Cells were grown on a glass coverslip for immunostaining and then provided with hypoxic treatment or the addition of CPO microparticles, as described in the cell culture section. Paraformaldehyde (PFA) (Sigma Aldrich) 4% in PBS for 15 mins was used to fix the cells, followed by permeabilization with 0.05% (v/v) Triton X-100 and blocking with 1% (w/v) BSA (Thermo Scientific) in PBS for 30 mins, all at RT. Next, fixed cells were stained with primary antibodies: Cardiac Troponin T Monoclonal Antibody (13-11), Mouse / IgG1 (Thermo Scientific), and hypoxia-inducible factor 1 (HIF-1α) antibody (H1Alpha67) (Novus Biologicals) for 1 hr. at RT. Following PBS washes, samples were incubated with phalloidin, DAPI, and goat anti-mouse secondary antibodies conjugated (Thermo Scientific) with either Alexa 594 or Alexa 488 (as applicable) for HIF-1α or cTn for 1 hr at RT. After washing with PBS, coverslips were mounted, and fluorescent images were acquired using a Keyence microscope with complementary Image software.
Hif 1α antibody h1alpha67
Manufactured by Novus Biologicals
The HIF-1α antibody (H1Alpha67) is a monoclonal antibody that binds to the hypoxia-inducible factor 1-alpha (HIF-1α) protein. HIF-1α is a subunit of the HIF-1 transcription factor, which plays a crucial role in the cellular response to hypoxia.
Automatically generated - may contain errors
2 protocols using hif 1α antibody h1alpha67
1
Immunofluorescence Analysis of Hypoxia-Induced Cells
2
Hypoxia and CPO Particle Effects on Cardiomyocyte Phenotype
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Cells were grown on a glass coverslip for immunostaining and then provided with hypoxic treatment or the addition of CPO microparticles, as described in the cell culture section. Paraformaldehyde (PFA) (Sigma Aldrich) 4% in PBS for 15 mins was used to fix the cells, followed by permeabilization with 0.05% (v/v) Triton X-100 and blocking with 1% (w/v) BSA (Thermo Scientific) in PBS for 30 mins, all at RT. Next, fixed cells were stained with primary antibodies: Cardiac Troponin T Monoclonal Antibody , Mouse / IgG1 (Thermo Scientific), and hypoxia-inducible factor 1 (HIF-1α) antibody (H1Alpha67) (Novus Biologicals) for 1 hr. at RT. Following PBS washes, samples were incubated with phalloidin, DAPI, and goat anti-mouse secondary antibodies conjugated (Thermo Scientific) with either Alexa 594 or Alexa 488 (as applicable) for HIF-1α or cTn for 1 hr at RT. After washing with PBS, coverslips were mounted, and fluorescent images were acquired using a Keyence microscope with complementary Image software.
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