Nonimmune goat serum was purchased from Fuzhou Maixin Biotech. Co., Ltd., (China). Anti-NF-kB p65 antibody was purchased from Abcam (Saudi Arabia). Anti-Rabbit IgG (H + L), F (ab') 2 Fragment was purchased from Cell Signaling Technology (USA). The EMSA kit was purchased from Thermo Fisher Scientific (USA). The natural protein extraction kit was purchased from MERCK company (German).
Anti nf kb p65 antibody
Manufactured by Abcam
Sourced in United States, United Kingdom
Anti-NF-kB p65 antibody is a lab equipment product that specifically recognizes the p65 subunit of the NF-kB transcription factor. It can be used to detect and quantify the expression levels of this protein in various biological samples.
Automatically generated - may contain errors
Lab products found in correlation
Anti occludin antibody, by Abcam (2 mentions)
Hrp conjugated goat anti rabbit secondary antibody, by Abcam (2 mentions)
Anti rage antibody, by Abcam (2 mentions)
Anti zo 1 antibody, by Thermo Fisher Scientific (2 mentions)
Pvdf membrane, by Thermo Fisher Scientific (2 mentions)
Image lab software, by Bio-Rad (2 mentions)
Ripa lysis buffer, by Beyotime (2 mentions)
Anti gapdh antibody, by Abcam (2 mentions)
Emsa kit, by Thermo Fisher Scientific (1 mentions)
Mini proten tetra system, by Bio-Rad (1 mentions)
Goat anti mouse igg, by Bioworld Technology (1 mentions)
Pvdf membrane, by Merck Group (1 mentions)
Chemidoc xrs system, by Bio-Rad (1 mentions)
Goat anti rabbit igg, by MultiSciences Biotech (1 mentions)
β actin, by MultiSciences Biotech (1 mentions)
Alexa fluor 488, by Thermo Fisher Scientific (1 mentions)
Iba1 polyclonal rabbit antibody, by Fujifilm (1 mentions)
Assay kit for gsh, by Merck Group (1 mentions)
Polyclonal rabbit antityrosine hydroxylase th antibody, by Novus Biologicals (1 mentions)
7 protocols using anti nf kb p65 antibody
1
NF-κB Activation Pathway Analysis
2
Western Blot Analysis of Tight Junction and Inflammatory Proteins
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Total proteins of cells were obtained by RIPA Lysis buffer (Beyotime Biotechnology, Jiangsu, China). The proteins were separated by 10% SDS-PAGE, transferred to polyvinylidene uoride (PVDF) membranes (Invitrogen, Carlsbad, CA, USA), and blocked with TBST containing 1.5% BSA at 26 ℃. After 2 h, the membranes were incubated with anti-ZO-1 antibody (1:1000, Invitrogen), anti-Occludin antibody (1:2000, Abcam), anti-RAGE antibody (1:500, Abcam), anti-NF-kB p65 antibody (1:1000, Abcam) and anti-GAPDH antibody (1:4000, Abcam), respectively, at 4°C overnight. After washed three times, the membranes were incubated with HRP-conjugated goat anti-rabbit secondary antibody (1:500, Abcam) at 37 ℃ for 2 h. The protein bands were visualized with ECL kit (Thermo Fisher Scienti c, Rockville, MD, USA), and images were taken with image lab software (Bio-Rad, Hercules, CA, USA).
3
Detecting NF-κB Expression by Western Blot
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Western blot analysis was used to detect NF-κB. A PVDF membrane was obtained from EMD Millipore (Billerica, MA, USA); cat. no. IPVH00010; batch no. K5JA5013L). Anti-NF-kB p65 antibody was obtained from Abcam (Cambridge, MA, USA; cat. no. ab7970). β-actin was purchased from Multi Sciences (Lianke) Biotech, Co., Ltd. (cat. no. ab008-100). Goat anti-rabbit IgG was bought from Multi Sciences (Lianke) Biotech, Co., Ltd. (cat. no. GAR0072). Goat anti-mouse IgG was obtained from Bioworld Technology, Inc. (St. Louis Park, MN, USA; cat. no. BS12478). An electrophoresis system (Mini-Proten Tetra system) was purchased from Bio-Rad Laboratories, Inc. (Hercules, CA, USA). A gel imager was obtained from Bio-Rad Laboratories, Inc. (ChemiDoc XRS+ system).
4
Western Blot Analysis of Tight Junction and Inflammatory Proteins
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Total proteins of cells were obtained by RIPA Lysis buffer (Beyotime Biotechnology, Jiangsu, China). The proteins were separated by 10% SDS-PAGE, transferred to polyvinylidene uoride (PVDF) membranes (Invitrogen, Carlsbad, CA, USA), and blocked with TBST containing 1.5% BSA at 26 ℃. After 2 h, the membranes were incubated with anti-ZO-1 antibody (1:1000, Invitrogen), anti-Occludin antibody (1:2000, Abcam), anti-RAGE antibody (1:500, Abcam), anti-NF-kB p65 antibody (1:1000, Abcam) and anti-GAPDH antibody (1:4000, Abcam), respectively, at 4°C overnight. After washed three times, the membranes were incubated with HRP-conjugated goat anti-rabbit secondary antibody (1:500, Abcam) at 37 ℃ for 2 h. The protein bands were visualized with ECL kit (Thermo Fisher Scienti c, Rockville, MD, USA), and images were taken with image lab software (Bio-Rad, Hercules, CA, USA).
5
Immunohistochemical analysis of neuroinflammation
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Polyclonal rabbit anti-cyclo-oxygenase-2 (COX-2), anti-inducible nitric oxide synthase (iNOS), anti-glial fibrillary acidic protein (GFAP), anti-cannabinoid receptor type 2 (CB2) and anti-NF-kB p65 antibodies were purchased from Abcam, Cambridge, MA, USA. Anti-ionized calcium binding adaptor molecule-1 (Iba-1) polyclonal rabbit antibody was purchased from Wako Chemicals, Richmond, VA, USA. Polyclonal rabbit anti-tyrosine hydroxylase (TH) antibody was obtained from Novus Biologicals, Littleton, CO, USA. Alexa Fluor® 488 and 594 conjugated secondary goat anti-rabbit antibodies were purchased from Life Technologies, Grand Island, NY, USA. CB2 receptor antagonist, AM630 ([6-Iodo-2-methyl-1-[2-(4-morpholinyl) ethyl]-1H-indol-3-yl] (4-methoxyphenyl) methanone) was purchased from Tocris Bioscience, Ellisville, MO, USA. The lipid peroxidation kit for estimation of malondialdehyde (MDA) was obtained from North West Life science (Vancouver, WA, USA). The compounds: ROT, BCP and the assay kit for GSH were procured from Sigma-Aldrich, St. Louis, MO, USA. All the reagents used in the study were of analytical grade.
6
Investigating NF-kB Translocation in BV-2 Cells
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Immuno uorescence staining was performed to determine the nuclear translocation of the NF-kB p65 in LPS-exposed BV-2 cells. The nuclear location of the NF-kB p65 is marked by anti-NF-kB p65 antibodies (1:1000) (Abcam, Cambridge, UK).
7
NF-kB p65 Nuclear Translocation Assay
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Immuno uorescence staining was performed to determine the nuclear translocation of the NF-kB p65 in LPS-exposed BV-2 cells. The nuclear location of the NF-kB p65 is marked by anti-NF-kB p65 antibodies (1:1000) (Abcam, Cambridge, UK)..
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!