p 50atf6α  (Abcam)

 
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    Anti ATF6 antibody
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    Abcam p 50atf6α
    SW induces ROS and ER stress, which interact with each other. (A-E) Effects of NAC on SW-induced ER stress. Following pretreatment with 5 mM NAC for 2 h, cells were treated with 25% SW for 4 h; then, the expression of the ER stress-associated proteins GRP78, <t>p-50ATF6α,</t> p-IRE1α, p-PERK and CHOP was evaluated by western blot analysis. (F) Effects of 4-PBA or Thap on SW-induced ROS generation. Following pretreatment with 2 mM 4-PBA or 150 nM Thap for 2 h, cells were treated with 25% SW for 4 h, and cellular ROS levels were assessed using 2′,7′-dichlorofluorescein diacetate and visualized with a fluorescence microscope (magnification, ×100). (G) Mean fluorescence intensity of (F) was analyzed using ImageJ software. (a) Control, (b) SW, (c) 4-PBA and (d) 4-PBA + SW groups. Data are presented as the means ± standard error of the mean, n=5. ** P

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    1) Product Images from "Seawater inhalation induces acute lung injury via ROS generation and the endoplasmic reticulum stress pathway"

    Article Title: Seawater inhalation induces acute lung injury via ROS generation and the endoplasmic reticulum stress pathway

    Journal: International Journal of Molecular Medicine

    doi: 10.3892/ijmm.2018.3486

    SW induces ROS and ER stress, which interact with each other. (A-E) Effects of NAC on SW-induced ER stress. Following pretreatment with 5 mM NAC for 2 h, cells were treated with 25% SW for 4 h; then, the expression of the ER stress-associated proteins GRP78, p-50ATF6α, p-IRE1α, p-PERK and CHOP was evaluated by western blot analysis. (F) Effects of 4-PBA or Thap on SW-induced ROS generation. Following pretreatment with 2 mM 4-PBA or 150 nM Thap for 2 h, cells were treated with 25% SW for 4 h, and cellular ROS levels were assessed using 2′,7′-dichlorofluorescein diacetate and visualized with a fluorescence microscope (magnification, ×100). (G) Mean fluorescence intensity of (F) was analyzed using ImageJ software. (a) Control, (b) SW, (c) 4-PBA and (d) 4-PBA + SW groups. Data are presented as the means ± standard error of the mean, n=5. ** P
    Figure Legend Snippet: SW induces ROS and ER stress, which interact with each other. (A-E) Effects of NAC on SW-induced ER stress. Following pretreatment with 5 mM NAC for 2 h, cells were treated with 25% SW for 4 h; then, the expression of the ER stress-associated proteins GRP78, p-50ATF6α, p-IRE1α, p-PERK and CHOP was evaluated by western blot analysis. (F) Effects of 4-PBA or Thap on SW-induced ROS generation. Following pretreatment with 2 mM 4-PBA or 150 nM Thap for 2 h, cells were treated with 25% SW for 4 h, and cellular ROS levels were assessed using 2′,7′-dichlorofluorescein diacetate and visualized with a fluorescence microscope (magnification, ×100). (G) Mean fluorescence intensity of (F) was analyzed using ImageJ software. (a) Control, (b) SW, (c) 4-PBA and (d) 4-PBA + SW groups. Data are presented as the means ± standard error of the mean, n=5. ** P

    Techniques Used: Expressing, Western Blot, Fluorescence, Microscopy, Software

    2) Product Images from "Seawater inhalation induces acute lung injury via ROS generation and the endoplasmic reticulum stress pathway"

    Article Title: Seawater inhalation induces acute lung injury via ROS generation and the endoplasmic reticulum stress pathway

    Journal: International Journal of Molecular Medicine

    doi: 10.3892/ijmm.2018.3486

    SW-induced cell apoptosis and growth inhibition are dependent on endoplasmic reticulum stress. (A) Effects of 4-PBA (30 mg/kg) on SW-induced apoptosis in rats (TUNEL staining). (a) Control, (b) SW, (c) 4-PBA and (d) 4-PBA + SW groups (magnification, ×10). (B) Effects of 4-PBA (2 mM) or Thap (150 nM) on SW-induced apoptosis of A549 cells (Annexin V-FITC/PI staining). (a) Control, (b) SW, (c) 4-PBA, (d) 4-PBA + SW and (e) Thap groups. (C) Number of apoptotic cells per field in (A). (D) Percentage of apoptotic cells in (B). (E-G) Effects of 4-PBA or Thap on the expression of apoptosis-associated proteins caspase-3 and p-JNK. (H) Effects of 4-PBA or Thap on SW-induced cell growth inhibition (Cell Counting kit-8 assay). Data are presented as the means ± standard error of the mean, n=5. * P
    Figure Legend Snippet: SW-induced cell apoptosis and growth inhibition are dependent on endoplasmic reticulum stress. (A) Effects of 4-PBA (30 mg/kg) on SW-induced apoptosis in rats (TUNEL staining). (a) Control, (b) SW, (c) 4-PBA and (d) 4-PBA + SW groups (magnification, ×10). (B) Effects of 4-PBA (2 mM) or Thap (150 nM) on SW-induced apoptosis of A549 cells (Annexin V-FITC/PI staining). (a) Control, (b) SW, (c) 4-PBA, (d) 4-PBA + SW and (e) Thap groups. (C) Number of apoptotic cells per field in (A). (D) Percentage of apoptotic cells in (B). (E-G) Effects of 4-PBA or Thap on the expression of apoptosis-associated proteins caspase-3 and p-JNK. (H) Effects of 4-PBA or Thap on SW-induced cell growth inhibition (Cell Counting kit-8 assay). Data are presented as the means ± standard error of the mean, n=5. * P

    Techniques Used: Inhibition, TUNEL Assay, Staining, Expressing, Cell Counting

    SW-induced growth inhibition and apoptosis depend on ROS. (A) Effects of NAC (150 mg/kg) pretreatment on SW-induced apoptosis in rats (TUNEL staining; magnification, ×10). (B) Effects of NAC (5 mM) on SW-induced apoptosis of A549 cells (Annexin V-FITC/PI staining). (a) Control, (b) SW, (c) NAC and (d) NAC + SW groups. (C) Number of apoptotic cells per field in (A). (D) Percentage of apoptotic cells in (B). (E-G) Effects of NAC (5 mM) on the expression of the apoptosis-associated proteins caspase-3 and p-JNK in A549 cells. (H) Effects of NAC on SW-induced cell growth inhibition (Cell Counting kit-8 assay). Data are presented as the means ± standard error of the mean, n=5. *** P
    Figure Legend Snippet: SW-induced growth inhibition and apoptosis depend on ROS. (A) Effects of NAC (150 mg/kg) pretreatment on SW-induced apoptosis in rats (TUNEL staining; magnification, ×10). (B) Effects of NAC (5 mM) on SW-induced apoptosis of A549 cells (Annexin V-FITC/PI staining). (a) Control, (b) SW, (c) NAC and (d) NAC + SW groups. (C) Number of apoptotic cells per field in (A). (D) Percentage of apoptotic cells in (B). (E-G) Effects of NAC (5 mM) on the expression of the apoptosis-associated proteins caspase-3 and p-JNK in A549 cells. (H) Effects of NAC on SW-induced cell growth inhibition (Cell Counting kit-8 assay). Data are presented as the means ± standard error of the mean, n=5. *** P

    Techniques Used: Inhibition, TUNEL Assay, Staining, Expressing, Cell Counting

    3) Product Images from "Seawater inhalation induces acute lung injury via ROS generation and the endoplasmic reticulum stress pathway"

    Article Title: Seawater inhalation induces acute lung injury via ROS generation and the endoplasmic reticulum stress pathway

    Journal: International Journal of Molecular Medicine

    doi: 10.3892/ijmm.2018.3486

    SW-induced cell apoptosis and growth inhibition are dependent on endoplasmic reticulum stress. (A) Effects of 4-PBA (30 mg/kg) on SW-induced apoptosis in rats (TUNEL staining). (a) Control, (b) SW, (c) 4-PBA and (d) 4-PBA + SW groups (magnification, ×10). (B) Effects of 4-PBA (2 mM) or Thap (150 nM) on SW-induced apoptosis of A549 cells (Annexin V-FITC/PI staining). (a) Control, (b) SW, (c) 4-PBA, (d) 4-PBA + SW and (e) Thap groups. (C) Number of apoptotic cells per field in (A). (D) Percentage of apoptotic cells in (B). (E-G) Effects of 4-PBA or Thap on the expression of apoptosis-associated proteins caspase-3 and p-JNK. (H) Effects of 4-PBA or Thap on SW-induced cell growth inhibition (Cell Counting kit-8 assay). Data are presented as the means ± standard error of the mean, n=5. * P
    Figure Legend Snippet: SW-induced cell apoptosis and growth inhibition are dependent on endoplasmic reticulum stress. (A) Effects of 4-PBA (30 mg/kg) on SW-induced apoptosis in rats (TUNEL staining). (a) Control, (b) SW, (c) 4-PBA and (d) 4-PBA + SW groups (magnification, ×10). (B) Effects of 4-PBA (2 mM) or Thap (150 nM) on SW-induced apoptosis of A549 cells (Annexin V-FITC/PI staining). (a) Control, (b) SW, (c) 4-PBA, (d) 4-PBA + SW and (e) Thap groups. (C) Number of apoptotic cells per field in (A). (D) Percentage of apoptotic cells in (B). (E-G) Effects of 4-PBA or Thap on the expression of apoptosis-associated proteins caspase-3 and p-JNK. (H) Effects of 4-PBA or Thap on SW-induced cell growth inhibition (Cell Counting kit-8 assay). Data are presented as the means ± standard error of the mean, n=5. * P

    Techniques Used: Inhibition, TUNEL Assay, Staining, Expressing, Cell Counting

    SW-induced growth inhibition and apoptosis depend on ROS. (A) Effects of NAC (150 mg/kg) pretreatment on SW-induced apoptosis in rats (TUNEL staining; magnification, ×10). (B) Effects of NAC (5 mM) on SW-induced apoptosis of A549 cells (Annexin V-FITC/PI staining). (a) Control, (b) SW, (c) NAC and (d) NAC + SW groups. (C) Number of apoptotic cells per field in (A). (D) Percentage of apoptotic cells in (B). (E-G) Effects of NAC (5 mM) on the expression of the apoptosis-associated proteins caspase-3 and p-JNK in A549 cells. (H) Effects of NAC on SW-induced cell growth inhibition (Cell Counting kit-8 assay). Data are presented as the means ± standard error of the mean, n=5. *** P
    Figure Legend Snippet: SW-induced growth inhibition and apoptosis depend on ROS. (A) Effects of NAC (150 mg/kg) pretreatment on SW-induced apoptosis in rats (TUNEL staining; magnification, ×10). (B) Effects of NAC (5 mM) on SW-induced apoptosis of A549 cells (Annexin V-FITC/PI staining). (a) Control, (b) SW, (c) NAC and (d) NAC + SW groups. (C) Number of apoptotic cells per field in (A). (D) Percentage of apoptotic cells in (B). (E-G) Effects of NAC (5 mM) on the expression of the apoptosis-associated proteins caspase-3 and p-JNK in A549 cells. (H) Effects of NAC on SW-induced cell growth inhibition (Cell Counting kit-8 assay). Data are presented as the means ± standard error of the mean, n=5. *** P

    Techniques Used: Inhibition, TUNEL Assay, Staining, Expressing, Cell Counting

    4) Product Images from "Seawater inhalation induces acute lung injury via ROS generation and the endoplasmic reticulum stress pathway"

    Article Title: Seawater inhalation induces acute lung injury via ROS generation and the endoplasmic reticulum stress pathway

    Journal: International Journal of Molecular Medicine

    doi: 10.3892/ijmm.2018.3486

    SW induces ROS and ER stress, which interact with each other. (A-E) Effects of NAC on SW-induced ER stress. Following pretreatment with 5 mM NAC for 2 h, cells were treated with 25% SW for 4 h; then, the expression of the ER stress-associated proteins GRP78, p-50ATF6α, p-IRE1α, p-PERK and CHOP was evaluated by western blot analysis. (F) Effects of 4-PBA or Thap on SW-induced ROS generation. Following pretreatment with 2 mM 4-PBA or 150 nM Thap for 2 h, cells were treated with 25% SW for 4 h, and cellular ROS levels were assessed using 2′,7′-dichlorofluorescein diacetate and visualized with a fluorescence microscope (magnification, ×100). (G) Mean fluorescence intensity of (F) was analyzed using ImageJ software. (a) Control, (b) SW, (c) 4-PBA and (d) 4-PBA + SW groups. Data are presented as the means ± standard error of the mean, n=5. ** P
    Figure Legend Snippet: SW induces ROS and ER stress, which interact with each other. (A-E) Effects of NAC on SW-induced ER stress. Following pretreatment with 5 mM NAC for 2 h, cells were treated with 25% SW for 4 h; then, the expression of the ER stress-associated proteins GRP78, p-50ATF6α, p-IRE1α, p-PERK and CHOP was evaluated by western blot analysis. (F) Effects of 4-PBA or Thap on SW-induced ROS generation. Following pretreatment with 2 mM 4-PBA or 150 nM Thap for 2 h, cells were treated with 25% SW for 4 h, and cellular ROS levels were assessed using 2′,7′-dichlorofluorescein diacetate and visualized with a fluorescence microscope (magnification, ×100). (G) Mean fluorescence intensity of (F) was analyzed using ImageJ software. (a) Control, (b) SW, (c) 4-PBA and (d) 4-PBA + SW groups. Data are presented as the means ± standard error of the mean, n=5. ** P

    Techniques Used: Expressing, Western Blot, Fluorescence, Microscopy, Software

    5) Product Images from "Seawater inhalation induces acute lung injury via ROS generation and the endoplasmic reticulum stress pathway"

    Article Title: Seawater inhalation induces acute lung injury via ROS generation and the endoplasmic reticulum stress pathway

    Journal: International Journal of Molecular Medicine

    doi: 10.3892/ijmm.2018.3486

    SW induces ROS and ER stress, which interact with each other. (A-E) Effects of NAC on SW-induced ER stress. Following pretreatment with 5 mM NAC for 2 h, cells were treated with 25% SW for 4 h; then, the expression of the ER stress-associated proteins GRP78, p-50ATF6α, p-IRE1α, p-PERK and CHOP was evaluated by western blot analysis. (F) Effects of 4-PBA or Thap on SW-induced ROS generation. Following pretreatment with 2 mM 4-PBA or 150 nM Thap for 2 h, cells were treated with 25% SW for 4 h, and cellular ROS levels were assessed using 2′,7′-dichlorofluorescein diacetate and visualized with a fluorescence microscope (magnification, ×100). (G) Mean fluorescence intensity of (F) was analyzed using ImageJ software. (a) Control, (b) SW, (c) 4-PBA and (d) 4-PBA + SW groups. Data are presented as the means ± standard error of the mean, n=5. ** P
    Figure Legend Snippet: SW induces ROS and ER stress, which interact with each other. (A-E) Effects of NAC on SW-induced ER stress. Following pretreatment with 5 mM NAC for 2 h, cells were treated with 25% SW for 4 h; then, the expression of the ER stress-associated proteins GRP78, p-50ATF6α, p-IRE1α, p-PERK and CHOP was evaluated by western blot analysis. (F) Effects of 4-PBA or Thap on SW-induced ROS generation. Following pretreatment with 2 mM 4-PBA or 150 nM Thap for 2 h, cells were treated with 25% SW for 4 h, and cellular ROS levels were assessed using 2′,7′-dichlorofluorescein diacetate and visualized with a fluorescence microscope (magnification, ×100). (G) Mean fluorescence intensity of (F) was analyzed using ImageJ software. (a) Control, (b) SW, (c) 4-PBA and (d) 4-PBA + SW groups. Data are presented as the means ± standard error of the mean, n=5. ** P

    Techniques Used: Expressing, Western Blot, Fluorescence, Microscopy, Software

    6) Product Images from "Seawater inhalation induces acute lung injury via ROS generation and the endoplasmic reticulum stress pathway"

    Article Title: Seawater inhalation induces acute lung injury via ROS generation and the endoplasmic reticulum stress pathway

    Journal: International Journal of Molecular Medicine

    doi: 10.3892/ijmm.2018.3486

    SW induces ROS and ER stress, which interact with each other. (A-E) Effects of NAC on SW-induced ER stress. Following pretreatment with 5 mM NAC for 2 h, cells were treated with 25% SW for 4 h; then, the expression of the ER stress-associated proteins GRP78, p-50ATF6α, p-IRE1α, p-PERK and CHOP was evaluated by western blot analysis. (F) Effects of 4-PBA or Thap on SW-induced ROS generation. Following pretreatment with 2 mM 4-PBA or 150 nM Thap for 2 h, cells were treated with 25% SW for 4 h, and cellular ROS levels were assessed using 2′,7′-dichlorofluorescein diacetate and visualized with a fluorescence microscope (magnification, ×100). (G) Mean fluorescence intensity of (F) was analyzed using ImageJ software. (a) Control, (b) SW, (c) 4-PBA and (d) 4-PBA + SW groups. Data are presented as the means ± standard error of the mean, n=5. ** P
    Figure Legend Snippet: SW induces ROS and ER stress, which interact with each other. (A-E) Effects of NAC on SW-induced ER stress. Following pretreatment with 5 mM NAC for 2 h, cells were treated with 25% SW for 4 h; then, the expression of the ER stress-associated proteins GRP78, p-50ATF6α, p-IRE1α, p-PERK and CHOP was evaluated by western blot analysis. (F) Effects of 4-PBA or Thap on SW-induced ROS generation. Following pretreatment with 2 mM 4-PBA or 150 nM Thap for 2 h, cells were treated with 25% SW for 4 h, and cellular ROS levels were assessed using 2′,7′-dichlorofluorescein diacetate and visualized with a fluorescence microscope (magnification, ×100). (G) Mean fluorescence intensity of (F) was analyzed using ImageJ software. (a) Control, (b) SW, (c) 4-PBA and (d) 4-PBA + SW groups. Data are presented as the means ± standard error of the mean, n=5. ** P

    Techniques Used: Expressing, Western Blot, Fluorescence, Microscopy, Software

    7) Product Images from "Seawater inhalation induces acute lung injury via ROS generation and the endoplasmic reticulum stress pathway"

    Article Title: Seawater inhalation induces acute lung injury via ROS generation and the endoplasmic reticulum stress pathway

    Journal: International Journal of Molecular Medicine

    doi: 10.3892/ijmm.2018.3486

    SW-induced cell apoptosis and growth inhibition are dependent on endoplasmic reticulum stress. (A) Effects of 4-PBA (30 mg/kg) on SW-induced apoptosis in rats (TUNEL staining). (a) Control, (b) SW, (c) 4-PBA and (d) 4-PBA + SW groups (magnification, ×10). (B) Effects of 4-PBA (2 mM) or Thap (150 nM) on SW-induced apoptosis of A549 cells (Annexin V-FITC/PI staining). (a) Control, (b) SW, (c) 4-PBA, (d) 4-PBA + SW and (e) Thap groups. (C) Number of apoptotic cells per field in (A). (D) Percentage of apoptotic cells in (B). (E-G) Effects of 4-PBA or Thap on the expression of apoptosis-associated proteins caspase-3 and p-JNK. (H) Effects of 4-PBA or Thap on SW-induced cell growth inhibition (Cell Counting kit-8 assay). Data are presented as the means ± standard error of the mean, n=5. * P
    Figure Legend Snippet: SW-induced cell apoptosis and growth inhibition are dependent on endoplasmic reticulum stress. (A) Effects of 4-PBA (30 mg/kg) on SW-induced apoptosis in rats (TUNEL staining). (a) Control, (b) SW, (c) 4-PBA and (d) 4-PBA + SW groups (magnification, ×10). (B) Effects of 4-PBA (2 mM) or Thap (150 nM) on SW-induced apoptosis of A549 cells (Annexin V-FITC/PI staining). (a) Control, (b) SW, (c) 4-PBA, (d) 4-PBA + SW and (e) Thap groups. (C) Number of apoptotic cells per field in (A). (D) Percentage of apoptotic cells in (B). (E-G) Effects of 4-PBA or Thap on the expression of apoptosis-associated proteins caspase-3 and p-JNK. (H) Effects of 4-PBA or Thap on SW-induced cell growth inhibition (Cell Counting kit-8 assay). Data are presented as the means ± standard error of the mean, n=5. * P

    Techniques Used: Inhibition, TUNEL Assay, Staining, Expressing, Cell Counting

    SW-induced growth inhibition and apoptosis depend on ROS. (A) Effects of NAC (150 mg/kg) pretreatment on SW-induced apoptosis in rats (TUNEL staining; magnification, ×10). (B) Effects of NAC (5 mM) on SW-induced apoptosis of A549 cells (Annexin V-FITC/PI staining). (a) Control, (b) SW, (c) NAC and (d) NAC + SW groups. (C) Number of apoptotic cells per field in (A). (D) Percentage of apoptotic cells in (B). (E-G) Effects of NAC (5 mM) on the expression of the apoptosis-associated proteins caspase-3 and p-JNK in A549 cells. (H) Effects of NAC on SW-induced cell growth inhibition (Cell Counting kit-8 assay). Data are presented as the means ± standard error of the mean, n=5. *** P
    Figure Legend Snippet: SW-induced growth inhibition and apoptosis depend on ROS. (A) Effects of NAC (150 mg/kg) pretreatment on SW-induced apoptosis in rats (TUNEL staining; magnification, ×10). (B) Effects of NAC (5 mM) on SW-induced apoptosis of A549 cells (Annexin V-FITC/PI staining). (a) Control, (b) SW, (c) NAC and (d) NAC + SW groups. (C) Number of apoptotic cells per field in (A). (D) Percentage of apoptotic cells in (B). (E-G) Effects of NAC (5 mM) on the expression of the apoptosis-associated proteins caspase-3 and p-JNK in A549 cells. (H) Effects of NAC on SW-induced cell growth inhibition (Cell Counting kit-8 assay). Data are presented as the means ± standard error of the mean, n=5. *** P

    Techniques Used: Inhibition, TUNEL Assay, Staining, Expressing, Cell Counting

    8) Product Images from "Seawater inhalation induces acute lung injury via ROS generation and the endoplasmic reticulum stress pathway"

    Article Title: Seawater inhalation induces acute lung injury via ROS generation and the endoplasmic reticulum stress pathway

    Journal: International Journal of Molecular Medicine

    doi: 10.3892/ijmm.2018.3486

    SW induces ROS and ER stress, which interact with each other. (A-E) Effects of NAC on SW-induced ER stress. Following pretreatment with 5 mM NAC for 2 h, cells were treated with 25% SW for 4 h; then, the expression of the ER stress-associated proteins GRP78, p-50ATF6α, p-IRE1α, p-PERK and CHOP was evaluated by western blot analysis. (F) Effects of 4-PBA or Thap on SW-induced ROS generation. Following pretreatment with 2 mM 4-PBA or 150 nM Thap for 2 h, cells were treated with 25% SW for 4 h, and cellular ROS levels were assessed using 2′,7′-dichlorofluorescein diacetate and visualized with a fluorescence microscope (magnification, ×100). (G) Mean fluorescence intensity of (F) was analyzed using ImageJ software. (a) Control, (b) SW, (c) 4-PBA and (d) 4-PBA + SW groups. Data are presented as the means ± standard error of the mean, n=5. ** P
    Figure Legend Snippet: SW induces ROS and ER stress, which interact with each other. (A-E) Effects of NAC on SW-induced ER stress. Following pretreatment with 5 mM NAC for 2 h, cells were treated with 25% SW for 4 h; then, the expression of the ER stress-associated proteins GRP78, p-50ATF6α, p-IRE1α, p-PERK and CHOP was evaluated by western blot analysis. (F) Effects of 4-PBA or Thap on SW-induced ROS generation. Following pretreatment with 2 mM 4-PBA or 150 nM Thap for 2 h, cells were treated with 25% SW for 4 h, and cellular ROS levels were assessed using 2′,7′-dichlorofluorescein diacetate and visualized with a fluorescence microscope (magnification, ×100). (G) Mean fluorescence intensity of (F) was analyzed using ImageJ software. (a) Control, (b) SW, (c) 4-PBA and (d) 4-PBA + SW groups. Data are presented as the means ± standard error of the mean, n=5. ** P

    Techniques Used: Expressing, Western Blot, Fluorescence, Microscopy, Software

    9) Product Images from "Seawater inhalation induces acute lung injury via ROS generation and the endoplasmic reticulum stress pathway"

    Article Title: Seawater inhalation induces acute lung injury via ROS generation and the endoplasmic reticulum stress pathway

    Journal: International Journal of Molecular Medicine

    doi: 10.3892/ijmm.2018.3486

    SW induces ROS and ER stress, which interact with each other. (A-E) Effects of NAC on SW-induced ER stress. Following pretreatment with 5 mM NAC for 2 h, cells were treated with 25% SW for 4 h; then, the expression of the ER stress-associated proteins GRP78, p-50ATF6α, p-IRE1α, p-PERK and CHOP was evaluated by western blot analysis. (F) Effects of 4-PBA or Thap on SW-induced ROS generation. Following pretreatment with 2 mM 4-PBA or 150 nM Thap for 2 h, cells were treated with 25% SW for 4 h, and cellular ROS levels were assessed using 2′,7′-dichlorofluorescein diacetate and visualized with a fluorescence microscope (magnification, ×100). (G) Mean fluorescence intensity of (F) was analyzed using ImageJ software. (a) Control, (b) SW, (c) 4-PBA and (d) 4-PBA + SW groups. Data are presented as the means ± standard error of the mean, n=5. ** P
    Figure Legend Snippet: SW induces ROS and ER stress, which interact with each other. (A-E) Effects of NAC on SW-induced ER stress. Following pretreatment with 5 mM NAC for 2 h, cells were treated with 25% SW for 4 h; then, the expression of the ER stress-associated proteins GRP78, p-50ATF6α, p-IRE1α, p-PERK and CHOP was evaluated by western blot analysis. (F) Effects of 4-PBA or Thap on SW-induced ROS generation. Following pretreatment with 2 mM 4-PBA or 150 nM Thap for 2 h, cells were treated with 25% SW for 4 h, and cellular ROS levels were assessed using 2′,7′-dichlorofluorescein diacetate and visualized with a fluorescence microscope (magnification, ×100). (G) Mean fluorescence intensity of (F) was analyzed using ImageJ software. (a) Control, (b) SW, (c) 4-PBA and (d) 4-PBA + SW groups. Data are presented as the means ± standard error of the mean, n=5. ** P

    Techniques Used: Expressing, Western Blot, Fluorescence, Microscopy, Software

    10) Product Images from "Seawater inhalation induces acute lung injury via ROS generation and the endoplasmic reticulum stress pathway"

    Article Title: Seawater inhalation induces acute lung injury via ROS generation and the endoplasmic reticulum stress pathway

    Journal: International Journal of Molecular Medicine

    doi: 10.3892/ijmm.2018.3486

    SW-induced cell apoptosis and growth inhibition are dependent on endoplasmic reticulum stress. (A) Effects of 4-PBA (30 mg/kg) on SW-induced apoptosis in rats (TUNEL staining). (a) Control, (b) SW, (c) 4-PBA and (d) 4-PBA + SW groups (magnification, ×10). (B) Effects of 4-PBA (2 mM) or Thap (150 nM) on SW-induced apoptosis of A549 cells (Annexin V-FITC/PI staining). (a) Control, (b) SW, (c) 4-PBA, (d) 4-PBA + SW and (e) Thap groups. (C) Number of apoptotic cells per field in (A). (D) Percentage of apoptotic cells in (B). (E-G) Effects of 4-PBA or Thap on the expression of apoptosis-associated proteins caspase-3 and p-JNK. (H) Effects of 4-PBA or Thap on SW-induced cell growth inhibition (Cell Counting kit-8 assay). Data are presented as the means ± standard error of the mean, n=5. * P
    Figure Legend Snippet: SW-induced cell apoptosis and growth inhibition are dependent on endoplasmic reticulum stress. (A) Effects of 4-PBA (30 mg/kg) on SW-induced apoptosis in rats (TUNEL staining). (a) Control, (b) SW, (c) 4-PBA and (d) 4-PBA + SW groups (magnification, ×10). (B) Effects of 4-PBA (2 mM) or Thap (150 nM) on SW-induced apoptosis of A549 cells (Annexin V-FITC/PI staining). (a) Control, (b) SW, (c) 4-PBA, (d) 4-PBA + SW and (e) Thap groups. (C) Number of apoptotic cells per field in (A). (D) Percentage of apoptotic cells in (B). (E-G) Effects of 4-PBA or Thap on the expression of apoptosis-associated proteins caspase-3 and p-JNK. (H) Effects of 4-PBA or Thap on SW-induced cell growth inhibition (Cell Counting kit-8 assay). Data are presented as the means ± standard error of the mean, n=5. * P

    Techniques Used: Inhibition, TUNEL Assay, Staining, Expressing, Cell Counting

    SW-induced growth inhibition and apoptosis depend on ROS. (A) Effects of NAC (150 mg/kg) pretreatment on SW-induced apoptosis in rats (TUNEL staining; magnification, ×10). (B) Effects of NAC (5 mM) on SW-induced apoptosis of A549 cells (Annexin V-FITC/PI staining). (a) Control, (b) SW, (c) NAC and (d) NAC + SW groups. (C) Number of apoptotic cells per field in (A). (D) Percentage of apoptotic cells in (B). (E-G) Effects of NAC (5 mM) on the expression of the apoptosis-associated proteins caspase-3 and p-JNK in A549 cells. (H) Effects of NAC on SW-induced cell growth inhibition (Cell Counting kit-8 assay). Data are presented as the means ± standard error of the mean, n=5. *** P
    Figure Legend Snippet: SW-induced growth inhibition and apoptosis depend on ROS. (A) Effects of NAC (150 mg/kg) pretreatment on SW-induced apoptosis in rats (TUNEL staining; magnification, ×10). (B) Effects of NAC (5 mM) on SW-induced apoptosis of A549 cells (Annexin V-FITC/PI staining). (a) Control, (b) SW, (c) NAC and (d) NAC + SW groups. (C) Number of apoptotic cells per field in (A). (D) Percentage of apoptotic cells in (B). (E-G) Effects of NAC (5 mM) on the expression of the apoptosis-associated proteins caspase-3 and p-JNK in A549 cells. (H) Effects of NAC on SW-induced cell growth inhibition (Cell Counting kit-8 assay). Data are presented as the means ± standard error of the mean, n=5. *** P

    Techniques Used: Inhibition, TUNEL Assay, Staining, Expressing, Cell Counting

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    Incubation:

    Article Title: Endoplasmic Reticulum Stress in Spinal Cord Contributes to the Development of Morphine Tolerance
    Article Snippet: .. The sections were then incubated overnight at 4°C with the following primary antibodies: rabbit anti-BiP antibody (1:500; ab21685; Abcam, Cambridge, MA, United States), rabbit anti-IRE1 antibody (1:50; ab37073; Abcam, Cambridge, MA, United States), rabbit anti-XBP1 antibody (1:50; A1731; ABclonal, Wuhan, China), rabbit anti-PERK (phospho T981) antibody (1:50; YP1055; ImmunoWay, Plano, TX, United States), rabbit anti-phospho-eIF2α (Ser51) (D9G8) antibody (1:50; #3398; CST, Beverly, MA, United States), rabbit anti-ATF6 antibody (1:50; ab203119; Abcam, Cambridge, MA, United States). .. Then, the sections were incubated with Alexa Fluor 488-labeled donkey anti-rabbit secondary antibody (1:300; A-21206; Invitrogen, Carlsbad, CA, United States) for 2 h at RT and washed with PBS.

    Article Title: Titanium dioxide nanoparticles induce endoplasmic reticulum stress-mediated apoptotic cell death in liver cancer cells
    Article Snippet: .. The sections were then incubated with specific primary monoclonal antibodies against ATF6 (dilution 1:250; Abcam, Cambridge, MA, USA) and phospho-PERK (dilution 1:200; Abcam) for 12 hours at 4°C, followed by incubation with horseradish peroxidase (HRP)-labeled goat anti-rabbit/rat IgG secondary antibody (dilution 1:1000, Santa Cruz Biotechnology, Santa Cruz, CA, USA) at room temperature for 4 hours. .. ATF6- and PERK-positive cells were visualized using diaminobenzidine staining and images were obtained using an Olympus microscope (BX51T-PHD-J11, ×400, Olympus Corporation, Tokyo, Japan) after counterstaining with hematoxylin.

    Binding Assay:

    Article Title: Seawater inhalation induces acute lung injury via ROS generation and the endoplasmic reticulum stress pathway
    Article Snippet: .. Reagents Antibodies against phosphorylated (p)-protein kinase R-like ER kinase (PERK; ab192591), PERK (ab79483), inositol-requiring kinase 1α (IRE1α; ab37073), p-IRE1α (ab48187), activating transcription factor 6α (ATF6α), p-50ATF6α (ab37149), glucose-regulated protein 78 (GRP78; ab21685), CCAAT/enhancer binding protein homologous protein (CHOP; ab11419), p-c-Jun N-terminal kinase (JNK; ab124956), JNK (ab179461) and caspase-3 (ab13847) were purchased from Abcam (Cambridge, UK). .. N-acetyl L-cysteine (NAC), 4-phenylbutyric acid (4-PBA) and thap-sigargin (Thap) were also purchased from Abcam.

    other:

    Article Title: Matrine attenuates endoplasmic reticulum stress and mitochondrion dysfunction in nonalcoholic fatty liver disease by regulating SERCA pathway
    Article Snippet: Other antibodies included anti-ATF6 (ab37149), anti-phospho-IRE1 (phospho S724) (ab48187), anti-IRE1 (ab37073), anti-SREBP1 (ab28481), Anti-Fatty Acid Synthase (ab22759), anti-Acetyl Coenzyme A Carboxylase (ab45174), anti-NF-κB p65, (ab16502), anti-c-jun (phospho S63) (ab32385) were purchased from Abcam (Cambridge, UK).

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    Abcam thap sigargin thap
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