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Gradient sds page gel electrophoresis

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4–15% gradient SDS-PAGE gel electrophoresis is a laboratory instrument used for separating and analyzing proteins based on their molecular weight. It utilizes a polyacrylamide gel with a gradient concentration of 4-15% to achieve effective separation of a wide range of protein sizes.

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Lab products found in correlation

Hrp conjugated goat anti rabbit, by Jackson ImmunoResearch (1 mentions) Total eif2α, by Cell Signaling Technology (1 mentions) Cleaved parp 1, by Cell Signaling Technology (1 mentions) Hrp conjugated goat anti rabbit or goat anti mouse, by Jackson ImmunoResearch (1 mentions) Mitochondria cytosol fractionation kit, by Abcam (1 mentions) Image studio lite quantification software, by LI COR (1 mentions) Xbp1s, by Cell Signaling Technology (1 mentions) P eif2α, by Cell Signaling Technology (1 mentions)

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SDS-PAGE (902 citations) SDS-PAGE gels (753 citations) 4–20% gradient gels (350 citations) 4–20% gradient SDS-PAGE (55 citations) 10% SDS gel (31 citations) SDS-PAGE electrophoresis (13 citations) 4–20% gradient SDS gel (6 citations) 4–15% gradient gel electrophoresis (3 citations) PAGE gels (3 citations)

2 protocols using gradient sds page gel electrophoresis

1

Protein Expression Analysis by Immunoblotting

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Cellular protein extraction, 4–15% gradient SDS-PAGE gel electrophoresis (Bio-Rad laboratories, Irvine, CA, USA), transfer to PVDF membrane, and immunoblot development were performed as published recently [37 (link)]. The following rabbit anti-human antibodies were used (obtained from Cell Signaling, Danvers, MA): cleaved PARP (5625), p-ERK (4370), ERK (4695), p-AKT (4060), AKT (4691), p-eIF2α (3398), eIF2α (5324), p-SAPK/JNK (4668), and SAPK/JNK (9252). Equal protein loading was examined by β-actin detection using a mouse monoclonal antibody (Sigma Aldrich); secondary antibodies: HRP-conjugated goat anti-rabbit or goat anti-mouse (Jackson ImmunoResearch Laboratories, West Grove, PA, USA). Densitometric image analysis was performed using Image StudioTM Lite quantification software (LI-COR Biosciences, Lincoln, NE, USA).
Jandova J., Hua A.B., Fimbres J, & Wondrak G.T. (2021). Deuterium Oxide (D2O) Induces Early Stress Response Gene Expression and Impairs Growth and Metastasis of Experimental Malignant Melanoma. Cancers, 13(4), 605.
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2

Cellular Protein Extraction and Analysis

Cited 1 time
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Cellular protein extraction, 4–15% gradient SDS-PAGE gel electrophoresis (Bio-Rad laboratories), transfer to PVDF membrane, and immunoblot development were performed as published recently.3 , 33 (link) The following rabbit anti-human antibodies were used (obtained from Cell Signaling): cleaved PARP-1 (5625), p-eIF2α (3398), total eIF2α (5324), SQSTM1 (8025), LAMP1 (9091), LC3-I/II (12741), cytochrome c (11940), ATF-4 (11815), and XBP-1s (40435). Equal protein loading was examined by β-actin detection using a mouse monoclonal antibody (Sigma Aldrich); secondary antibodies: HRP-conjugated goat anti-rabbit or goat anti-mouse (Jackson ImmunoResearch Laboratories). Densitometric image analysis was performed using Image Studio™ Lite quantification software (LI-COR Biosciences). For cytochrome c immunodetection, cell fractionation (mitochondrial versus cytosolic) was performed using the Mitochondria/Cytosol fractionation kit (ab65320, Abcam). Purity of fractions was confirmed according to kit instructions.
Jandova J., Park S.L., Corenblum M.J., Madhavan L., Snell J.A., Rounds L, & Wondrak G.T. (2022). Mefloquine induces ER stress and apoptosis in BRAFi-resistant A375-BRAFV600E/NRASQ61K malignant melanoma cells targeting intracranial tumors in a bioluminescent murine model. Molecular carcinogenesis, 61(6), 603-614.
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