Anti-Interleukin-4 IL4 Antibody Picoband®

Figure. Western blot analysis of IL-4 using anti-IL-4 antibody (RP1009).

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: Recombinant Human IL-4 Protein 0.5ng,

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-IL-4 antigen affinity purified polyclonal antibody (Catalog # RP1009) 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 IL-4 at approximately 14KD. The expected band size for IL-4 is at 14KD.

Protective effect of M2 macrophages induced by hUCMSCs-EVs on OA chondrocytes in vitro. A IL-1β-induced OA chondrocytes were co-cultured with the supernatant of M2 macrophages (M2S) induced by hUCMSCs-EVs, or platelet-rich plasma (PRP) for 48 h, relative mRNA expression of the key genes TNF-α, MMP13, SOX9, and ACAN was measured by quantitative RT-PCR analysis; the experiment was performed triplicate; *p < 0.05, **p < 0.01, ***p < 0.001. B Western blot was performed to evaluate the expression of TNF-α, MMP13, and IL-4 proteins in PBS, M2S, or PRP-treated OA chondrocytes; GAPDH was employed as the loading control; *p < 0.05, **p < 0.01, ***p < 0.001. C The influence of M2S or PRP on the viability of chondrocytes was detected by the cell live/death experiment; green represents live cells while red represents dead cells; Scale bar: 1 mm
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Serum levels of IP-10, IFN-γ, IL-4, and TGF-β1 as well as the IFN-γ/IL-4 ratio in chronic hepatitis B patients with or without fibrosis. The levels of serum IP-10, IFN-γ, IL-4, and TGF-β1 in CHB patients with or without liver fibrosis were determined by ELISA, and the IFN-γ/IL-4 ratio was calculated. ♦♦♦ Differs from controls (the F0 group), P < 0.05; ★ ★ ★ differs from mild or minimal fibrosis (the F1–2 group), P < 0.05; ▴ ▴ ▴ differs from moderate fibrosis (the F3–4 group), P < 0.05. ( a ) IP-10; ( b ) IFN-γ; ( c ) IL-4; ( d ) TGF-β1; ( e ) the IFN-γ/IL-4 ratio.
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Statistical analysis of the correlation between the serum IP-10 level or the IFN-γ/IL-4 ratio with liver fibrosis among chronic hepatitis B patients. Spearman’s correlation analysis of the association between ( a ) IP-10; ( b ) IFN-γ; ( c ) the IFN-γ/IL-4 ratio and TGF-β1. Spearman’s correlation analysis of the association between serum ( d ) IFN-γ; ( e ) IL-4; ( f ) the IFN-γ/IL-4 ratio and IP-10.
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ROC curve analysis for evaluating the sensitivity and specificity of the IP-10 level, IFN-γ/IL-4 ratio, or their combination to predict significant fibrosis among CHB patients. ( a ) ROC curve analysis for serum IP-10 (with the cut-of f value of 300 pg/mL), the serum IFN-γ/IL-4 ratio (with the cut off value of 1.8), and the combination of IP-10 and the IFN-γ/IL-4 ratio; ( b ) Specificity and sensitivity for IP-10, the IFN-γ/IL-4 ratio, and their combination to predict significant liver fibrosis among patients with CHB.
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Intrahepatic mRNA levels of IP-10, IFN-γ, and IL-4 in chronic hepatitis B patients with or without fibrosis. Real-time qRT-PCR was conducted to quantify the mRNA levels of intrahepatic IP-10, IFN-γ, and IL-4 in the CHB patients without or with fibrosis as described in the Materials and Methods section. The relative mRNA levels of intrahepatic IP-10, IFN-γ, and IL-4 were calculated by comparative Ct analysis after normalization for the quantity of GAPDH in the same samples and were represented as 2 - △ △ Ct values for controls (the F0 group), which were set equal 1. ♦♦♦ Differs from controls (the F0 group), P < 0.05; ★ ★ ★ differs from mild or minimal fibrosis (the F1–2 group), P < 0.05; ▴ ▴ ▴ differs from moderate fibrosis (the F3–4 group), P < 0.05. (a) IP-10; (b) IFN-γ; (c) IL-4.
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Intrahepatic protein expression of IP-10, IFN-γ, IL-4, TGF-β1, and α-SMA as well as the IFN-γ/IL-4 ratio in chronic hepatitis B patients with or without fibrosis. The protein expression of intrahepatic (a, b, c, and d) IP-10, (e, f, g, and h) IFN-γ, and (i, g, k, and l) IL-4. In addition, the protein levels of intrahepatic IP-10, IFN-γ, IL-4, TGF-β1, and α-SMA were quantified based on the value of integrated optical density (IOD) and represented as histograms, from which the IFN-γ/IL-4 ratio was calculated. ♦♦♦ Differs from controls (the F0 group), P < 0.05; ★ ★ ★ differs from mild or minimal fibrosis (the F1–2 group), P < 0.05; ▴ ▴ ▴ differs from moderate fibrosis (the F3–4 group), P < 0.05.
Index in PubMed under a CC BY license. PMID: 28067328