Western blotting was performed according to the standard protocol. Briefly, 100 μg of total cell lysate was subjected to electrophoresis, after transfer the following antibodies were used to detect proteins: anti-TAp73 (A300-126A) (Bethyl Laboratories, TX, USA), anti-BAX (N-20; Santa Cruz Biotechnology, Germany) anti-PUMA (ABC158; Merck, MA, USA), anti-HO-1 (H-105; Santa Cruz Biotechnology), anti-NRF2 (H-300; Santa Cruz Biotechnology), anti-Bid (FL-195; Santa Cruz Biotechnology, TX, USA), anti-PARP (F-2; Santa Cruz Biotechnology), anti-β-actin (A2228; Sigma-Aldrich).
Anti nrf2 h 300
Manufactured by Santa Cruz Biotechnology
Sourced in United States
The Anti-Nrf2 (H-300) is a primary antibody that recognizes the Nuclear Factor Erythroid 2-Related Factor 2 (Nrf2) protein. Nrf2 is a transcription factor that plays a key role in the regulation of cellular antioxidant defense mechanisms.
Automatically generated - may contain errors
Lab products found in correlation
Anti α tubulin mouse mab dm1a, by Merck Group (2 mentions)
Anti bax n 20, by Santa Cruz Biotechnology (1 mentions)
Anti tap73 a300 126a, by Fortis Life Sciences (1 mentions)
Anti ho 1 h 105, by Santa Cruz Biotechnology (1 mentions)
Anti β actin, by Merck Group (1 mentions)
Ez chip kit, by Merck Group (1 mentions)
Etomoxir, by Bio-Techne (1 mentions)
Anti taz v386, by Cell Signaling Technology (1 mentions)
Anti mcl 1 s19, by Santa Cruz Biotechnology (1 mentions)
Chemiluminescence kit, by Santa Cruz Biotechnology (1 mentions)
Anti α tubulin t6074, by Merck Group (1 mentions)
Polyvinylidene fluoride (pvdf), by Merck Group (1 mentions)
Blocking buffer, by Merck Group (1 mentions)
Horseradish peroxidase hrp conjugated secondary antibody, by Merck Group (1 mentions)
Immobilon tm p, by Merck Group (1 mentions)
Anti il 1β, by Santa Cruz Biotechnology (1 mentions)
Anti vegf, by Abcam (1 mentions)
Anti inos, by Abcam (1 mentions)
Anti β actin, by Abcam (1 mentions)
7 protocols using anti nrf2 h 300
1
Western Blotting Protocol for Protein Detection
2
Nrf2 Transcription Factor Binding
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Patient lymphoblasts were seeded at 0.5 × 106/ml in tissue culture flasks, and incubated with 0.1% DMSO, 5 µM dyclonine or 5 µM dimethyl-fumarate. About ∼10 × 106 cells were used for each condition. After 24 h, cells were treated with 18.5% paraformaldehyde to crosslink proteins to DNA. ChIP was performed using an EZ-Chip kit (EMD Millipore). After shearing by sonication, each cell lysate was split and incubated with either Anti-Nrf2 (#H-300) or anti-mouse IgG negative control (Santa Cruz Biotechnology, Inc.) and protein-G-conjugated agarose beads to immunoprecipitate cross-linked protein/DNA. Crosslinks were reversed by incubating at 65°C overnight, and DNA was purified with provided spin columns and buffers. DNA concentration was determined using a Nanodrop 2000c (Thermo Scientific Corp.), and then analyzed by quantitative real-time PCR.
3
Antibody Panel for Protein Analysis
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The following antibodies were used: anti-ATF4/CREB-2 (c-20) (Santa Cruz Biotechnology, Cat. No. sc-200); anti-eIF4E1 (P-2) (Santa Cruz Biotechnology, Cat. No. sc-9976); anti-eIF4E3 (Proteintech, Cat. No. 17282-1-AP); anti-FAM129A/Niban (Signalway Antibody, Cat. No. 21401-2); anti-Keap1 (H-190) (Santa Cruz Biotechnology, Cat. No. sc-33569); anti-Nrf2 (H-300) (Santa Cruz Biotechnology, Cat. No. sc-13032); anti-RhoE/Rnd3 clone 4 (EMD Millipore Corporation, Cat. No 05-723); anti-SQSTM1/p62 (Clone 3) mouse mAb (BD Transduction Laboratories, Cat. No. 610832); and anti-α-tubulin mouse mAb (DM1A) (EMD Millipore Corporation, Cat. No. CP06). Carfilzomib (Cat. No. A-1098) was obtained from Active Biochem; chloroquine (Cat. No. S4157) and GSK2656157 (Cat. No. S7033) were purchased from Selleck Chemicals; (S)-4-carboxyphenylglycine (Cat. No. 0323), CGP57380 (Cat. No. 2731), 4EGI-1 (Cat. No. 4800) and MG-132 (Cat. No. 1748) were ordered from Tocris Bioscience; etomoxir (Cat. No. 11969) was obtained from Cayman Chemical; and trigonelline hydrochloride (Cat. No. T5509-1G) was from Sigma-Aldrich.
4
Nrf-2 Translocation in Antioxidant Response
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
We explored the role of transcription factor Nrf-2 [Nuclear factor (erythroid-derived 2)-like 2] which promotes mRNA expression and activity of antioxidant enzymes. Translocation of Nrf-2 to the nucleus was evaluated by fluorescence microscopy. Briefly, EPCs were washed in PBS, fixed with 4% (wt/vol) formaldehyde in PBS for 30 min at room temperature. After washing with PBS, the chamber slides were incubated with 0.2% TRITON X-100 in PBS for 10 min and then blocked with 1% BSA in PBS for 1 hour, followed by overnight incubation with anti-Nrf-2 (H-300) (Santa Cruz Biotechnology, Santa Cruz, CA, USA) at room temperature. The chamber slides were washed extensively before incubation for 1 hour with a secondary mouse anti-rabbit IgG-FITC antibody (Santa Cruz Biotechnology) in dark. The cells were washed with PBS and viewed with a fluorescence microscope. We used DAPI as nuclei-specific dye.
5
Immunoblotting Assay for Cell Signaling
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The following antibodies were used: anti-ZO-1/TJP1 (ZO1-1A12) (Fisher Scientific, Cat. No. 33-9100); anti-TAZ (V386) (Cell Signaling Technology, Cat. No. 4883); anti-TEF-1(TEAD1) (Clone 31/TEF-1) (BD Biosciences, Cat. No. 610922); anti-Nrf2 (H-300) (Santa Cruz Biotechnology, Cat. No. sc-13032); anti-MCL1 (S-19) (Santa Cruz Biotechnology, Cat. No: sc-819); and anti-α-tubulin mouse mAb (DM1A) (EMD Millipore Corporation, Cat. No. CP06). Carfilzomib (Cat. No. A-1098) was obtained from Active Biochem; emetine (Cat. No. 32-469-3250MG) was purchased from Fisher Scientific; and homoharringtonine (omacetaxine mepesuccinate) (Cat. No. SML1091) was from Sigma-Aldrich.
6
Western Blot Analysis of NRF2 Targets
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The western blot analysis from pancreatic cancer cells transfected with NRF2-siRNA or control-siRNA was carried out as described previously [31 (link)]. Briefly, proteins were separated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE), and transferred to polyvinylidene difluoride membranes (PVDF, Millipore, Bedford, MA, USA). Then, the membranes were blocked in 1X blocking buffer (Sigma, St. Louis, MO, USA) followed by incubation with the following primary antibodies: anti-NRF2 (H-300, Santa Cruz Biotechnology, Santa Cruz, CA, USA), anti-GCLC (ab53179), anti-ALDH1A1 (ab52492) and anti-ALDH3A1 (ab129022) (Abcam, Cambridge, UK) and anti-α-tubulin (T6074, Sigma, St. Louis, MO, USA). After washing three times with washing buffer, the membranes were incubated with horseradish peroxidase (HRP)-conjugated secondary antibodies (Sigma, St. Louis, MO, USA) and the signals were detected by the chemiluminescence kit (Santa Cruz Biotechnology).
7
Western Blot Analysis of Retinal Proteins
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Western blot analyses were performed using specific antibodies. Anti-Nrf2 (H-300) and Anti-IL-1β antibodies were purchased from Santa Cruz Biotechnology (Santa Cruz, CA, USA). Anti-iNOS, anti-VEGF, anti-glutamate cysteine ligase (GCL), and anti-β-actin antibodies were purchased from Abcam (Cambridge, MA). Other primary antibodies were purchased from Cell Signaling Technology (Beverly, MA, USA). Retinal samples were mechanically lysed in RIPA buffer (50 mM Tris-Cl, pH 7.5, 150 mM NaCl, 1% NP-40, 0.5% DOC, 0.1% SDS) with a cocktail of protease and phosphatase inhibitors (Sigma). Equal amounts of proteins were separated on 10% SDS polyacrylamide gels and transblotted onto polyvinylidene fluoride (PVDF) sheets (Immobilon TM-P, Millipore Corp., Bedford, MA, USA). Western signals were developed using HRP-conjugated secondary antibodies. Images were scanned from X-ray films and the band intensities were quantified with NIH Image J software.
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!