Total protein samples were collected from heart tissues or CFs. Lysate preparation: (RIPA: PMSF: protease inhibitor: phosphatase inhibitor = 100:1:2:2). The following reagents were used: RIPA (P0013C, Beyotime), PMSF (ST506, Beyotime), Protease and phosphatase inhibitor (P1045, Beyotime). The following primary antibodies were used: anti-α-SMA (Proteintech, 14,395-1-AP), anti-TGF-β1 (Proteintech,21,898-1-AP), anti-TGFBR1 (Abcam, ab235178), anti-collagen 1 (Col-1) (Proteintech,14,695-1-AP), anti-p-Smad3 (Cell Signaling Technology, #9520), anti-Smad3 (Cell Signaling Technology, #9523), and anti-GAPDH (Proteintech,60,004-1-lg). Goat anti-rabbit antibody (Ant Gene, ANT020) and Goat anti-mouse antibody (Ant Gene, ANT019) were applied as secondary antibodies.The following primary antibodies were used: anti-α-SMA (Proteintech, 14,395-1-AP), anti-TGF-β1 (Proteintech,21,898-1-AP), anti-TGFBR1 (Abcam, ab235178), anti-Col-1 (Proteintech,14,695-1-AP), anti-p-Smad3 (Cell Signaling Technology, #9520), anti-Smad3 (Cell Signaling Technology, #9523), and anti-GAPDH (Proteintech,60,004-1-lg). Goat anti-rabbit antibody (Ant Gene, ANT020) and Goat anti-mouse antibody (Ant Gene, ANT019) were applied as secondary antibodies.
Anti tgf β1
Manufactured by Proteintech
Sourced in United States, China
Anti-TGF-β1 is a laboratory reagent used for detecting and quantifying the presence of transforming growth factor beta 1 (TGF-β1) in biological samples. It functions as a specific antibody that binds to TGF-β1, which is a multifunctional cytokine involved in various cellular processes.
Automatically generated - may contain errors
Lab products found in correlation
Anti β actin, by Proteintech (5 mentions)
Anti gapdh, by Proteintech (5 mentions)
Anti smad3, by Cell Signaling Technology (3 mentions)
Anti vimentin, by Proteintech (3 mentions)
Anti akt, by Cell Signaling Technology (3 mentions)
Anti col 1, by Proteintech (3 mentions)
Anti n cadherin, by Cell Signaling Technology (2 mentions)
Anti smad2, by Cell Signaling Technology (2 mentions)
Anti p smad3, by Cell Signaling Technology (2 mentions)
Anti p akt ser473, by Cell Signaling Technology (2 mentions)
Anti e cadherin, by Proteintech (2 mentions)
Anti mmp9, by Proteintech (2 mentions)
Anti cd163, by Proteintech (2 mentions)
Anti inos, by Proteintech (2 mentions)
Ripa buffer, by Thermo Fisher Scientific (2 mentions)
Anti collagen 1, by Proteintech (2 mentions)
Fetal bovine serum (fbs), by Thermo Fisher Scientific (2 mentions)
Anti nicd, by Abcam (1 mentions)
Anti e cadherin, by Cell Signaling Technology (1 mentions)
Anti jagged1, by Santa Cruz Biotechnology (1 mentions)
17 protocols using anti tgf β1
1
Cardiac Fibroblast Protein Analysis
2
Antibody Characterization for Western Blot and Immunostaining
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Antibodies and reagents were obtained as follows: anti-Notch1 (western blot (WB), 1:800; immunohistochemical (IHC) or immunofluorescence (IF) staining, 1:400), anti-E-cadherin (WB and IF, 1:400), anti-N-cadherin (WB, 1:400), anti-α-SMA (WB, 1:800, IHC and IF, 1:400), and anti-Ki67 (IHC and IF, 1:400) antibodies from Cell Signalling Technology (CST, Beverly, MA, USA); anti-NICD (WB, 1:400, IHC and IF, 1:200), anti-Histone H3 (WB, 1:800), anti-c-Myc (WB, 1:800), and anti-vimentin (WB, 1:800; IHC, 1:400) antibodies from Abcam Company (Cambridge, MA, USA); anti-Jagged1 (WB, 1:800) antibody from Santa Cruz Biotechnology; anti-Smad2/Smad3 (phospho T8) (WB, 1:800) and anti-TGF-β1R (WB, 1:800) antibodies from MDL Biotechnology (Beijing, China); anti-GAPDH (WB, 1:8000), anti-Smad2/3 (WB and IF, 1:1000), anti-p-Smad2 (WB, 1:1000), anti-p-Smad3 (WB, 1:1000), anti-c-Myc (WB, 1:800); anti-collagen I (WB, 1:800; IF, 1:200), and anti-collagen III (WB, 1:800, IHC and IF, 1:200) antibodies from Biogot Technology (Shanghai, China); and anti-TGF-β1 (WB, 1:800) and anti-β-actin (WB, 1:8000) antibodies from Proteintech Biotechnology (Wuhan, China).
3
Western Blot Analysis of Lung Tissue
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Lung tissue was lysed by using RIPA buffer (Thermo Fisher Scientific). Equal amounts of proteins were separated by 10% sodium dodecyl sulphate‐polyacrylamide gel electrophoresis and transferred to polyvinylidene fluoride membranes (Bio‐Rad Laboratories). The membrane was blocked with 5% bovine serum albumin (Sigma‐Aldrich) for 90 min at room temperature and incubated with a primary antibody for 14 h at 4℃. The primary antibodies included anti–PPAR‐γ (Abcam), anti‐TGF‐β1 (ProteinTech), anti‐phosphor‐Smad3 (Cell Signaling Technology), anti‐Smad3 (Cell Signaling Technology), anti‐phosphor‐Stat6 (Cell Signaling Technology), anti‐Stat6 (Santa Cruz Biotechnology), anti‐phosphor‐Stat3 (Cell Signaling Technology), anti‐Stat3 (Santa Cruz Biotechnology), anti‐β‐tubulin (ProteinTech), anti‐NFκB P65 (Abcam), anti‐IκBα (Abcam), anti‐IL‐1β (Abcam), anti‐phosphor‐NFκB P65 (Cell Signaling Technology), anti‐phosphor‐IκBα (Cell Signaling Technology) and anti‐GAPDH (Abcam). And then, membranes were incubated with anti‐rabbit/mouse IgG (H+L) (Abcam) for 1 h at room temperature. Chemiluminescence measurement was performed by using Amersham Imager 680 (GE Healthcare Life Science).
4
Fibrosis Signaling Pathway Modulation
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BLM and the JNK inhibitor (SP600125) were obtained from Selleck China Inc. (Shanghai, China). TGF-β1 was purchased from PeproTech China Inc. (Suzhou, China). Azithromycin was obtained from Sigma-Aldrich Inc. (Shanghai, China). The primary antibodies we used are as follows: anti-vimentin (Proteintech, 60330-1-Ig), anti-alpha-smooth muscle actin (α-SMA) (Proteintech, 14395-1-AP), anti-Collagen 1 (Proteintech, 14695-1-AP), anti-LOX (Proteintech, 17958-1-AP), anti-LOXL2 (Abcam, 96233), anti-TGF-β1 (Proteintech, 21898-1-AP), anti-Smad2 (Cell Signaling Technology, 5339), anti-Smad3 (Cell Signaling Technology, 9523), anti-phospho (P)-smad2 (Cell Signaling Technology, 3108), anti-P-smad3 (Cell Signaling Technology, 9520), anti-JNK (Proteintech, 66210-1-Ig), anti-c-Jun (Proteintech, 66313-1-Ig), anti-P-JNK (Proteintech, 80024-1-RR), anti-P-cJun (Proteintech, 28891-1-AP), anti-α-tubulin (Proteintech, 66031-1-Ig), and anti-GAPDH (Proteintech, 60004-1-Ig). The dilution ratio of all antibodies was 1:1000.
5
Antibody Identification for Kidney Research
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Sources of antibodies and reagents were as follows:
Anti-AKT (Cell Signaling Technology, Danvers, MA, USA), anti-p-AKT (Ser473) (Cell Signaling Technology, Danvers, MA, USA), anti-p-ERK1/2 (Cell Signaling Technology, Danvers, MA, USA), anti-ERK1/2 (Cell Signaling Technology, Danvers, MA, USA), anti-p-mTOR (Cell Signaling Technology, Danvers, MA, USA), anti-mTOR (Cell Signaling Technology, Danvers, MA, USA), anti-E-cad (Cell Signaling Technology, Danvers, MA, USA), anti-Cadh16 (Proteintech, Chicago, Illinois, USA), anti-snail1(Cell Signaling Technology:3879s), anti-α-SMA (Boster, Wuhan, Hubei, China), anti-AQP1 (Proteintech, Chicago, Illinois, USA), anti-ATP (Proteintech, Chicago, Illinois, USA), anti-TGF-β1 (Proteintech, Chicago, Illinois, USA), anti-GAPDH (Proteintech, Chicago, Illinois, USA), anti-TRPC6 (Alomone, Jerusalem, Israel), anti-TRPC3 (Alomone:ACC-016), anti-mouse IgG (KeRui, Wuhan, Hubei, China), anti-rabbit IgG antibody (KeRui, Wuhan, Hubei, China), recombinant human TGF-β1 (Cell Signaling Technology, Danvers, MA, USA), HYP9 (MedChem, Shanghai, China), and type-2 collagenase (Worthington Biochemical Corporation, Lakewood, Colorado, USA).
DMEM/f12 and FBS were purchased from Invitrogen (Chicago, California, USA). The whole sagittal section of the kidney was scanned by Biossci Biotechnology Company (Wuhan, Hubei, China).
Anti-AKT (Cell Signaling Technology, Danvers, MA, USA), anti-p-AKT (Ser473) (Cell Signaling Technology, Danvers, MA, USA), anti-p-ERK1/2 (Cell Signaling Technology, Danvers, MA, USA), anti-ERK1/2 (Cell Signaling Technology, Danvers, MA, USA), anti-p-mTOR (Cell Signaling Technology, Danvers, MA, USA), anti-mTOR (Cell Signaling Technology, Danvers, MA, USA), anti-E-cad (Cell Signaling Technology, Danvers, MA, USA), anti-Cadh16 (Proteintech, Chicago, Illinois, USA), anti-snail1(Cell Signaling Technology:3879s), anti-α-SMA (Boster, Wuhan, Hubei, China), anti-AQP1 (Proteintech, Chicago, Illinois, USA), anti-ATP (Proteintech, Chicago, Illinois, USA), anti-TGF-β1 (Proteintech, Chicago, Illinois, USA), anti-GAPDH (Proteintech, Chicago, Illinois, USA), anti-TRPC6 (Alomone, Jerusalem, Israel), anti-TRPC3 (Alomone:ACC-016), anti-mouse IgG (KeRui, Wuhan, Hubei, China), anti-rabbit IgG antibody (KeRui, Wuhan, Hubei, China), recombinant human TGF-β1 (Cell Signaling Technology, Danvers, MA, USA), HYP9 (MedChem, Shanghai, China), and type-2 collagenase (Worthington Biochemical Corporation, Lakewood, Colorado, USA).
DMEM/f12 and FBS were purchased from Invitrogen (Chicago, California, USA). The whole sagittal section of the kidney was scanned by Biossci Biotechnology Company (Wuhan, Hubei, China).
6
Molecular Mechanisms in Epithelial-Mesenchymal Transition
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Hyp (purity over 98%, B20631) was bought from the Yuanye Biotechnology Co., Ltd. (Shanghai, China). Bleomycin sulfate (S1214) was obtained from Selleck (Shanghai, China). Primary antibodies used in our project were as follows: anti-p-AKT Ser473(Cell Signaling Technology, 4060, dilution 1:2000), anti-AKT(Cell Signaling Technology, 4691, dilution 1:1000), anti-GSK-3β (Cell Signaling Technology, 12456, dilution 1:1000), anti-p-GSK-3β Ser9 (Cell Signaling Technology, 9323, dilution 1:1000), anti-TGF-β1 (Proteintech, 21898-1-AP, dilution 1:1000), anti-SNAIL1 (Proteintech, 13099-1-AP, dilution 1:1000), anti-E-cadherin (Proteintech, 20874-1-AP, dilution 1:1000), anti-a-SMA (Proteintech, 14395-1-AP, dilution 1:1000), anti-vimentin (Cell Signaling Technology, 5741, dilution 1:1000), anti-fibronectin (Proteintech, 15613-1-AP, dilution 1:1000), anti-TWIST1 (Proteintech, 25465-1-AP, dilution 1:1000), anti-N-cadherin (Proteintech, 22018-1-AP, dilution 1:1000), anti-GAPDH (Servicebio Biotechnology Co., Ltd., GB12002, dilution 1:1000), and anti-collagen I (Abcam, ab34710, dilution 1:1000).
7
Comprehensive Protein Expression Analysis
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Western blotting was performed as described previously.13 (link) The specific primary anti-FSTL3 (Invitrogen, USA. Lot: SI2436032) and anti-β-actin antibodies (Invitrogen, USA. Lot: RI2265993); the E-cadherin, N-cadherin, MMP2, MMP9, and SMAD2/3 Antibody Sampler Kits; and the SMAD 1/5/9 Antibody Sampler Kit (Cell Signaling Technology, USA. Lot: 2, 12, 7, 9, 14, 2) were used. Anti-Vimentin, anti-TGF-β1 (Proteintech, China. Lot: 00017056, 00021336), and anti-BMP1 (Abcam, UK. Lot: GR232094-5) antibodies were also used. The primary (1:1000) and secondary antibodies purchased from Zhongshan Golden Bridge Biotechnology (Beijing, China. Lot: 205001014, 203700821, 20500927) (1:5000) were added for the binding reaction. Exposure was detected in a gel image processing system (ChemiDoc XRS+) to analyze the target/β-actin bands, and the relative amounts of protein were calculated.
8
Investigating Inflammatory Signaling Pathways
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Santa Cruz Biotechnology (Dallas, Texas, USA) provided the anti‐TIMD4 (Cat# SC‐390805), anti‐ADAM17 (Cat# SC‐390859), and TGF‐β1 (Cat# sc‐130348) antibodies. Affinity Biosciences (Cincinnati, Ohio, USA) provided the anti‐p‐NF‐κB (Cat# AF2006), anti‐IL‐6 (Cat# DF6087), anti‐IL‐10 (Cat# DF6894), and anti‐IL‐18 (Cat# DF6252) antibodies. ABclonal (Boston, MA, USA) provided anti‐NF‐κB (Cat# A16271), anti‐phosphorylated p38 MAPK (p‐p38 MAPK, Cat# AP0526), anti‐p38 MAPK (Cat# A14401), anti‐caspase‐1 (Cat# A0964), and anti‐IL‐1β (Cat# A16288) antibodies. Proteintech (Chicago, IL, USA) provided the anti‐TLR‐4 (Cat# 66350‐1‐Ig), anti‐TGF‐β1 (Cat# 21898‐1‐AP) antibodies, HRP‐conjugated GAPDH monoclonal antibody (Cat# HRP‐60004), and fluorescein‐conjugated AffiniPure Goat Anti‐Rabbit IgG (H+L) (Cat# SA00003‐2). MedChemExpress (Monmouth Junction, NJ, USA) provided TAPI‐1 (Cat# HY‐16657) and SB203580 (Cat# HY‐10256). Yiyuan Biotechnology (Guangzhou, China) provided ox‐LDL (Cat# YB‐002). Sigma–Aldrich (St. Louis, MO, USA) provided LPS (Cat# L2630).
9
Protein Expression Analysis in Cardiomyocytes
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Proteins were extracted from the cultured cardiomyocytes and heart tissue using RIPA buffer (Fudebio, Hangzhou, China) mixed with protease inhibitor (Fudebio, Hangzhou, China) (1:100) and quantified by BCA assay (Thermo Fisher, USA). The primary antibodies used were anti-β-MHC (1:1000, Abcam, USA), anti-ANP (1:500, Abcam, USA), anti-TGF-β1 (1:1000, Proteintech, USA), anti-PTEN-induced putative kinase-1 (PINK1) (1:1000, Santa Cruz, USA), anti-Parkin (1:1000, Abcam, USA), anti-caspase-3 (1:1000, Cell Signalling Technology, USA), anti-Bcl-2 (1:1000, Proteintech, USA), and anti-β-actin (1:3000, Proteintech, USA). The secondary antibodies used were goat anti-rabbit and anti-mouse IgG-HRP (1:3000, Fudebio, Hangzhou, China). Bands were detected with ECL substrate (Fudebio, Hangzhou, China) and visualized with a GeneGnome imaging system (Syngene Bioimaging). Densitometry was carried out with ImageJ (NIH).
10
Immunohistochemical Analysis of Inflammatory Markers
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For immunohistochemical staining, after antigen retrieval, the slides were blocked with a 5% BSA solution for 1 hour and incubated with mouse anti-CD68 (1:100, Invitrogen, USA), anti-iNOS (1:100, Abcam, UK), anti-CD163 (1:1000, Proteintech, USA), anti-TNFα (1:100, ZenBio, China), anti-IL1β (1:100, Proteintech, USA), anti-IL10 (1:100, Proteintech, USA), anti-TGFβ1 (1:100, Proteintech, USA), anti-MMP9 (1:100, Proteintech, USA), anti-OCN (1:100, Affinity, China), anti-HIF1α (1:100, Proteintech, USA) and anti-SDHB (1:100, Proteintech, USA) at 4 °C overnight. Goat anti-rabbit IgG (1:100, Servicebio, China) was used as a secondary antibody and the nuclei were stained using 2-(4-amidinophenyl)-6-indolecarbamidine dihydrochloride (DAPI, Servicebio, China). Images were captured using Aperio AT2 (Leica, German).
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