Anti-Phospho-N kappa-p65 (T254) RELA Antibody

Western Blot analysis of various cells using Phospho-NFκB-p65 (T254) Polyclonal Antibody

Western blot analysis of lysates from 293 cells treated with TNF-alpha, using NF-kappaB p65 (Phospho-Thr254) Antibody. The lane on the left is blocked with the phospho peptide.

Immunohistochemistry analysis of paraffin-embedded human breast carcinoma, using NF-kappaB p65 (Phospho-Thr254) Antibody. The picture on the right is blocked with the phospho peptide.

Expression levels of C3 protein and mRNA in the mid colon of C3 KO mice. ( A ) The expressions of C3 protein and mRNA in the mid colon were measured by applying Western blot and RT-PCR analysis, using anti-C3 antibody and C3 specific primers. After determining the intensity of each band using an imaging densitometer, relative levels of the C3 protein were calculated, based on the intensity of β-actin. The mRNA level of the C3 gene was calculated based on the intensity of β-actin as an endogenous control. Tissue samples were collected from 3 to 5 mice per group, and each lysate was analyzed in duplicate for Western blot and RT-PCR analysis (final n = 6–10). ( B ) Tissue distribution of C3 protein was analyzed in the mid colon of WT and C3 KO mice. The C3 protein-specific antibody-stained sections of the mid colon from the WT and KO mice were observed at 400× magnification using light microscopy. The large image in the right column is a magnified image of the rectangle in the left column. H&E-stained sections (low rectangle in left corner) were observed at 400× magnification using a light microscope. ( C ) The expressions of C3aR and CR1 protein in the mid colon were measured with Western blot analysis using anti-C3aR and CR1 antibodies. After determining the intensity of each band using an imaging densitometer, relative levels of C3aR and CR1 proteins were calculated, based on the intensity of β-actin. Tissue samples were collected from 3 to 5 mice per group, and each lysate was analyzed in duplicate for Western blot (final n = 6–10). Data are reported as the mean ± SD. * indicates p < 0.05 compared to the WT mice.
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Expression levels of MAPK signaling pathway components. ( A ) Expression levels of ERK, p-ERK, JNK, p-JNK, p38 and p-p38 proteins were determined by Western blot analysis using the specific primary antibody and HRP-labeled anti-rabbit IgG antibody. ( B ) Band intensities were determined using an imaging densitometer, and protein expressions were calculated relative to the intensity of β-actin. Tissue samples were collected from 3 to 5 mice per group, and each lysate was analyzed in duplicate for Western blot (final n = 6–10). Data are reported as the mean ± SD. * indicates p < 0.05 compared to the WT mice.
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Expression levels of members in the iNOS‑mediated COX‑2 induction pathway. ( A ) Expression levels of COX-2 and iNOS proteins were determined by Western blot analysis using specific primary antibody and HRP-labeled anti-rabbit IgG antibody. ( B ) Band intensities were determined using an imaging densitometer, and protein expressions were calculated relative to the intensity of β-actin. Tissue samples were collected from 3 to 5 mice per group, and each lysate was analyzed in duplicate for Western blot (final n = 6–10). Data are reported as the mean ± SD. * indicates p < 0.05 compared to the WT mice.
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Expression levels of members in the ASC-inflammasome pathway. ( A ) Expression levels of NLRP3, cleaved-Cas1/Cas1 and ASC proteins were determined by Western blot analysis using the specific primary antibody and HRP-labeled anti-rabbit IgG antibody. ( B ) Band intensities were determined using an imaging densitometer, and protein expressions were calculated relative to the intensity of β-actin. Tissue samples were collected from 3 to 5 mice per group, and each lysate was analyzed in duplicate for Western blot (final n = 6–10). Data are reported as the mean ± SD. * indicates p < 0.05 compared to the WT mice.
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Expression levels of members in the NF-κB signaling pathway. ( A ) Expression levels of NF-κB-p65 and IκB-α proteins were determined by Western blot analysis using specific primary antibody and HRP-labeled anti-rabbit IgG antibody. ( B ) Band intensities were determined using an imaging densitometer, and protein expressions were calculated relative to the intensity of β-actin. Tissue samples were collected from 3 to 5 mice per group, and each lysate was analyzed in duplicate for Western blot (final n = 6–10). Data are reported as the mean ± SD. * indicates p < 0.05 compared to the WT mice.
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Levels of pro-inflammatory and anti-inflammatory cytokines. ( A ) The levels of TNF and IL-1α transcripts in the total mRNA of colon tissue were measured by qRT-PCR analyses using sense and anti-sense primers set for TNF and IL-1α. Concentration of the TNF protein was measured in the serum of WT and C3 KO mice using the ELISA Kit. This assay detects concentrations as low as 3.5 pg/mL for TNF. ( B ) The levels of anti-inflammatory cytokines including TGF- \(\beta\) 1 and IL-10 transcripts in the total mRNA of colon tissue were measured by qRT-PCR analyses using the sense and anti-sense primer set for TGF-β and IL-10. The mRNA level of each gene was calculated based on the intensity of actin as an endogenous control. Tissue samples were collected from 3 to 5 mice per group, and each lysate was analyzed in duplicate for qRT-PCR (final n = 6–10). ( C ) Expression levels of IL-6 protein were determined by Western blot analysis using specific primary antibody and HRP-labeled anti-rabbit IgG antibody. Band intensities were determined using an imaging densitometer, and protein expressions were calculated relative to the intensity of actin. Tissue samples were collected from 3 to 5 mice per group, and each lysate was analyzed in duplicate for ELISA (final n = 6–10). The concentrations of IL-6 proteins were measured in the serum of WT and C3 KO mice using the ELISA Kit. This assay detects concentrations as low as 2 pg/mL for IL-6. Data are reported as the mean ± SD. * indicates p < 0.05 compared to the WT mice.
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Levels of C5 and its mediated inflammatory regulators. ( A ) Expression levels of thrombin and C5 proteins were determined by Western blot analysis using specific primary antibody and HRP-labeled anti-rabbit IgG antibody. ( B ) Band intensities were determined using an imaging densitometer, and protein expressions were calculated relative to the intensity of β-actin. Tissue samples were collected from 3 to 5 mice per group, and each lysate was analyzed in duplicate for Western blot (final n = 6–10). Data are reported as the mean ± SD. * indicates p < 0.05 compared to the WT mice.
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Alterations in neutrophil infiltration and leaky epithelium. ( A ) Expression levels of E-cadherin were determined by Western blot analysis using specific primary antibody and HRP-labeled anti-rabbit IgG antibody. Band intensities were determined using an imaging densitometer, and protein expressions were calculated relative to the intensity of β-actin. Tissue samples were collected from 3 to 5 mice per group, and each lysate was analyzed in duplicate for Western blot (final n = 6–10). ( B ) MPO activity for neutrophil level. MPO activity was measured in lysate of mid colon tissues using the MPO assay kit. Tissue samples were collected from 3 to 5 mice per group, and each lysate was analyzed in duplicate for ELISA (final n = 6–10). ( C ) The levels of four tight junction channels including ZO-1, Occludin, Claudin-1 and Claudin-4 transcripts in the total mRNA of colon tissue were measured by qRT-PCR analyses using sense and anti-sense primers set for ZO-1, Occludin, Claudin-1 and Claudin-4. ( D ) The levels of four tight junction channels including CFTR, Ano-1, Slc26A3 and Slc26A6 transcripts in the total mRNA of colon tissue were measured by qRT-PCR analyses using sense and anti-sense primers set for CFTR, Ano-1, Slc26A3 and Slc26A6. The mRNA level of each gene was calculated based on the intensity of actin as an endogenous control. Tissue samples were collected from 3 to 5 mice per group, and each lysate was analyzed in duplicate for qRT-PCR (final n = 6–10). Data are reported as the mean ± SD. * indicates p < 0.05 compared to the WT mice.
Index in PubMed under a CC BY license. PMID: 35105928