mouse ventricular cardiomyocytes (Worthington Biochemical)


Name:
Neonatal Cardiomyocyte Isolation System
Description:
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
Catalog Number:
lk003300
Price:
256
Size:
1 kt
Cas Number:
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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
https://www.bioz.com/result/mouse ventricular cardiomyocytes/product/Worthington Biochemical
Average 86 stars, based on 244 article reviews
Price from $9.99 to $1999.99
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1) Product Images from "Depletion of PHD3 Protects Heart from Ischemia/Reperfusion Injury by Inhibiting Cardiomyocyte Apoptosis"
Article Title: Depletion of PHD3 Protects Heart from Ischemia/Reperfusion Injury by Inhibiting Cardiomyocyte Apoptosis
Journal: Journal of molecular and cellular cardiology
doi: 10.1016/j.yjmcc.2015.01.007

Figure Legend Snippet: PHD3 plays a crucial role in DNA damage response and apoptosis induced by H 2 O 2 or hypoxia-reoxygenation in cardiomyocytes (A) HL-1 cells were pre-treated with DMOG for 4h or KU55933 for 30 minutes and then treated with 100 μM or 200 μM H 2 O 2 for 1 hour as indicated. Western blots were performed with the indicated antibodies. (B) HL-1 cells were cultured in a hypoxia chamber for 6 hours and then switched to normoxic conditions for the indicated time with or without pretreatment with DMOG. Western blots were performed with the indicated antibodies. (C) Neonatal mouse ventricular myocytes (NMVMs) from PHD3 f / f ; Cre +/− or PHD3 f / f ; Cre −/− mice were treated with 4-hydroxyl-tamoxifen for 3 days. Cells were then treated with NCS, Doxorubicin or H 2 O 2 for 1h and western blots were performed with indicated antibodies. (D), (E) NMVMs from PHD3 f / f mice were infected with adenovirus expressing cre recombinase or lacZ for 2 days. Infected cells were then cultured in ischemic medium at hypoxic condition for 1 hour. Re-oxygenation was obtained by culturing cells in normal medium at normoxic condition for 16 hours. Cells were immunostained with MF20 and apoptosis was analyzed with TUNEL staining. Quantitative analysis was shown in (E) . n=3, *p
Techniques Used: Western Blot, Cell Culture, Mouse Assay, Infection, Expressing, TUNEL Assay, Staining

Figure Legend Snippet: DMOG inhibits DNA damage response and apoptosis induced by doxorubicin in primary cardiomyocytes (A) Neonatal rat ventricular myocytes were pre-treated with DMOG for 4h and then treated with doxorubicin (1μM) as indicated. Western blots were performed with the indicated antibodies. (B), (C) Neonatal rat ventricular myocytes were treated with doxorubicin for 16h with or without pretreatment of DMOG. Cardiomyocyte apoptosis was then analyzed with TUNEL staining. Neonatal rat ventricular myocytes were also immunostained with MF20 antibody, which specifically recognizes myosin of striated muscle cells. Quantitative analysis is from 3 independent experiments. *p
Techniques Used: Western Blot, TUNEL Assay, Staining

Figure Legend Snippet: Depletion of PHD3 inhibits cardiomyocyte apoptosis induced by I/R injury After 5 doses of tamoxifen infusion, left anterior descending (LAD) coronary arteries of mice with indicated genotypes were tied for 40 minutes and then released for reperfusion. Twenty-four hours after reperfusion, hearts were fixed with 10% formaldehyde and embedded in paraffin. Cross sections of hearts were then analyzed with TUNEL staining. Nuclei were stained with DAPI. Representative high magnification images of the AAR are shown in (A) and low magnification images of whole sections are shown in (B) . Quantitative analysis of the apoptotic cells within the AAR is shown in (C). The numbers of mice analyzed are indicated in the bars respectively. *, p
Techniques Used: Mouse Assay, TUNEL Assay, Staining

Figure Legend Snippet: Depletion of PHD3 has no effect on HIF-1α protein level, the expression of HIF target genes or capillary density in the heart After 5 doses of tamoxifen, ventricles were excised and flash frozen. (A) Proteins extracted from ventricles of the indicated genotypes (n=3) were western-blotted with anti-HIF-1α and anti-PHD3 antibodies. Lysate from cardiomyocytes treated with DMOG (1mM) was used as the positive control for HIF-1α. (B) mRNAs were extracted from ventricles of the indicated genotypes. Relative mRNA level of HIF target genes and PHD3 were analyzed by quantitative real-time PCR. n = 3. (C) Heart sections of the indicated genotypes were immunostained with anti-myosin antibody and TRITC-E-lectin. Capillary densities are expressed as the number of lectin-positive objects per field of view. N.S., not significant, n = 3.
Techniques Used: Expressing, Western Blot, Positive Control, Real-time Polymerase Chain Reaction

Figure Legend Snippet: Depletion of PHD3 further stabilizes HIF-1α and overexpression of normoxia-stable HIF-1α protects cardiomyocytes from hypoxia-induced apoptosis (A) Neonatal ventricular myocytes from PHD3 f / f ; Cre +/− or PHD3 f / f ; Cre −/− mice were treated with 4-hydroxyl-tamoxifen for 3 days to delete PHD3. Cells were then cultured at 0.5% or 21% O 2 condition for 8 hours. Cells were then harvested for western blots with the indicated antibodies. (B), (C) HL-1 cardiomyocytes were infected with adenovirus expressing normoxia-stable HIF-1α-GFP (HIF-1α PP/AG -GFP) or lacZ for 24 hours. Infected cells were then cultured with fresh serum-free medium at 0.5% or 21% O 2 conditions for additional 48 hours to induce apoptosis. Cardiomyocytes were then fixed and stained with DAPI. Apoptosis was then analyzed with TUNEL staining. *, p
Techniques Used: Over Expression, Mouse Assay, Cell Culture, Western Blot, Infection, Expressing, Staining, TUNEL Assay

Figure Legend Snippet: DMOG inhibits HL-1 cardiomyocyte apoptosis induced by doxorubicin (A) HL-1 cells were transfected with two sets of si-RNA for Chk1 or scramble si-RNA as the control (Si-C) for two days. Cells were then treated with doxorubicin (1μM) as indicated. Western blots were then performed with the indicated antibodies. (B) After two days transfection with si-RNAs, HL-1 cells were treated with doxorubicin (1μM) for 8 hours and then harvested for caspase3/7 activity assay. Knocking down the expression of Chk-1 significantly inhibits caspase3/7 activity. N = 3, *p
Techniques Used: Transfection, Western Blot, Activity Assay, Expressing
2) Product Images from "Depletion of PHD3 Protects Heart from Ischemia/Reperfusion Injury by Inhibiting Cardiomyocyte Apoptosis"
Article Title: Depletion of PHD3 Protects Heart from Ischemia/Reperfusion Injury by Inhibiting Cardiomyocyte Apoptosis
Journal: Journal of molecular and cellular cardiology
doi: 10.1016/j.yjmcc.2015.01.007

Figure Legend Snippet: PHD3 plays a crucial role in DNA damage response and apoptosis induced by H 2 O 2 or hypoxia-reoxygenation in cardiomyocytes (A) HL-1 cells were pre-treated with DMOG for 4h or KU55933 for 30 minutes and then treated with 100 μM or 200 μM H 2 O 2 for 1 hour as indicated. Western blots were performed with the indicated antibodies. (B) HL-1 cells were cultured in a hypoxia chamber for 6 hours and then switched to normoxic conditions for the indicated time with or without pretreatment with DMOG. Western blots were performed with the indicated antibodies. (C) Neonatal mouse ventricular myocytes (NMVMs) from PHD3 f / f ; Cre +/− or PHD3 f / f ; Cre −/− mice were treated with 4-hydroxyl-tamoxifen for 3 days. Cells were then treated with NCS, Doxorubicin or H 2 O 2 for 1h and western blots were performed with indicated antibodies. (D), (E) NMVMs from PHD3 f / f mice were infected with adenovirus expressing cre recombinase or lacZ for 2 days. Infected cells were then cultured in ischemic medium at hypoxic condition for 1 hour. Re-oxygenation was obtained by culturing cells in normal medium at normoxic condition for 16 hours. Cells were immunostained with MF20 and apoptosis was analyzed with TUNEL staining. Quantitative analysis was shown in (E) . n=3, *p
Techniques Used: Western Blot, Cell Culture, Mouse Assay, Infection, Expressing, TUNEL Assay, Staining

Figure Legend Snippet: DMOG inhibits DNA damage response and apoptosis induced by doxorubicin in primary cardiomyocytes (A) Neonatal rat ventricular myocytes were pre-treated with DMOG for 4h and then treated with doxorubicin (1μM) as indicated. Western blots were performed with the indicated antibodies. (B), (C) Neonatal rat ventricular myocytes were treated with doxorubicin for 16h with or without pretreatment of DMOG. Cardiomyocyte apoptosis was then analyzed with TUNEL staining. Neonatal rat ventricular myocytes were also immunostained with MF20 antibody, which specifically recognizes myosin of striated muscle cells. Quantitative analysis is from 3 independent experiments. *p
Techniques Used: Western Blot, TUNEL Assay, Staining

Figure Legend Snippet: Depletion of PHD3 inhibits cardiomyocyte apoptosis induced by I/R injury After 5 doses of tamoxifen infusion, left anterior descending (LAD) coronary arteries of mice with indicated genotypes were tied for 40 minutes and then released for reperfusion. Twenty-four hours after reperfusion, hearts were fixed with 10% formaldehyde and embedded in paraffin. Cross sections of hearts were then analyzed with TUNEL staining. Nuclei were stained with DAPI. Representative high magnification images of the AAR are shown in (A) and low magnification images of whole sections are shown in (B) . Quantitative analysis of the apoptotic cells within the AAR is shown in (C). The numbers of mice analyzed are indicated in the bars respectively. *, p
Techniques Used: Mouse Assay, TUNEL Assay, Staining

Figure Legend Snippet: Depletion of PHD3 has no effect on HIF-1α protein level, the expression of HIF target genes or capillary density in the heart After 5 doses of tamoxifen, ventricles were excised and flash frozen. (A) Proteins extracted from ventricles of the indicated genotypes (n=3) were western-blotted with anti-HIF-1α and anti-PHD3 antibodies. Lysate from cardiomyocytes treated with DMOG (1mM) was used as the positive control for HIF-1α. (B) mRNAs were extracted from ventricles of the indicated genotypes. Relative mRNA level of HIF target genes and PHD3 were analyzed by quantitative real-time PCR. n = 3. (C) Heart sections of the indicated genotypes were immunostained with anti-myosin antibody and TRITC-E-lectin. Capillary densities are expressed as the number of lectin-positive objects per field of view. N.S., not significant, n = 3.
Techniques Used: Expressing, Western Blot, Positive Control, Real-time Polymerase Chain Reaction

Figure Legend Snippet: Depletion of PHD3 further stabilizes HIF-1α and overexpression of normoxia-stable HIF-1α protects cardiomyocytes from hypoxia-induced apoptosis (A) Neonatal ventricular myocytes from PHD3 f / f ; Cre +/− or PHD3 f / f ; Cre −/− mice were treated with 4-hydroxyl-tamoxifen for 3 days to delete PHD3. Cells were then cultured at 0.5% or 21% O 2 condition for 8 hours. Cells were then harvested for western blots with the indicated antibodies. (B), (C) HL-1 cardiomyocytes were infected with adenovirus expressing normoxia-stable HIF-1α-GFP (HIF-1α PP/AG -GFP) or lacZ for 24 hours. Infected cells were then cultured with fresh serum-free medium at 0.5% or 21% O 2 conditions for additional 48 hours to induce apoptosis. Cardiomyocytes were then fixed and stained with DAPI. Apoptosis was then analyzed with TUNEL staining. *, p
Techniques Used: Over Expression, Mouse Assay, Cell Culture, Western Blot, Infection, Expressing, Staining, TUNEL Assay

Figure Legend Snippet: DMOG inhibits HL-1 cardiomyocyte apoptosis induced by doxorubicin (A) HL-1 cells were transfected with two sets of si-RNA for Chk1 or scramble si-RNA as the control (Si-C) for two days. Cells were then treated with doxorubicin (1μM) as indicated. Western blots were then performed with the indicated antibodies. (B) After two days transfection with si-RNAs, HL-1 cells were treated with doxorubicin (1μM) for 8 hours and then harvested for caspase3/7 activity assay. Knocking down the expression of Chk-1 significantly inhibits caspase3/7 activity. N = 3, *p
Techniques Used: Transfection, Western Blot, Activity Assay, Expressing
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