslinked in a solution of 50 mM CaCl2 (Sigma, St. Louis, MO, USA), 150 mM glucose (Sigma) and 15 mM 4-(2-hydroxyethyl)-1-piperazineethanesulfonic acid (HEPES, Sigma) at pH 7.3 for at least 15 minutes.

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    Millipore cacl2
    Cacl2, supplied by Millipore, used in various techniques. Bioz Stars score: 78/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    79
    Millipore reverse zymography digestion buffer
    The activities of MMPs and TIMPs, and cell invasion through Mtrigel are examined in hypoxic condition. Left; Gelatin and reverse <t>zymography</t> reveal that the activities of MMP-2, MMP-9 and TIMP-1 are not affected by incubation of HP-75 cells under hypoxic
    Reverse Zymography Digestion Buffer, supplied by Millipore, used in various techniques. Bioz Stars score: 79/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Millipore ip buffer
    The activities of MMPs and TIMPs, and cell invasion through Mtrigel are examined in hypoxic condition. Left; Gelatin and reverse <t>zymography</t> reveal that the activities of MMP-2, MMP-9 and TIMP-1 are not affected by incubation of HP-75 cells under hypoxic
    Ip Buffer, supplied by Millipore, used in various techniques. Bioz Stars score: 99/100, based on 44 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    The activities of MMPs and TIMPs, and cell invasion through Mtrigel are examined in hypoxic condition. Left; Gelatin and reverse zymography reveal that the activities of MMP-2, MMP-9 and TIMP-1 are not affected by incubation of HP-75 cells under hypoxic

    Journal:

    Article Title: Elevated Cell Invasion Is Induced by Hypoxia in a Human Pituitary Adenoma Cell Line

    doi:

    Figure Lengend Snippet: The activities of MMPs and TIMPs, and cell invasion through Mtrigel are examined in hypoxic condition. Left; Gelatin and reverse zymography reveal that the activities of MMP-2, MMP-9 and TIMP-1 are not affected by incubation of HP-75 cells under hypoxic

    Article Snippet: The cell supernatants were resolved as described above, and the gels were washed in 50 mM Tris, 5 mM CaCl2 and 2.5% (v/v) Triton X-100 for 2.5 hours at 23°C and then incubated in reverse zymography digestion buffer (50 mM Tris, 5 mM CaCl2 , 100 mU/ml gelatinase A (Calbiochem, Cambridge, Mass., USA) for four hours at 37°C.

    Techniques: Zymography, Incubation