Western Blot Techniques

Kim, Brianna

doi:10.1007/978-1-4939-6990-6_9

Cited by 105 publications

(56 citation statements)

References 6 publications

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“…Equal amount of proteins were added into SDS-PAGE and transferred to PVDF membranes according to the previous reports. [21] The antibodies (USA Abcam company) used were as follows: CYP27A1 1:1000, CYP46A1 1:1000, CYP7B1 1:1000, CYP7A1 1:1000, Cathepsin B 1:1000, Cathepsin D 1:1000, and Goat anti-rabbit IgG 1:5000. Fluorchem FC 2 software was used to analyze the image and the relative expression of target proteins was calculated with the following formula: relative coefficient = target protein/GAPDH.…”

Section: Western Blotmentioning

confidence: 99%

Increased Levels of 27‐Hydroxycholesterol Induced by Dietary Cholesterol in Brain Contribute to Learning and Memory Impairment in Rats

Zhang

et al. 2018

Molecular Nutrition Food Res

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Section: Western Blotmentioning

confidence: 99%

Increased Levels of 27‐Hydroxycholesterol Induced by Dietary Cholesterol in Brain Contribute to Learning and Memory Impairment in Rats

Zhang

et al. 2018

Molecular Nutrition Food Res

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“…The tested strains K pneumoniae ZJ02, E coli DZ2‐12R or E coli BL21 (DE3) (pET28a‐ mcr‐1 ) were cocultured with various concentrations of IAL (0, 4 and 32 μg/mL) as described in the growth curve determination for 4 hours. Following centrifugation at 13 800 g for 5 min, the precipitate of each sample was resuspended in loading buffer, mixed, boiled at 100°C for 7 minutes and separated by SDS‐PAGE . Then, the protein was transferred to a PVDF membrane.…”

Section: Methodsmentioning

confidence: 99%

“…Following centrifugation at 13 800 g for 5 min, the precipitate of each sample was resuspended in loading buffer, mixed, boiled at 100°C for 7 minutes and separated by SDS-PAGE. 27 Then, the protein was transferred to a PVDF membrane. After blocking, the mouse monoclonal antibody against MCR-1 (1:2000, Laboratory preservation), goat antimouse HRP-conjugated secondary antibodies (1:2000, Proteintech) were applied for the determination of MCR-1 production as described in our previous study.…”

Section: Western Blot Assaymentioning

confidence: 99%

Isoalantolactone restores the sensitivity of gram‐negative Enterobacteriaceae carrying MCR‐1 to carbapenems

Lü

Sun

et al. 2020

J Cellular Molecular Medi

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Polymyxin B has been re‐applied to the clinic as the final choice for the treatment of multidrug‐resistant gram‐negative pathogenic infections, but the use of polymyxin B has been re‐assessed because of the emergence and spread of the plasmid‐mediated mcr‐1 gene. The purpose of this study was to search for an MCR inhibitor synergistically acting with polymyxin to treat the infection caused by this pathogen. In this study, we used the broth microdilution checkerboard method to evaluate the synergistic effect of isoalantolactone (IAL) and polymyxin B on mcr‐1‐positive Enterobacteriaceae. Growth curve analysis, time‐killing assays and a combined disc test were used to further verify the efficacy of the combined drug. Colonization of the thigh muscle in mice, survival experiments and lung tissue section observations was used to determine the effect of synergy in vivo after Klebsiella pneumoniae and Escherichia coli infection. We screened a natural compound, IAL, which can enhance the sensitivity of polymyxin B to mcr‐1‐positive Enterobacteriaceae. The results showed that the combined use of polymyxin B and IAL has a synergistic effect on mcr‐1‐positive Enterobacteriaceae, such as K pneumoniae and E coli, not only in vitro but also in vivo. Our results indicate that IAL is a natural compound with broad application prospects that can prolong the service life of polymyxin B and make outstanding contributions to the treatment of gram‐negative Enterobacteriaceae infections resistant to polymyxin B.

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“…(1) Western blot was performed to detect TRPC1, 3, and 6 transcription levels in the normal control group, hypertonic control group, and HG group after 24 and 48 h and in different concentrations of SKF96365 following HG treatment for 24 h and VEGF protein expression was also determined [20]. Radio immunoprecipitation assay Lysis Buffer was used to extract total protein, and total protein concentration in the samples was assessed using the BCA method.…”

Section: Effects Of Hg and Skf96365 On Trpc And Vegf Protein Expressimentioning

confidence: 99%

The Effect and Mechanism of TRPC1, 3, and 6 on the Proliferation, Migration, and Lumen Formation of Retinal Vascular Endothelial Cells Induced by High Glucose

et al. 2020

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Objective: Transient receptor potential canonical (TRPC) channels are involved in neovascularization repairing after vascular injury in many tissues. However, whether TRPCs play a regulatory role in the development of diabetic retinopathy (DR) has rarely been reported. In the present study, we selected TRPC1, 3, and 6 to determine their roles and mechanism in human retina vascular endothelial cells (HREC) under high glucose (HG) conditions. Methods: HRECs were cultured in vitro under HG, hyper osmosis, and normal conditions. The expression of TRPC1, 3, and 6 in the cells at 24 and 48 h were detected by RT-polymerase chain reaction (PCR), Western blot and cell immunohistochemistry (IHC); In various concentrations, SKF96365 acted on HG cultured HRECs, the expression of vascular endothelial growth factor (VEGF) were detected by the same methods above; and the CCK-8, Transwell, cell scratch assay, and Matrigel assay were used to assess cell proliferation, migration, and lumen formation. Results: The RT-PCR, Western blot, and IHC results showed that TRPC1 expression was increased, and TRPC6 mRNA expression was increased under high-glucose conditions. SKF96365 acted on HG cultured HRECs that VEGF expression was significantly decreased. The CCK-8 assay, Transwell assay, cell scratch assay, and Matrigel assay showed that cell proliferation, migration, and lumen formation were downregulated by SKF96365. Conclusion: HG can induce increased expression of TRPC1 and 6 in HRECs. Inhibition of the TRPC pathway not only can decrease VEGF expression but also can prevent proliferation, migration, and lumen formation of HRECs induced by HG. Inhibition of TRPC channels is expected to become a drug target for

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Western Blot Techniques

Cited by 105 publications

References 6 publications

Increased Levels of 27‐Hydroxycholesterol Induced by Dietary Cholesterol in Brain Contribute to Learning and Memory Impairment in Rats

Increased Levels of 27‐Hydroxycholesterol Induced by Dietary Cholesterol in Brain Contribute to Learning and Memory Impairment in Rats

Isoalantolactone restores the sensitivity of gram‐negative Enterobacteriaceae carrying MCR‐1 to carbapenems

The Effect and Mechanism of TRPC1, 3, and 6 on the Proliferation, Migration, and Lumen Formation of Retinal Vascular Endothelial Cells Induced by High Glucose

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