Mice underwent transcardiac perfusion on day 3 after SAH and brains were stored and fixed in 4% paraformaldehyde. Frozen brain samples were sectioned into 30 μm coronal sections using microtome. Brain slices were then incubated with a blocking buffer (0.4% Triton X-100 TBS-T with 5% donkey serum) for 2 h at room temperature and then incubated with primary antibodies (rabbit anti-NF-kB, #8242, Cell Signaling, Danvers, MA, USA {1:100}; Goat Anti-Iba-1, ab5076, Abcam, MA, USA {1:500}; Goat Anti-GFAP, A-31553, ThermoFisher, Waltham, MA, USA {1:200}; Rat anti-CD68, MCA1957, BIO-RAD, CA, USA {1:500}), followed by incubation at room temperature with secondary antibodies (Donkey anti-Rabbit Alexa Flour 488, A32790, ThermoFisher {1:1000}; Donkey anti-Goat Alexa Fluor 647, A-21447, ThermoFisher {1:1000}; Donkey anti-Rat Alexa Fluor 488, A-21208, ThermoFisher {1:1000}) in TBS-T for 45 min. After rinsing, the brain sections were stained with DAPI and mounted on the slide. The images were taken using Nikon Confocal microscopy. The fluorescent signaling was analyzed using Image J.
Rabbit anti nf kb
Manufactured by Cell Signaling Technology
Sourced in United States
Rabbit anti-NF-kB is a primary antibody that recognizes the NF-kB (nuclear factor kappa-light-chain-enhancer of activated B cells) protein. NF-kB is a transcription factor that plays a critical role in regulating the immune response, inflammation, and cell survival. This antibody can be used to detect and study the expression and localization of NF-kB in various experimental systems.
Automatically generated - may contain errors
Lab products found in correlation
Alexa flour 488 donkey anti rabbit, by Thermo Fisher Scientific (1 mentions)
Alexa fluor 488 donkey anti rat, by Thermo Fisher Scientific (1 mentions)
Alexa fluor 647 donkey anti goat, by Thermo Fisher Scientific (1 mentions)
Confocal microscopy, by Nikon (1 mentions)
Guinea pig anti vglut1, by Synaptic Systems (1 mentions)
Mouse anti stat3, by Cell Signaling Technology (1 mentions)
Transfer apparatus, by Bio-Rad (1 mentions)
Rabbit anti gapdh, by Synaptic Systems (1 mentions)
Bicinchoninic acid bca assay kit, by Thermo Fisher Scientific (1 mentions)
Mouse anti p65, by Cell Signaling Technology (1 mentions)
Rabbit anti snap25, by Synaptic Systems (1 mentions)
2 protocols using rabbit anti nf kb
1
Immunofluorescent Analysis of Neuroinflammation in SAH
2
Protein Extraction and Western Blot Analysis
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For total protein extraction, samples were homogenized using the following lysis buffer composed by 1% sodium dodecyl sulfate (SDS), 10mM HEPES at pH 7.4 and 2 mM EGTA and protein concentration was estimated using Bicinchoninic Acid Assay (BCA) kit (Thermo Fischer Scientific) as 20μg. Proteins were loaded with 2x Loading Buffer (100 mM Tris-HCl at pH 6.8; 4% SDS; 20% Glycerol; 200 mM 2-Mercaptoethanol, 2 mg Bromophenol-Blue) and fractionated by SDS-PAGE, then transferred to a nitrocellulose membrane using a transfer apparatus according to the manufacturer’s protocols (Bio-Rad). Membranes were stained with the following primary antibodies: mouse anti-STAT3 (Cell Signaling, 1:1000), rabbit anti-STAT3 P-Tyrosine 705 (Cell Signaling, 1:1000), mouse anti-STAT3 P-Serine 727 (Cell Signaling, 1:1000), rabbit anti-GAPDH (Synaptic System, 1:4000), mouse anti-PSD95 (UC Davis/NIH NeuroMab Facility, CA; 1:1000), guinea pig anti-VGLUT1 (Synaptic System, 1:1000), rabbit anti-Shank2/3 (Synaptic System, 1:1000), mouse anti-GAP43 (Millipore, 1:1000), rabbit anti-NFKB (Cell Signaling; 1:1000), rabbit anti-SNAP25 (Synaptic System, 1:1000), mouse anti-p65 (Cell Signaling; 1:1000). Immunodetection was performed with Clarity Western ECL Substrate (Bio Rab) and analyzed through Chemidoc apparatus via ImageLab software (Bio-Rab).
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