Anti-TNF alpha Antibody Picoband®

Western blot analysis of TNF alpha using anti-TNF alpha antibody (PB9010).
Electrophoresis was performed on a 5-20% SDS-PAGE gel at 70V (Stacking gel) / 90V (Resolving gel) for 2-3 hours. The sample well of each lane was loaded with 50ug of sample under reducing conditions.
Lane 1: PC-12 Whole Cell Lysate,
Lane 2: Rat Spleen Tissue Lysate,
Lane 3: Rat Brain Tissue Lysate,
Lane 4: Rat Kidney Tissue Lysate,
Lane 5: Rat Liver Tissue Lysate.
After Electrophoresis, proteins were transferred to a Nitrocellulose membrane at 150mA for 50-90 minutes. Blocked the membrane with 5% Non-fat Milk/ TBS for 1.5 hour at RT. The membrane was incubated with rabbit anti-TNF alpha antigen affinity purified polyclonal antibody (Catalog # PB9010) at 0.5 μg/mL overnight at 4°C, then washed with TBS-0.1%Tween 3 times with 5 minutes each and probed with a goat anti-rabbit IgG-HRP secondary antibody at a dilution of 1:10000 for 1.5 hour at RT. The signal is developed using an Enhanced Chemiluminescent detection (ECL) kit (Catalog # EK1002) with Tanon 5200 system. A specific band was detected for TNF alpha at approximately 26KD. The expected band size for TNF alpha is at 26KD.

Red nucleus IL-33 facilitates the early development of mononeuropathic pain by inducing TNF-α through activating ERK, p38 MAPK and JAK2/STAT3 signaling pathways. A Western blotting showed that red nucleus TNF-α was increased at 1 week post-SNI, intrarubral injection of anti-IL-33 antibody at 1 week post-SNI suppressed the overexpression of TNF-α ( n = 6 per group, F = 14.302, P < 0.001). B Western blotting displayed that intrarubral administration of PD98059, SB203580, or AG490 at 1 week post-SNI inhibited the production of TNF-α ( n = 6 per group, F = 13.157, P < 0.001). C Immunohistochemistry showed that red nucleus TNF-α was upregulated at 1 week post-SNI, intrarubral injection of anti-IL-33 antibody, PD98059, SB203580, or AG490 at 1 week post-SNI suppressed the production of TNF-α ( n = 4 per group, F = 33.029, P < 0.001). ** P < 0.01 and *** P < 0.001. Scale bars = 50 μm
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Red nucleus IL-33 evokes mechanical hypersensitivity by inducing TNF-α through activating ERK, p38 MAPK, and JAK2/STAT3 signaling pathways. A Western blotting indicated that intrarubral administration of IL-33 stimulated the secretion of TNF-α in naive rats ( n = 6 per group, F = 15.143, P < 0.001). B Western blotting showed that intrarubral pre-injection of PD98059, SB203580, or AG490, 30 min before IL-33 administration, restrained IL-33-induced overexpression of TNF-α in naive rats ( n = 6 per group, F = 9.812, P < 0.001). C Immunohistochemistry showed that intrarubral injection of IL-33 potentiated the secretion of TNF-α in naive rats, intrarubral pre-injection of PD98059, SB203580, or AG490, 30 min prior to IL-33 administration, inhibited IL-33-induced overexpression of TNF-α in naive rats ( n = 4 per group, F = 44.310, P < 0.001). *** P < 0.001. Scale bars = 50 μm
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DJ-1 interference increased the expression of TNF-α, IL-1β, and IL-6 after cerebral I/R injury. a and c Western blot detecting DJ-1 and the cytokines TNF-α, IL-1β, and IL-6 in rats. b , d Western blot detecting DJ-1 and the cytokines TNF-α, IL-1β, and IL-6 in astrocytes. e–g Quantification of TNF-α, IL-1β, and IL-6 in rats by ELISA. h‑j Quantification of TNF-α, IL-1β, and IL-6 in astrocytes by ELISA. The data are expressed as the mean ± SEM. * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001. n = 6 per group
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DJ-1 inhibited the expression of TNF-α, IL-1β, and IL-6 after cerebral I/R injury via SHP-1. a , c After virus and TPI-1 were used to overexpress DJ-1 and inhibit SHP-1, respectively, Western blotting was used to detect the cytokines IL-1β, IL-6, and TNF-α in rats. b , d After virus and TPI-1 were used to overexpress DJ-1 and inhibit SHP-1, respectively, Western blotting was used to detect the cytokines IL-1β, IL-6, and TNF-α in astrocytes. e , g After treatment with an SHP-1 inhibitor, Western blotting was used to detect the cytokines IL-1β, IL-6, and TNF-α in rats. f , h After treatment with an SHP-1 inhibitor, Western blotting was used to detect the cytokines IL-1β, IL-6, and TNF-α in astrocytes. i , k , m Quantification of TNF-α, IL-1β, and IL-6 in rats by ELISA. j , l , n Quantification of TNF-α, IL-1β, and IL-6 in astrocytes by ELISA. The data are expressed as the mean ± SEM. * p < 0.05 vs. the sham group; # p < 0.05 vs. the MCAO group; & p < 0.05 vs. the overexpression group; ★ p < 0.05 vs. the DMSO group; △ p < 0.05 vs. the DMSO group; ◆ p < 0.05 vs. the TPI-1 group. n = 6 per group. The data are expressed as the mean ± SEM. * p < 0.05 vs. the control group; # p < 0.05 vs. the OGD/R group; & p < 0.05 vs. the overexpression group; ★ p < 0.05 vs. the DMSO group; △ p < 0.05 vs. the DMSO group; ◆ p < 0.05 vs. the TPI-1 group. n = 6 per group. The groups in a and b and their corresponding groups in the statistical graphs are as follows: sham (−−−), MCAO or OGD/R (+−−), scramble (+−−), overexpression (++−), overexpression + DMSO (++−), and overexpression + TPI-1 (+++). The groups in e and f and the corresponding groups in the statistical graphs are as follows: DMSO (+−−), TPI-1 (+−+), overexpression + DMSO (++−), overexpression + TPI-1 (+++)
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DJ-1 regulated the disassociation of NLRX1 from TRAF6 after cerebral I/R injury via SHP-1. a , g After virus and TPI-1 were used to overexpress DJ-1 and inhibit SHP-1, respectively, Western blotting was used to detect NLRX1, TRAF6, and SHP-1 in rats. b , h After virus and TPI-1 were used to overexpress DJ-1 and inhibit SHP-1, respectively, Western blotting was used to detect NLRX1, TRAF6, and SHP-1 in astrocytes. c , i After treatment with an SHP-1 inhibitor, Western blotting was used to detect the cytokines IL-1β, IL-6, and TNF-α in rats. d , j After treatment with an SHP-1 inhibitor, Western blotting was used to detect the cytokines IL-1β, IL-6, and TNF-α in astrocytes. e Immunoprecipitation and immunoblot analyses of NLRX1-TRAF6 in rats. f Immunoprecipitation and immunoblot analyses of SHP-1-TRAF6 in rats. The data are expressed as the mean ± SEM. * p < 0.05 vs. the sham group; # p < 0.05 vs. the MCAO group; & p < 0.05 vs. the overexpression group; ★ p < 0.05 vs. the DMSO group; △ p < 0.05 vs. the DMSO group. n = 6 per group. The data are expressed as the mean ± SEM. * p < 0.05 vs. the control group; # p < 0.05 vs. the OGD/R group; & p < 0.05 vs. the overexpression group; ★ p < 0.05 vs. the DMSO group; △ p < 0.05 vs. the DMSO group. n = 6 per group
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Fecal microbiota transplantation (FMT) reduced systemic or local inflammatory response. (A) Protein levels of the pro-inflammatory cytokines and chemokines in the blood. (B) mRNA levels of the pro-inflammatory cytokines and chemokines in kidney tissue using real-time PCR. (C) Western blot analysis of the pro-inflammatory cytokines in kidney tissue. (D) Protein band intensities quantified using optical densitometry and normalized to β-actin levels within the respective experimental groups. n = 6. Representative images from six independent experiments. The p -values above the data denote statistical comparisons on the groups—the control (Ctrl), ischemia–reperfusion injury plus normal saline (IRI+NS), and IRI+FMT groups—which were calculated using ANOVA with Tukey’s post-hoc test. * p < 0.05; *** p < 0.001.
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Propionic acid reduced the systemic or local inflammatory response. (A) Protein levels of the pro-inflammatory cytokines and chemokines in the blood. (B) mRNA levels of the pro-inflammatory cytokines and chemokines in kidney tissue using real-time PCR. (C) Western blot analysis of the pro-inflammatory cytokines in kidney tissue. (D) Protein band intensities quantified using optical densitometry and normalized to β-actin levels within the respective experimental groups. n = 6. Representative images from six independent experiments. The p -values above the data denote statistical comparisons of the groups—the control (Ctrl), ischemia–reperfusion injury plus normal saline (IRI+NS), and IRI plus propionic acid (IRI+Prop) groups—which were calculated using ANOVA with Tukey’s post-hoc test. ** p < 0.01; *** p < 0.001.
Index in PubMed under a CC BY license. PMID: 41078364