The corresponding ELISA kits were used to determine the cardiac contents of p-NF-κB p65 (Ser536) (Abcam, Cambridge, UK; Cat# ab176647), p-ERK1/2 (Thr202/Tyr204) (Invitrogen, CA, USA; Cat # EMS2ERKP), p-Akt (Ser473) (Elabscience, Wuhan, China; Cat # E-EL-R1135) and p-eNOS (Ser1177) (Cell Signaling Technology, MA, USA; Cat# 7980C).
P nf κb p65 ser536
Manufactured by Abcam
Sourced in United Kingdom
P-NF-κB p65 (Ser536) is a lab equipment product that detects the phosphorylation of the NF-kappa-B p65 subunit at serine 536. This modification is associated with the activation of the NF-kappa-B signaling pathway.
Automatically generated - may contain errors
Lab products found in correlation
P enos ser1177, by Cell Signaling Technology (1 mentions)
Donkey serum, by Merck Group (1 mentions)
Cd206, by Abcam (1 mentions)
P erk1 2 thr202 tyr204, by Cell Signaling Technology (1 mentions)
Lamin b, by Abcam (1 mentions)
Nf κb p65, by Abcam (1 mentions)
P38 mapk, by Abcam (1 mentions)
P ikkβ ser 180, by Cell Signaling Technology (1 mentions)
Cox 2, by Abcam (1 mentions)
Erk1 2, by Abcam (1 mentions)
β actin, by Abcam (1 mentions)
P p38 mapk thr180 tyr182, by Cell Signaling Technology (1 mentions)
3 protocols using p nf κb p65 ser536
1
ELISA-Based Cardiac Protein Profiling
2
Immunofluorescence Analysis of Activated Microglia
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The treated microglial cells were fixed with 4% paraformaldehyde at room temperature for 30 min and subsequently permeabilized with 0.2% Triton X-100. Following blocking with 10% donkey serum (Sigma-Aldrich; Merck KGaA) and 0.2% Triton X-100 at room temperature for 2 h, the samples were incubated at 4°C for 24 h with primary antibodies against IBA-1 (1:200; cat. no. ab5076; Abcam), iNOS (1:200; cat. no. ab15323; Abcam), CD206 (1:200; cat. no. ab8918; Abcam) and p-NF-κB p65 (Ser536) (1:200; cat. no. AF2006; Affinity Biosciences), followed by incubation with respective secondary antibodies at 37°C for 1 h, including 488-conjugated donkey anti-goat antibody (1:500; cat. no. E032231-02; Canlifesci, Inc.), 594-conjugated donkey anti-mouse antibody (1:500; cat. no. E032411-02; Canlifesci, Inc.), and 594-conjugated donkey anti-rabbit antibody (1:500; cat. no. E032421-01; Canlifesci, Inc.). Finally, the cells were stained with DAPI (cat. no. C1002; Beyotime Institute of Biotechnology) at room temperature for 10 min and subsequently examined with a scanning fluorescence microscope (DMi8; Leica Microsystems, Inc.).
3
Molecular Signaling Pathway Analysis
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ISO was purchased from Baxter Healthcare Corporation (Deerfield, IL, USA). Primary antibodies, including rabbit anti-rat JNK (cat#: ab76125), p38 MAPK (cat#: ab170099), ERK 1/2 (cat#: ab184699), IKKβ (cat#: ab124957), NF-κB p65 (cat#: ab16502), COX2 (cat#: ab15191), iNOS (cat#: ab3523), β-actin (cat#: ab179467), lamin B (cat#: ab16048), and p-NF-κB p65 (Ser536; cat#: ab76302) were acquired from Abcam (Cambridge, UK). Rabbit anti-rat phosphorylated (p)-JNK (Thr183/Tyr185; cat#: 4668), p-p38 MAPK (Thr180/Tyr182; cat#: 4511), p-ERK1/2 (Thr202/Tyr204; cat#: 4370), and p-IKKβ (Ser180; cat#: 3671) antibodies were procured from Cell Signaling Technology, Inc. (Beverly, MA, USA). Horseradish peroxidase (HRP)-conjugated anti-rabbit IgG (cat#: 0111-01) was provided by Chemicon (Temecula, CA, USA). NF-κB activation inhibitor (NAI; cat#: S1014-67.1) and p38 MAPK activation inhibitor (SB202190; cat#: 152121-30-7) were supplied by Biomol (Plymouth Meeting, PA, USA). ROS scavenger (NAC; cat#: A7250) and iNOS activity inhibitor (1400W; cat#: W4262) were purchased from Sigma–Aldrich (St. Louis, MO, USA). All the other reagents were obtained from Sigma unless specially stated. All suspensions were freshly prepared before use.
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