dsdna (Worthington Biochemical)

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Neonatal Cardiomyocyte Isolation System

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Kit for performing five separate tissue dissociations each containing up to twelve hearts Contains single use vials of purified collagenase and trypsin CMF HBSS Leibovitz L 15 media and Falcon cell strainers along with a detailed protocol The kit is use tested by Worthington to assure performance

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lk003300

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256

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1 kt

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Worthington Biochemical dsdna
$TCR revision upon repeated immunization with antigen. (A) TCR CDR3 length profiles of mice immunized 8× with PBS, 2× or 8× with SEB. TCR repertoire of splenic CD4 + T cell was skewed only after immunization 8× with SEB. (B) Expression of V(D)J recombinase complex and related molecules in the spleen of PBS- or SEB-injected BALB/c mice. (C) GFP + cells in the Vβ8 + CD4 + T population of rag1/gfp knock-in mice. <t>IgG-RF</t> as induced in rag1/gfp knock-in mice after immunization 8× with SEB (lower left). The GFP + T cell fraction was also increased among Vβ8 + CD4 + T cells (mean ± SD, 4–5 mice/group). (D) TCRα chain revision in the spleen of mice immunized 8× with SEB was determined by LM-PCR detection of <t>dsDNA</t> breaks at the RSS flanking the TCRAV2 , with PCR-amplified TCRα constant region ( TCRAC ) as a DNA quality control.$
Kit for performing five separate tissue dissociations each containing up to twelve hearts Contains single use vials of purified collagenase and trypsin CMF HBSS Leibovitz L 15 media and Falcon cell strainers along with a detailed protocol The kit is use tested by Worthington to assure performance
https://www.bioz.com/result/dsdna/product/Worthington Biochemical
Average 92 stars, based on 194 article reviews
Price from $9.99 to $1999.99

dsdna - by Bioz Stars, 2020-10

92/100 stars

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Images

1) Product Images from "Self-Organized Criticality Theory of Autoimmunity"

Article Title: Self-Organized Criticality Theory of Autoimmunity

Journal: PLoS ONE

doi: 10.1371/journal.pone.0008382

$TCR revision upon repeated immunization with antigen. (A) TCR CDR3 length profiles of mice immunized 8× with PBS, 2× or 8× with SEB. TCR repertoire of splenic CD4 + T cell was skewed only after immunization 8× with SEB. (B) Expression of V(D)J recombinase complex and related molecules in the spleen of PBS- or SEB-injected BALB/c mice. (C) GFP + cells in the Vβ8 + CD4 + T population of rag1/gfp knock-in mice. IgG-RF as induced in rag1/gfp knock-in mice after immunization 8× with SEB (lower left). The GFP + T cell fraction was also increased among Vβ8 + CD4 + T cells (mean ± SD, 4–5 mice/group). (D) TCRα chain revision in the spleen of mice immunized 8× with SEB was determined by LM-PCR detection of dsDNA breaks at the RSS flanking the TCRAV2 , with PCR-amplified TCRα constant region ( TCRAC ) as a DNA quality control.$
Figure Legend Snippet: TCR revision upon repeated immunization with antigen. (A) TCR CDR3 length profiles of mice immunized 8× with PBS, 2× or 8× with SEB. TCR repertoire of splenic CD4 + T cell was skewed only after immunization 8× with SEB. (B) Expression of V(D)J recombinase complex and related molecules in the spleen of PBS- or SEB-injected BALB/c mice. (C) GFP + cells in the Vβ8 + CD4 + T population of rag1/gfp knock-in mice. IgG-RF as induced in rag1/gfp knock-in mice after immunization 8× with SEB (lower left). The GFP + T cell fraction was also increased among Vβ8 + CD4 + T cells (mean ± SD, 4–5 mice/group). (D) TCRα chain revision in the spleen of mice immunized 8× with SEB was determined by LM-PCR detection of dsDNA breaks at the RSS flanking the TCRAV2 , with PCR-amplified TCRα constant region ( TCRAC ) as a DNA quality control.

Techniques Used: Mouse Assay, Expressing, Injection, Knock-In, Polymerase Chain Reaction, Amplification

2) Product Images from "Carbonium vs. carbenium ion-like transition state geometries for carbocation cyclization - how strain associated with bridging affects 5-exo vs. 6-endo selectivity"

Article Title: Carbonium vs. carbenium ion-like transition state geometries for carbocation cyclization - how strain associated with bridging affects 5-exo vs. 6-endo selectivity

Journal: Chemical science (Royal Society of Chemistry : 2010)

doi: 10.1039/C3SC51657A

Computed reaction enthalpies (free energies in parentheses; kcal mol −1 ; in DCE) and barriers for [Pt(PH 3 ) 3 ] 2+ and [PdCl 2 (NCMe)] promoted 6- endo and 5- exo cyclizations. Black are for R 1 = R 2 = R 3 = CH 3 ; blue are for R1 = R3 = CH 3 /R2 = H; red are

Figure Legend Snippet: Computed reaction enthalpies (free energies in parentheses; kcal mol −1 ; in DCE) and barriers for [Pt(PH 3 ) 3 ] 2+ and [PdCl 2 (NCMe)] promoted 6- endo and 5- exo cyclizations. Black are for R 1 = R 2 = R 3 = CH 3 ; blue are for R1 = R3 = CH 3 /R2 = H; red are

Techniques Used:

3) Product Images from "Self-Organized Criticality Theory of Autoimmunity"

Article Title: Self-Organized Criticality Theory of Autoimmunity

Journal: PLoS ONE

doi: 10.1371/journal.pone.0008382

Techniques Used: Mouse Assay, Expressing, Injection, Knock-In, Polymerase Chain Reaction, Amplification

4) Product Images from "Palmitoylation Regulates Intracellular Trafficking of ?2 Adrenergic Receptor/Arrestin/Phosphodiesterase 4D Complexes in Cardiomyocytes"

Article Title: Palmitoylation Regulates Intracellular Trafficking of ?2 Adrenergic Receptor/Arrestin/Phosphodiesterase 4D Complexes in Cardiomyocytes

Journal: PLoS ONE

doi: 10.1371/journal.pone.0042658

Mutation of palmitoylation site does not inhibit agonist-induced β 2 AR trafficking. A) β 1 β 2 AR-KO cardiomyocytes expressing wild type or mutant flag-β 2 ARs were stimulated with 10 µM isoproterenol to examine receptor internalization or followed by agonist removal to examine receptor recycling. Receptors were stained by anti-flag antibody M1 and Alex 488-conjugated goat-anti-mouse secondary antibody. Maximal internalization of different flag-β 2 ARs in β 1 β 2 AR-KO cardiomyocytes at 30 minute of stimulation was plotted in bar graph. Receptors remaining on the cell surface after internalization (B and D) or recycling (C) were quantified using Alex 488-conjugated flag antibody labeling. Quantitative measurements of cell surface receptor were average from at least three different experiments. *, p

Figure Legend Snippet: Mutation of palmitoylation site does not inhibit agonist-induced β 2 AR trafficking. A) β 1 β 2 AR-KO cardiomyocytes expressing wild type or mutant flag-β 2 ARs were stimulated with 10 µM isoproterenol to examine receptor internalization or followed by agonist removal to examine receptor recycling. Receptors were stained by anti-flag antibody M1 and Alex 488-conjugated goat-anti-mouse secondary antibody. Maximal internalization of different flag-β 2 ARs in β 1 β 2 AR-KO cardiomyocytes at 30 minute of stimulation was plotted in bar graph. Receptors remaining on the cell surface after internalization (B and D) or recycling (C) were quantified using Alex 488-conjugated flag antibody labeling. Quantitative measurements of cell surface receptor were average from at least three different experiments. *, p

Techniques Used: Mutagenesis, Expressing, Staining, Antibody Labeling, Cell Surface Receptor Assay

Palmitoylation of cysteine 341 of β 2 AR is required for agonist-induced recruitment of β arrestin 2, but not receptor phosphorylation by PKA and GRK. β 1 β 2 AR-KO cardiomyocytes expressing flag-β 2 ARs and β arrestin 2-GFP were serum-starved for 30 minutes before stimulation with 10 µM of isoproterenol as indicated. (A) Cell lysates were incubated with anti-flag M2 beads to immunoprecipitate flag-β 2 ARs. The bound proteins were detected in Western blot by anti-flag and anti-GFP antibodies. *, p

Figure Legend Snippet: Palmitoylation of cysteine 341 of β 2 AR is required for agonist-induced recruitment of β arrestin 2, but not receptor phosphorylation by PKA and GRK. β 1 β 2 AR-KO cardiomyocytes expressing flag-β 2 ARs and β arrestin 2-GFP were serum-starved for 30 minutes before stimulation with 10 µM of isoproterenol as indicated. (A) Cell lysates were incubated with anti-flag M2 beads to immunoprecipitate flag-β 2 ARs. The bound proteins were detected in Western blot by anti-flag and anti-GFP antibodies. *, p

Techniques Used: Expressing, Incubation, Western Blot

$Mutation of palmitoylation site promotes caveolin-dependent β 2 AR internalization upon agonist stimulation. A) Membrane fractionations of β 1 β 2 AR-KO cardiomyocytes expressing β 2 AR or β 2 AR-C341A were blotted with caveolin-3, Src, and flag antibodies. P is the pellet fraction after centrifugation. B) Cell lysates were incubated with M2 beads for immunoprecipitation of flag-β 2 ARs to detect endogenous caveolin-3 association. (C–F) HEK293 cells expressing β 2 AR or together with other proteins were stimulated with 10 µM isoproterenol for 30 minutes to examine receptor internalization. C) Cells expressing flag-β 2 AR alone, D) cells expressing flag-β 2 ARs together dominant negative dynamin K44E, F) cell expressing flag-β 2 ARs were treated with 2 µg/ml filipin for 30 minutes, and E) cells expressing flag-β 2 AR-C341A together with β arrestin 2 (β arr2) or dominant negative β arrestin 2-V54D (β arr2-V54D), before stimulation with isoproterenol 10 µM for 30 minutes to examine receptor internalization. Cells were fixed, and receptors were stained by anti-flag antibody M1 and Alex 594-conjugated goat-anti-mouse secondary antibody.$
Figure Legend Snippet: Mutation of palmitoylation site promotes caveolin-dependent β 2 AR internalization upon agonist stimulation. A) Membrane fractionations of β 1 β 2 AR-KO cardiomyocytes expressing β 2 AR or β 2 AR-C341A were blotted with caveolin-3, Src, and flag antibodies. P is the pellet fraction after centrifugation. B) Cell lysates were incubated with M2 beads for immunoprecipitation of flag-β 2 ARs to detect endogenous caveolin-3 association. (C–F) HEK293 cells expressing β 2 AR or together with other proteins were stimulated with 10 µM isoproterenol for 30 minutes to examine receptor internalization. C) Cells expressing flag-β 2 AR alone, D) cells expressing flag-β 2 ARs together dominant negative dynamin K44E, F) cell expressing flag-β 2 ARs were treated with 2 µg/ml filipin for 30 minutes, and E) cells expressing flag-β 2 AR-C341A together with β arrestin 2 (β arr2) or dominant negative β arrestin 2-V54D (β arr2-V54D), before stimulation with isoproterenol 10 µM for 30 minutes to examine receptor internalization. Cells were fixed, and receptors were stained by anti-flag antibody M1 and Alex 594-conjugated goat-anti-mouse secondary antibody.

Techniques Used: Mutagenesis, Expressing, Centrifugation, Incubation, Immunoprecipitation, Dominant Negative Mutation, Staining

Mutation of palmitoylation reduces β 2 AR/G protein-dependent cAMP production in cardiomyocytes. β 1 β 2 AR-KO cardiomyocytes were infected with adenoviruses expressing β 2 ARs together with the plasma membrane-targeted cAMP FRET biosensor PM-ICUE3 (A and B) or cytoplasmic cAMP FRET biosensor ICUE3 (C and D). Cells were stimulated with 10 nM of isoproterenol (A and C) or 10 µM of isoproterenol (B and D). The maximal increases in FRET ratio were plotted. Rol, rolipram, was added together with Iso at 10 µM. Bar graphs were average from the maximal increases in time courses. *, p

Figure Legend Snippet: Mutation of palmitoylation reduces β 2 AR/G protein-dependent cAMP production in cardiomyocytes. β 1 β 2 AR-KO cardiomyocytes were infected with adenoviruses expressing β 2 ARs together with the plasma membrane-targeted cAMP FRET biosensor PM-ICUE3 (A and B) or cytoplasmic cAMP FRET biosensor ICUE3 (C and D). Cells were stimulated with 10 nM of isoproterenol (A and C) or 10 µM of isoproterenol (B and D). The maximal increases in FRET ratio were plotted. Rol, rolipram, was added together with Iso at 10 µM. Bar graphs were average from the maximal increases in time courses. *, p

Techniques Used: Mutagenesis, Infection, Expressing

Palmitoylation of β 2 AR is required for agonist-dependent recruitment of arrestin-PDE4D complexes to the receptor. β 1 β 2 AR-KO cardiomyocytes expressing flag-β 2 ARs together with GFP-PDE4D5 (A), GFP-PDE4D8 (B), or GFP-PDE4D9 (C) were serum-starved for 30 minutes before stimulation with 10 µM of isoproterenol as indicated. Cell lysates were incubated with anti-flag M2 beads for co-immunoprecipitation of β 2 ARs and PDE4D proteins. The bound proteins were detected in Western blot by anti-flag and anti-GFP antibodies as indicated. D) Flag-β 2 ARs and PDE4D5-RFP were expressed in wild type and arrestins double knockout (KO) MEF cells. Co-IP was done like those in panel (A–C). The bound proteins were detected in Western blot by anti-flag and anti-RFP antibodies as indicated. Western blot was quantified and normalized against the IP protein. Bar graphs were average from 3–5 different experiments. *, p

Figure Legend Snippet: Palmitoylation of β 2 AR is required for agonist-dependent recruitment of arrestin-PDE4D complexes to the receptor. β 1 β 2 AR-KO cardiomyocytes expressing flag-β 2 ARs together with GFP-PDE4D5 (A), GFP-PDE4D8 (B), or GFP-PDE4D9 (C) were serum-starved for 30 minutes before stimulation with 10 µM of isoproterenol as indicated. Cell lysates were incubated with anti-flag M2 beads for co-immunoprecipitation of β 2 ARs and PDE4D proteins. The bound proteins were detected in Western blot by anti-flag and anti-GFP antibodies as indicated. D) Flag-β 2 ARs and PDE4D5-RFP were expressed in wild type and arrestins double knockout (KO) MEF cells. Co-IP was done like those in panel (A–C). The bound proteins were detected in Western blot by anti-flag and anti-RFP antibodies as indicated. Western blot was quantified and normalized against the IP protein. Bar graphs were average from 3–5 different experiments. *, p

Techniques Used: Expressing, Incubation, Immunoprecipitation, Western Blot, Double Knockout, Co-Immunoprecipitation Assay

Stimulation of palmitoylation mutant β 2 AR leads to higher and sustained intracellular cAMP-PKA activities and higher increases in cardiomyocyte contraction rate. β 1 β 2 AR-KO cardiomyocytes expressing β 2 ARs via adenovirus infection were stimulated with isoproterenol (10 µM). The spontaneous contraction rate was recorded before and after isoproterenol stimulation. A) Quantification of isoproterenol-induced cardiomyocyte contraction rate increases over the baseline levels was plotted in the absence or presence of treatment with 2-bromopalmitic acid. B) 20 µg membrane protein containing β 2 ARs was determined by radioligand [ 3 H] DHA binding to measure expression levels of β 2 AR and β 2 AR-C341A. The expressed receptors were also detected by Western blot using anti-flag antibody. β 1 β 2 AR-KO cardiomyocytes expressing β 2 ARs together with cytoplasmic cAMP FRET biosensor ICUE3 (C-E) or PKA activity biosensor AKAR 2.2 (F). (C) and (D) Cells were stimulated with isoproterenol (10 µM) to record the changes of FRET ratio. 2-Bro, 2-bromopalmitic acid (100 µM) was added 15 minutes before Iso stimulation; Quantification of isoproterenol-induced maximal increases in cAMP FRET ratio (E) and PKA FRET ratio (F) was plotted. In each panel, bar graphs were average from the maximal increases in the indicated number of cells (FRET assay) or dishes (beating assay) from at least three independent experiments. *, p

Figure Legend Snippet: Stimulation of palmitoylation mutant β 2 AR leads to higher and sustained intracellular cAMP-PKA activities and higher increases in cardiomyocyte contraction rate. β 1 β 2 AR-KO cardiomyocytes expressing β 2 ARs via adenovirus infection were stimulated with isoproterenol (10 µM). The spontaneous contraction rate was recorded before and after isoproterenol stimulation. A) Quantification of isoproterenol-induced cardiomyocyte contraction rate increases over the baseline levels was plotted in the absence or presence of treatment with 2-bromopalmitic acid. B) 20 µg membrane protein containing β 2 ARs was determined by radioligand [ 3 H] DHA binding to measure expression levels of β 2 AR and β 2 AR-C341A. The expressed receptors were also detected by Western blot using anti-flag antibody. β 1 β 2 AR-KO cardiomyocytes expressing β 2 ARs together with cytoplasmic cAMP FRET biosensor ICUE3 (C-E) or PKA activity biosensor AKAR 2.2 (F). (C) and (D) Cells were stimulated with isoproterenol (10 µM) to record the changes of FRET ratio. 2-Bro, 2-bromopalmitic acid (100 µM) was added 15 minutes before Iso stimulation; Quantification of isoproterenol-induced maximal increases in cAMP FRET ratio (E) and PKA FRET ratio (F) was plotted. In each panel, bar graphs were average from the maximal increases in the indicated number of cells (FRET assay) or dishes (beating assay) from at least three independent experiments. *, p

Techniques Used: Mutagenesis, Expressing, Infection, Binding Assay, Western Blot, Activity Assay

Isolation:

Article Title: Construction of Cardiac Tissue Rings Using a Magnetic Tissue Fabrication Technique
Article Snippet: .. Primary Culture of Neonatal Rat Cardiomyocytes Primary neonatal rat cardiomyocytes were isolated using a neonatal cardiomyocyte isolation kit (Worthington Biochemical, Lakewood, NJ, USA) according to the published procedure [ ]. .. Briefly, ventricles from 2–4 day-old Sprague-Dawley rats (Japan SLC, Inc., Hamamatsu, Japan) were incubated for 16–20 h at 4 °C in Hank’s balanced salt solution containing trypsin (50–100 μg/mL), followed by digestion with collagenase (75 U/mL) for 40 min at 37 °C.

Article Title: Mitoregulin: A lncRNA-Encoded Microprotein that Supports Mitochondrial Supercomplexes and Respiratory Efficiency
Article Snippet: .. Neonatal rat cardiomyocytes (NRCMs) were isolated using the Worthington Neonatal Cardiomyocyte Isolation System (Worthington Biochemical Corporation, Cat. No ). ..

Article Title: Single Cell Transcriptomics Reconstructs Fate Conversion from Fibroblast to Cardiomyocyte
Article Snippet: .. Neonatal CMs were isolated using the neonatal cardiomyocytes isolation system (Worthington Biochemical Corporation) except that all enzymes were used at a ¼ of the recommended concentration to increase cell viability. .. After a 1.5 hour pre-plating on uncoated surface to remove attached nonmyocytes, the unattached CMs were collected in TRIzol ( > 80% viability by Trypan blue staining).

Article Title: ADAP1 limits neonatal cardiomyocyte hypertrophy by reducing integrin cell surface expression
Article Snippet: .. RNVC were extracted from the hearts of Sprague Dawley rat pups (strain code 001; Charles River) 1–3 days after birth using the Neonatal Cardiomyocyte Isolation System (Worthington Biochemical Corporation). .. The isolated ventricles were incubated in calcium/magnesium-free Hank’s Balanced Salt Solution containing trypsin (50 µg/mL) in vented-cap tubes with slow agitation at 4 °C for 18 h. The trypsin was then inhibited by adding Soybean Trypsin Inhibitor (200 µg/mL), and the ventricles were further digested with collagenase (100 units/mL) at 37 °C for 30 min.

Article Title: Pro-survival function of MEF2 in cardiomyocytes is enhanced by β-blockers
Article Snippet: .. Cell culture Primary neonatal rat cardiomyocytes were prepared from 1- to 3-day old Sprague Dawley rats using the Neonatal Cardiomyocyte Isolation System (Worthington Biochemical Corp, Lakewood, NJ, USA). .. Briefly, whole hearts were dissociated with trypsin (Promega, Madison, WI, USA) and collagenase (Worthington Biochemical Corp).

Article Title: Docosahexaenoic acid inhibits protein kinase C translocation/activation and cardiac hypertrophy in rat cardiomyocytes
Article Snippet: .. Isolation of cardiomyocytes Neonatal cardiomyocytes were obtained using an isolation system from Worthington Biochemical Corporation. ..

Article Title: Muscle ring finger protein-1 inhibits PKC? activation and prevents cardiomyocyte hypertrophy
Article Snippet: .. NRVM were isolated using the neonatal cardiomyocyte isolation kit (Worthington) and were plated on laminin. ..

Article Title: Pivotal role of cardiac lineage protein-1 (CLP-1) in transcriptional elongation factor P-TEFb complex formation in cardiac hypertrophy
Article Snippet: .. Cardiac cells of 2–4-day-old WKY rats were isolated using a neonatal cardiomyocyte isolation system (Worthington Biochemical Corp, Lakewood, New Jersey, USA). .. After pre-plating to reduce contaminating non-muscle cells, cardiomyocytes were plated on Bioflex collagen 6-well plates (Flexcell, McKeesport, Pennsylvania, USA) and cultured in DMEM/F-12 (1:1) media supplemented with 10% fetal bovine serum for approximately 1.5 days prior to switching to serum-free medium.

dsdna (Worthington Biochemical)

Structured Review

Related Products / Commonly Used Together

Images

1) Product Images from "Self-Organized Criticality Theory of Autoimmunity"

2) Product Images from "Carbonium vs. carbenium ion-like transition state geometries for carbocation cyclization - how strain associated with bridging affects 5-exo vs. 6-endo selectivity"

3) Product Images from "Self-Organized Criticality Theory of Autoimmunity"

4) Product Images from "Palmitoylation Regulates Intracellular Trafficking of ?2 Adrenergic Receptor/Arrestin/Phosphodiesterase 4D Complexes in Cardiomyocytes"

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