Anti-IL6 Antibody Picoband®

Western blot analysis of IL6 using anti-IL6 antibody (PB9005).
Electrophoresis was performed on a 5-20% SDS-PAGE gel at 70V (Stacking gel) / 90V (Resolving gel) for 2-3 hours.
Lane 1: recombinant rat IL6 protein 0.5 ng.
After electrophoresis, proteins were transferred to a nitrocellulose membrane at 150 mA 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-IL6 antigen affinity purified polyclonal antibody (Catalog # PB9005) 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 1:5000 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 IL6 at approximately 25 kDa. The expected band size for IL6 is at 25 kDa.

Inflammation response were reduced by NLRC4 siRNA after ICH. a – g , j Western blot assay to detect NLRC4, pNLRC4, Pro-IL-1β, IL-1β, Pro-caspase-1, caspase-1, Pro-IL-18, IL-18, TNF-α, and IL-6, ( k , l ) ELISA for IL–18 and IL–1β ( h , i ) Immunostaining for MPO-positive cells (×400 and ×200) at peri-hematoma area in sham, ICH, negative control siRNA, and NLRC4 siRNA groups at 72 h after ICH (six rats for each group). * P < 0.05, compared with ICH.
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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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Effects of acacetin on inflammation-related cytokines in cells with hypoxia/reoxygenation exposure. Western blots and mean relative level of IL-6 (A) , TLR-4 (B) , IL-10 (C) in neonatal rat cardiomyocytes without (control) or with hypoxia/reoxygenation (H/R) exposure in the absence (V, vehicle) or presence of 0.3, 1, or 3 μM acacetin. Western blots and mean relative level of IL-6 (D) , TLR-4 (E) , IL-10 (F) in H9C2 cardiomyoblasts with the treatment used in (A–C) . Data were expressed as mean ± SEM and analyzed by one-way ANOVA followed by the Bonferroni-test ( n = 5 individual experiments, ∗ P < 0.05, ∗∗ P < 0.01 vs. control; # P < 0.05 vs. hypoxia/reoxygenation alone).
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Effects of silencing Nrf2 on apoptosis- and inflammation-related proteins in cells with hypoxia/reoxygenation insult. Western blots and relative levels of Bcl-2 (A) , Bax (B) , and cleaved caspase-1 (C) in H9C2 cardiomyoblasts transfected with control siRNA or Nrf2 siRNA and subjected to hypoxia/reoxygenation insult in the absence (V, vehicle) or presence of 3 μM acacetin (Aca). Western blots and relative levels of IL-6 (D) , TRL-4 (E) , and IL-10 (F) in H9C2 cardiomyoblasts with the treatment used in (A–C) . Data were expressed as mean ± SEM and analyzed by one-way ANOVA followed by Bonferroni-test ( n = 5 individual experiments, ∗ P < 0.05, ∗∗ P < 0.01 vs. vehicle of control siRNA; ## P < 0.01 vs. control siRNA with acacetin).
Index in PubMed under a CC BY license. PMID: 29867499