- PA5-20953 - Invitrogen Antibodies PIAS3 antibody

Submitted references TRIM27 promotes IL-6-induced proliferation and inflammation factor production by activating STAT3 signaling in HaCaT cells.

Miao X, Xiang Y, Mao W, Chen Y, Li Q, Fan B
American journal of physiology. Cell physiology 2020 Feb 1;318(2):C272-C281

Protein Inhibitor of Activated STAT3 Suppresses Oxidized LDL-induced Cell Responses during Atherosclerosis in Apolipoprotein E-deficient Mice.

Wang R, Zhang Y, Xu L, Lin Y, Yang X, Bai L, Chen Y, Zhao S, Fan J, Cheng X, Liu E
Scientific reports 2016 Nov 15;6:36790

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Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Western blot analysis of K562 cell lysate using a PIAS3 polyclonal antibody (Product # PA5-20953) at 1 µg/mL.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Western Blot analysis of PIAS3 in K562 cell lysate with PIAS3 Polyclonal Antibody (Product # PA5-20953) at 1 µg/mL.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Western blot was performed using Anti-PIAS3 Polyclonal Antibody (Product # PA5-20953) and 76 kDa band corresponding to PIAS3 was observed across the cell lines tested. Modified whole cell extracts (1% SDS) (30ug lysate) of T-47D (Lane 1), PANC-1 (Lane 2) and RAW 264.7 (Lane 3) were electrophoresed using Novex® NuPAGE® 4-12 % Bis-Tris gel (Product # NP0322BOX). Resolved proteins were then transferred onto a nitrocellulose membrane (Product # IB23001) by iBlot® 2 Dry Blotting System (Product # IB21001). The blot was probed with the primary antibody (1ug/ml) and detected by chemiluminescence with Goat anti-Rabbit IgG (H+L) Superclonal™ Recombinant Secondary Antibody, HRP (Product # A27036, 1:4000 dilution) using the iBright FL 1000 (Product # A32752). Chemiluminescent detection was performed using SuperSignal™ West Dura Extended Duration Substrate (Product # 34076)..

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunohistochemistry of PIAS3 in human breast carcinoma tissue with PIAS3 Polyclonal Antibody (Product # PA5-20953) at 5 µg/mL.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Figure 2 IL-6 or ox-LDL stimulation reduces PIAS3 expression in RAW264.7 cells. ( A ) IL-6 treatment leads to reductions in PIAS3 expression levels in a time- and dose-dependent manner, as determined via RT-qPCR. ( B ) Detection of PIAS3, STAT3-Y705 and STAT3 protein expression in IL-6-treated cells via Western blotting. Full-length blots were presented in Supplementary Figure S2 . ( C ) Detection of IL-6 and PIAS3 expression in ox-LDL-stimulated cells via RT-qPCR. ( D ) Stimulation with ox-LDL reduces PIAS3 protein expression. PIAS3, STAT3-Y705, STAT3 and iNOS protein expression levels in ox-LDL-treated cells were detected via Western blotting. Full-length blots were presented in Supplementary Figure S3 . ( E ) Relative levels of NO in the plasma of ApoE -/- mice fed a 20-week chow or western diet. Fold changes in NO levels relative to NO levels in mice fed a chow diet are shown (n = 8). Regarding the RT-qPCR results, relative transcript levels are expressed as fold changes compared to transcript levels in mock-treated cells. Regarding the Western blotting results, the relative protein levels in treated cells compared with those in mock-treated cells are shown below the images. *P < 0.05, **P < 0.01, ***P < 0.001.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Figure 1 Atherosclerotic plaque formation is inversely correlated with PIAS3 expression in the aorta. ( A ) Representative images of Oil Red O-stained aortas. ( B ) Representative images showing atherosclerotic plaque formation at the aortic root (bar, 200 mum). ( C ) The relative sizes of the plaque areas at the aortic roots are expressed as fold changes relative to the sizes of the plaque areas of ApoE -/- mice fed a chow diet (n = 8). Serial frozen sections of aortic roots were subjected to Oil Red O staining, and plaque areas were assessed. ( D ) IL-6 and PIAS3 gene expression levels in aortic tissue were determined by RT-qPCR (n = 8). ( E ) PIAS3, STAT3-Y705, STAT3 and IkappaBalpha protein expression levels in aortic tissue were determined by Western blotting. Full-length blots were presented in Supplementary Figure S1 . ( F ) Densitometry analysis of the Western blotting results in panel E. The average protein levels after normalization with beta-actin are expressed as fold changes relative to the protein levels in WT mice fed a western diet. ( G ) Representative images of PIAS3 protein detected in macrophages (Mphi) in atherosclerotic lesions. Serial frozen sections of aortic roots were subjected to immunohistochemical staining with Moma2 (for staining Mphi) and PIAS3 antibodies. Oil Red O staining was also performed to visualize the plaques (bar, 100 mum). *P < 0.05, **P < 0.01, ***P < 0.001.

PA5-20953

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