The oxidative stress in the brain at 24 hours after CA was evaluated by immunohistochemical staining of 8-hydroxy-2'-deoxyguanosine (8-OHdG). Brain tissues were sectioned and embedded in paraffin. The immunohistochemistry was performed as we previously described (28 (link)), and 8-OHdG antibody (diluted 1:200; Abcam, Cambridge, UK) was used. The immunostaining scores were estimated using both the percentage of positively stained cells and the staining intensity as we previously described (17 (link)). The ROS levels in the brain at 6 hours after CA were measured using the tissue ROS assay kit (Genmed Scientifics, Wilmington, DE, USA) that utilized 2’,7’-dichlorofluorescein diacetate as the oxidative fluorescent probe. After the fresh brain tissues were isolated, the ROS levels were measured following the manufacturer’s instructions.
8 ohdg antibody
Manufactured by Abcam
Sourced in United Kingdom
The 8-OHdG antibody is a tool used in research to detect and measure the levels of 8-hydroxy-2'-deoxyguanosine (8-OHdG), a biomarker for oxidative stress and DNA damage. The antibody specifically binds to 8-OHdG, allowing for the quantification of this molecule in various biological samples.
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Lab products found in correlation
Tissue ros assay kit, by Genmed Scientifics (1 mentions)
Calnexin antibody, by Cell Signaling Technology (1 mentions)
Pdi antibody, by Cell Signaling Technology (1 mentions)
Anti mouse lgg hrp linked antibody, by Cell Signaling Technology (1 mentions)
Anti rabbit lgg hrp linked antibody, by Cell Signaling Technology (1 mentions)
Jnk antibody, by Cell Signaling Technology (1 mentions)
Estradiol elisa kit, by Abcam (1 mentions)
β actin, by Cell Signaling Technology (1 mentions)
P jnk antibody, by Cell Signaling Technology (1 mentions)
Optical microscope, by Olympus (1 mentions)
Diethylenetriamine, by Merck Group (1 mentions)
Chloroauric acid, by Merck Group (1 mentions)
Methylbenzene, by Merck Group (1 mentions)
Dehydrated alcohol, by Merck Group (1 mentions)
1 dodecanethiol, by Merck Group (1 mentions)
11 mercaptoundecanoic acid, by Merck Group (1 mentions)
3 glycidyloxypropyl trimethoxysilane, by Merck Group (1 mentions)
3 mercaptopropionic acid, by Merck Group (1 mentions)
4 nitrophenol, by Merck Group (1 mentions)
8 ohdg, by Merck Group (1 mentions)
6 protocols using 8 ohdg antibody
1
Oxidative Stress Evaluation in Brain after Cardiac Arrest
2
Measuring Oxidative DNA Damage in MDA-MB-231 Cells
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The levels of 8-OH-dG were measured using immunofluorescence. MDA-MB-231 cells, treated with different doses of simvastatin, were fixed by 4% PFA for 30min, the culture was washed with PBS 3 times. After permeabilization, 50μL 8-OH-dG antibody (Abcam, ab48508) was added at 1:200 dilution 37°C for 30 min. After several washes, cells were incubated with Alexa Fluor 594 donkey anti-goat IgG secondary antibody (Life Technologies, cat#A-11058).
3
Estrogen-Induced Oxidative Stress Response
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OVX rat injected E2 (Tocris Bioscience, Bristol, UK; catalog no. 2824), Estradiol ELISA kit (Biovision, Milpitas, CA, USA; catalog no. K7417-100). The primary antibodies used for western blot and immunofluorescence staining were 8-OHDG antibody (Abcam, Cambridge, UK; catalog no. ab48508), BiP antibody (Thermo Fisher Scientific, Lafayette, Colorado; catalog no. PA1-014A), Sigma receptor 1 antibody (Santa Cruz, California, USA; catalog no. sc-137075), pJNK antibody (Cell Signaling Technology, Boston, Massachusetts, USA; catalog no. 4668), JNK antibody (Cell Signaling Technology; catalog no. 9252), PDI antibody (Cell Signaling Technology; catalog no.3501), Ero-1α antibody (Cell Signaling Technology; catalog no.3264), Calnexin antibody (Cell Signaling Technology; catalog no.2679) and β-actin (Cell Signaling Technology; catalog no. 8457). The secondary antibodies used for western blot and immunofluorescence staining were the anti-rabbit lgG, HRP linked antibody (Cell Signaling Technology; catalog no. 7074), anti-mouse lgG, HRP linked antibody (Cell Signaling Technology; catalog no. 7076), Goat anti-Mouse IgG (H + L) Highly Cross-Adsorbed Secondary Antibody, Alexa Fluor Plus 647 (Thermo Fisher Scientific; catalog no. A32728) and Goat anti-rabbit IgG (H + L) Highly Cross-Adsorbed Secondary Antibody, Alexa Fluor Plus 488 (Thermo Fisher Scientific; catalog no. A-11029).
4
Quantification of 8-OHdG in Tumor Tissues
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Mice were sacrificed when their tumors reached 10% of their body weight. Primary tumors and organs were harvested, embedded in paraffin, sectioned, and stained with 8-hydroxydeoxyguanosine (8-OHdG) antibody (Abcam, 1:200 dilution) overnight at 4°C. After being washed in PBS, slides were incubated for secondary antibody for 1 h and examined using an Olympus optical microscope. The immunohistochemical staining results in each group were quantified by three observers.
5
Nanoporous Sensor for 8-OHdG Detection
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Specifically, 8-OHdG, bovine serum albumin (BSA), citric acid, diethylenetriamine (EDTA), (3-glycidyloxypropyl)trimethoxysilane (GPMS), glutaraldehyde, methylbenzene, chloroauric acid, 3-mercaptopropionic acid (MPA), zinc nitrate hexahydrate (Zn(NO3)2·6H2O), methanol (99.8%), sodium chloride, potassium chloride, calcium chloride, acetone, N,N-dimethylformamide (DMF), sodium borohydride (NaBH4, 99.99%), and dehydrated alcohol were obtained from Sigma Aldrich (St. Louis, Missouri (Mo), USA). NaCl, KCl, CaCl2, MgCl2, thymine, cytosine, adenine, guanine, and hydrogen peroxide were ordered from ALADDIN Reagent (Shanghai, China). Nanoporous alumina membranes were purchased from Whatman (Boston, Massachusetts (Ma), USA). Alongside, 8-OHdG antibody was bought from Abcam (Cambridge, UK). In addition, 2-methylimidazole (2-MeIM, 99%), 1-dodecanethiol (DDT, ≥98%), hexadecyltrimethyl ammonium bromide (CTAB, 98%), silver nitrate (AgNO3, ≥99.0%), 11-mercaptoundecanoic acid (MUA, 95%), and 4-nitrophenol (4-NP, ≥99.5%) were purchased from Sigma Aldrich (St. Louis, Missouri (Mo), USA). The chemicals were used as received without further purification.
6
Wistar Rat Study on Contrast-Induced Nephropathy
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A total of 28 clean-grade male Wistar (WT) rats at 6~8 weeks of age and a weight of 180~200 g were purchased from Beijing Vital River Laboratory Animal Technology Co. Ltd. (Animal Certificate of Conformity: SCXK (Beijing) 2006–0008). The animals were adaptively fed in the clean animal room at the Experimental Animal Center of the Academy of Military Medical Sciences for 1 week. All procedures for animal experiments were performed in accordance with the protocols and related provisions of the Academy of Military Medical Sciences Laboratory Animal Committee.
Probucol was purchased from Qilu Pharmaceutical Co. Ltd. (China); NAC was purchased from Hainan Zambon Pharmaceutical Co. Ltd. (China), Mikara; and N-nitro and -L-arginine methyl ester (L-NAME) were purchased from Sigma Company (USA). The low-osmolar, nonionic contrast media agent (Iopromide) was obtained from Schering AG (Germany).
Superoxide dismutase(SOD) and malondialdehyde (MDA) kit were purchased from Nanjing Jiancheng Company (China); 8-OHdG antibody was purchased from Abcam Company (England); A Mindary BS480 analyzer and a COBAS701 biochemical analyzer were used. The microscopic image acquisition and analysis system was from Olympus Co., Japan. The Hitachi H-600 transmission electron microscope was obtained from Hitachi.
Probucol was purchased from Qilu Pharmaceutical Co. Ltd. (China); NAC was purchased from Hainan Zambon Pharmaceutical Co. Ltd. (China), Mikara; and N-nitro and -L-arginine methyl ester (L-NAME) were purchased from Sigma Company (USA). The low-osmolar, nonionic contrast media agent (Iopromide) was obtained from Schering AG (Germany).
Superoxide dismutase(SOD) and malondialdehyde (MDA) kit were purchased from Nanjing Jiancheng Company (China); 8-OHdG antibody was purchased from Abcam Company (England); A Mindary BS480 analyzer and a COBAS701 biochemical analyzer were used. The microscopic image acquisition and analysis system was from Olympus Co., Japan. The Hitachi H-600 transmission electron microscope was obtained from Hitachi.
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