Cells were washed three times with a phosphate buffer solution (PBS). The cells were then incubated with primary rabbit anti-4-HNE (1:1000, Abcam, Cambridge, MA) antibody overnight at 4°C. Following three-time washes with PBS, cells were incubated with secondary Alexa Fluor488-conjugated anti-rabbit IgG (1:200, Abcam, Cambridge, MA) for 30 minutes at room temperature. The DAPI was added to counterstain the nucleus for 5 minutes and 50% glycerinum was used to block the medium. Stained cells were photographed under a fluorescence microscope (Olympus, Tokyo, Japan).
Anti 4 hne
Manufactured by Abcam
Sourced in United States, United Kingdom
Anti-4-HNE is a laboratory product that can be used to detect and quantify the presence of 4-Hydroxynonenal (4-HNE) in biological samples. 4-HNE is a key marker of lipid peroxidation and oxidative stress. This product provides a tool for researchers to measure this important analyte.
Automatically generated - may contain errors
Lab products found in correlation
Anti cleaved caspase 3, by Cell Signaling Technology (6 mentions)
Anti 8 ohdg, by Abcam (4 mentions)
Anti nf κb p65, by Cell Signaling Technology (3 mentions)
Pierce ecl kit, by Thermo Fisher Scientific (3 mentions)
Anti β actin, by Santa Cruz Biotechnology (2 mentions)
Anti neun, by Merck Group (2 mentions)
Anti th antibody, by Enzo Life Sciences (2 mentions)
Anti egr1, by Cell Signaling Technology (2 mentions)
Ab46545, by Abcam (2 mentions)
Anti β actin, by Merck Group (2 mentions)
Anti c caspase3, by Cell Signaling Technology (2 mentions)
Anti gpx1, by GeneTex (2 mentions)
Anti nrf2, by Abcam (2 mentions)
Anti ho 1, by Abcam (2 mentions)
Anti orm1, by Abcam (2 mentions)
Normal goat serum (ngs), by Vector Laboratories (2 mentions)
4 6 diamidino 2 phenylindole (dapi), by Vector Laboratories (2 mentions)
Anti inos, by Abcam (2 mentions)
Anti gapdh, by Cell Signaling Technology (2 mentions)
4 6 diamidino 2 phenylindole (dapi), by Merck Group (2 mentions)
42 protocols using anti 4 hne
1
Immunocytochemistry for 4-HNE Detection
2
Green Tea Polyphenols Therapeutic Study
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
GTPs (91.21% catechins and 71.72% EGCG) were purchased from Corona Science & Technology Development Co. Ltd. (Fu Zhou, China). Rabbit polyclonal antibodies (anti-MnSOD, anti-HMGCS2, anti-Nampt, and anti-catalase) were purchased from Santa Cruz Biotechnology Inc. (Santa Cruz, USA). Anti-FOXO3a and anti-SIRT3 antibodies were from Cell Signaling Technology Inc. (Danvers, USA). Anti-SOD2/MnSOD (acetyl K122) and anti-4-HNE antibodies were from Abcam Inc. (Cambridge, UK). Total cholesterol (TC), triglycerides (TG), high-density lipoprotein-cholesterol (HDL-C), low-density lipoprotein-cholesterol (LDL-C), and blood glucose were detected using kits from BioSino Bio-Technology & Science Inc. (Beijing, China). ELISA kits for the quantitative measurement of serum cystatin C and insulin were from Biovendor Inc. (Heidelberg, Germany) and Mercodia AB (Uppsala, Sweden). Creatinine and N-acetyl-β-D-glucosaminidase (NAG) assay kits were purchased from the Nanjing Jiancheng Bioengineering Institute (Jiangsu, China). DMEM/high glucose was purchased from Gibco Inc. (New York, USA). Lipofectamine 3000 transfection reagent was from Invitrogen Inc. (California, USA).
3
Immunohistochemical and Fluorescent Analysis of Kidney Sections
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Kidney sections (4 μm) were first heated in 60°C, then deparaffinized in xylene, rehydrated in graded alcohol, and treated with 0.01 M sodium citrate buffer and 3% H2O2 to block endogenous peroxidase activity (immunofluorescence did not require this step). Then, 10% goat serum was added to block nonspecific antigen binding for 40 min in 37°C and in next step kidney sections were incubated with primary antibodies like anti-KIM-1 (1 : 200, Bioss), anti-CD68 (1 : 200, Proteintech), anti-TNF-α (1 : 200, Proteintech), anti-4-HNE (1 : 50, Abcam), and anti-Ly6G (1 : 100, Abcam) overnight at 4°C. Samples were then incubated in secondary antibodies (HRP conjugated for IHC and fluorescently labeled for IF) for 1 h at 37°C. The TdT-mediated dUTP nick-end labeling (TUNEL) staining was performed using commercial kits according to the manufacturer's instructions (Beyotime Biotechnology, Shanghai, China). DCF, DHE, and JC-1 staining was performed using commercial kits following the manufacturer's instructions (Beyotime Biotechnology, Shanghai, China). Slides were imaged using a microscope (Leica, Germany) or a confocal laser scanning microscope (Zeiss, Germany).
4
Molecular Mechanisms of Anti-Inflammatory Agents
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
3-Amino-1, 2, 4-triazole (#A8056 ), N-acetyl-l-cysteine (#A7250) and Chloroquine (#C6628), were purchased from Sigma-Aldrich. Recombinant mouse TNF-α (#MTA00B) and IL-6 (#M6000B) enzyme-linked immunosorbent assay (ELISA) kits (Quantikine) for cytokine analysis were bought from R&D Systems (Minneapolis, MN). Anti-PMP70 (#sab4200181, Sigma-Aldrich), anti-Catalase (#ab16731, Abcam), anti-SQSTM1/p62 (#H00008878-M01, Abnova), anti-LC3 (#L8918, Sigma-Aldrich), anti-Pex5 (#GTX109798, GeneTex), anti-Pex7 (#20614-1-AP, Proteintech), anti-DBP (#TA308904, OriGene), anti-ACOX1 (#10957-1-AP, Proteintech), anti-UBXD8 (#NB100-1296, Novs), anti-HA (#ab130275, Abcam), anti-Pex1 (#13669-1-AP, Proteintech), anti-Pex16 (#14186-1-AP, Proteintech), nti-Pex3 (#247042, Abcam), anti-Pex9 (PA5-22129, Invitrogen), anti-Tomm20 (#ab56783, Abcam), anti-NF-κB (#sc-8008, Santa Cruz), anti-IκBα (#9242S, CST), anti-p-IκBα (#2859S, CST), anti-4-HNE (#ab46545, Abcam) and anti-β-actin (#sc-47778, Santa Cruz).
5
Immunohistochemical Analysis of Alzheimer's Pathology
6
Quantifying Oxidative Stress Markers
7
Immunohistochemical Analysis of Oxidative Stress Markers
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Histological sections were incubated for 48 h, at 4 °C, with rabbit polyclonal antibodies anti-CML (1:30; Cat # ab27684), anti-AT1 receptor (1:50; Cat # ab18801), anti-4-HNE (1:50; Cat # ab46545) (Abcam, Cambridge, UK) and anti-RAGE (1:10; Cat # 600-401-P67) (Rockland Immunochemical Inc, Limerick, PA, USA) diluted in 1% bovine serum albumin (BSA) and 0.5% Triton. Samples were washed in phosphate-buffered saline (PBS) and 0.005% Tween 20 and incubated for 60 min at room temperature with Alexa Fluor® 488-conjugated goat anti-rabbit IgG antibody (Invitrogen, Fisher Scientific Baltics UAB, Cat # A11008, Vilnius, Lithuania) diluted at 1:200 in a PBS solution containing 0.003% Evans blue dye. After washing with PBS and Tween 20, the sections were incubated for 15 min at room temperature with 4,6-diamidino-2-phenylindole, dihydrochloride (DAPI) diluted at 1:200 in PBS.
See theSupplementary Materials for further information.
See the
8
Neuroprotective Cryosectioning and Staining
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Brain slabs were cryoprotected by soaking in 20 and 30% sucrose solution at 4°C and sliced into 30μm coronal sections. Firstly, sections were floated in 0.1% chrome alum-gelatin solution and mounted on glass slides. After drying at room temperature, sections were processed for Nissl staining. Then standard immunohistochemistry was carried out to examine DA neurons damage using anti-TH antibody (ENZO, USA) and lipid peroxidation using anti-4-HNE (Abcam, USA).
9
Comprehensive Neuronal Tissue Analysis
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
At the end of the experiment, all experimental animals were anesthetized with urethane (1.5 g/kg, i.p.) and transcardially perfused with 0.9% normal saline followed by 4% paraformaldehyde in a 0.1 M sodium phosphate buffer. Brain samples were removed, postfixed for 1 h, and stored in 30% sucrose for cryoprotection at 4 °C. Brain sections were washed three times in 0.01 M PBS after 1.2% hydrogen peroxide incubation to block endogenous peroxide activity. Primary neurons fixed with 4% PFA and the sectioned brains were incubated with phospho-AMPK (Abcam, 1:500), anti-MAP2 (Millipore, 1:500), anti-nNOS (ThermoFisher, 1:500), anti-4HNE (1:500), anti-nitrotyrosine (Abcam, 1:500), anti-phospho-TAU S396 (Abcam, 1:1000), anti-PSD95 (Invitrogen, 1:200), anti-IgG (Vector laboratories, 1:250), anti-SMI-71 (Covance, 1:500), anti-GFAP (Abcam, 1:1000), anti-AQP4 (Cell signaling, 1:500), anti-NeuN (Millipore, 1:500), anti-MMP9 (Abcam, 1:500), and anti-EGR1 (Cell signaling, 1:200) in a 4 °C incubator. Afterward, brain sections and cultured neurons were incubated with the fluorescence-conjugated secondary antibody for 2 h at room temperature.
10
Protein Expression Analysis in Lung Tissue
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
We isolated the cytoplasmic and nuclear fractions from the lung tissue and determined the protein content using a Bradford assay. Subsequently, lung protein lysates (30 μg per lane) were separated via 10%–12% sodium dodecyl sulfate-polyacrylamide gel electrophoresis. Immunoblots were then performed in accordance with the method described previously.9 (link)
The blots were subjected to probing with specific antibodies, including anti-ALDH2, anti-4-HNE (diluted at 1:1000, Abcam, Waltham, MA, USA), anti-PI3K, anti-AKT, anti-pAKT, anti-cleaved caspase 3, anti-NF-κB p65, anti-phospho-NF-κB p65, anti-IκB-α (diluted at 1:1000, Cell Signaling Technology, Danvers, MA, USA), anti-Bcl-2 (diluted at 1:1000, Bioss Inc., Woburn, Massachusetts, USA), anti-lamin B1 (diluted at 1:200, Santa Cruz Biotechnology, Dallas, TX, USA), or β-actin (diluted at 1:10,000, Sigma Chemical Company, St. Louis, MO, USA). The data were expressed by determining the relative ratio of the target protein's content to that of the reference protein. The relative ratio of the target protein's content in the control group was established as 1.
The blots were subjected to probing with specific antibodies, including anti-ALDH2, anti-4-HNE (diluted at 1:1000, Abcam, Waltham, MA, USA), anti-PI3K, anti-AKT, anti-pAKT, anti-cleaved caspase 3, anti-NF-κB p65, anti-phospho-NF-κB p65, anti-IκB-α (diluted at 1:1000, Cell Signaling Technology, Danvers, MA, USA), anti-Bcl-2 (diluted at 1:1000, Bioss Inc., Woburn, Massachusetts, USA), anti-lamin B1 (diluted at 1:200, Santa Cruz Biotechnology, Dallas, TX, USA), or β-actin (diluted at 1:10,000, Sigma Chemical Company, St. Louis, MO, USA). The data were expressed by determining the relative ratio of the target protein's content to that of the reference protein. The relative ratio of the target protein's content in the control group was established as 1.
About PubCompare
Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.
We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.
However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.
Ready to get started?
Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required
Revolutionizing how scientists
search and build protocols!