Rabbit polyclonal anti-TGF-β1, anti-Col1, anti-Col3, anti-Smad 2, anti-Smad3, anti-Smad4, and MMP14 were purchased from Gene Tex Corporation (Irvine, California, USA). Rabbit polyclonal anti-Smad7 and anti-TIMP1 were purchased from Abcam Corporation (Cambridge science park, England). Rat polyclonal anti-GAPDH and HRP were purchased from Immunoway Corporation (Plano, TX, USA). RIPA cracking liquid, PMSF, BCA protein quantitative kit, and SDS-PAGE gel kit were purchased from Solarbio Science & Technology Co. Ltd. (Beijing, China). Huangqi Shengmai Yin (HSY) was purchased from Nan Chang Ji Sheng pharmaceutical factory (Nanjing, China).
Anti smad7
Manufactured by Abcam
Sourced in United States, United Kingdom
Anti-Smad7 is a laboratory research tool used to study the Smad7 protein. Smad7 is a key regulator in the TGF-beta signaling pathway. Anti-Smad7 can be used to detect and quantify Smad7 expression in various biological samples.
Automatically generated - may contain errors
Lab products found in correlation
Anti smad3, by Abcam (4 mentions)
Anti p smad2, by Abcam (3 mentions)
Anti smad2, by Abcam (3 mentions)
Anti p38, by Abcam (3 mentions)
Anti p p38, by Abcam (3 mentions)
Anti rage, by Abcam (3 mentions)
Hrp conjugated secondary antibody, by Beyotime (3 mentions)
Beyoecl plus, by Beyotime (3 mentions)
Anti gapdh, by Abcam (3 mentions)
Anti p smad3, by Abcam (3 mentions)
Anti e cadherin, by Abcam (2 mentions)
4 6 diamidino 2 phenylindole (dapi), by Thermo Fisher Scientific (2 mentions)
Mmp14, by GeneTex (1 mentions)
Anti timp1, by Abcam (1 mentions)
Anti tgf β1, by GeneTex (1 mentions)
Phenyl methyl sulfonyl fluoride (pmsf), by Solarbio (1 mentions)
Bca protein quantitative kit, by Solarbio (1 mentions)
Sds page gel kit, by Solarbio (1 mentions)
Eclipse 80i microscope, by Nikon (1 mentions)
Gapdh antibody, by Cell Signaling Technology (1 mentions)
10 protocols using anti smad7
1
Molecular Profiling of Fibrosis Markers
2
Immunohistochemical Analysis of Smad7 Expression
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
After deparaffinization and antigenic recovery by heat in citrate buffer (pH 6.0), the sections were permeabilized with Triton x-100 0.2% in phosphate buffered solution (PBS) for 5 minutes, followed by incubation with blocking solution of 5% BSA in PBS at room temperature for 30 minutes. After washing, the slides were incubated at 4°C overnight with rabbit polyclonal anti-Smad7 (Abcam, Cambridge, UK) diluted 1:100. The sections were incubated with FITC-labeled anti-rabbit goat IgG (Sigma-Aldrich, St. Louis, USA) diluted 1:50 for 1 h, and stained with 4,6-diamidino-2-phenylindole (DAPI, Thermo Scientific, Waltham, USA) for 7 min, and mounted in glycerol. Controls were treated by omitting the primary antibody. Sections were examined with a Nikon Eclipse 80i microscope (Tokyo, Japan). At least ten microscopic fields were taken from each rat under × 400 magnification using the 488 nm filter for Smad7 and 540 nm for DAPI. The ratio of Smad7 positive cells in relation to all cells present in the peritoneum (stained with DAPI) was calculated for every single picture, and the mean ratio was established for each rat.
3
Western Blot Analysis of RAGE and Smad Signaling
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Protease inhibitor cocktail was used to isolate total proteins, after which aliquots (30 μg/μl) were separated by SDS-PAGE and transferred to PDVF membranes. The membranes were then incubated with the primary antibodies (anti-RAGE, anti-Smad2, anti-p-Smad2, anti-Smad3, anti-p-Smad3, anti-p38, anti-p-p38, anti-Smad7 and anti-GAPDH, all purchased from Abcam, Cambridge, MA, USA) overnight at 4 °C. Glyceraldehyde-3-phosphate dehydrogenase (GAPDH) was used as a loading control. The following day, the membranes were incubated with the chemiluminescent reagent BeyoECL Plus and HRP-conjugated secondary antibodies (Beyotime, Shanghai, China). The signals were detected and photographed using a Tanon 6100 Chemiluminescent Imaging System (Tanon, Shanghai, China).
4
Subcellular Protein Analysis in A549 Cells
5
Immunohistochemical Analysis of Pituitary Adenoma
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Immunohistochemistry was performed on 5-μm sections from paraffin-embedded pituitary adenoma blocks. After conventional xylene dewaxing and gradient ethanol hydration, the sectioned samples were placed in 0.01 mol/L sodium citrate buffer (pH 6.0), heated for antigen retrieval and then cooled to room temperature. The sections were then incubated in 0.3% hydrogen peroxide solution at room temperature for 30 min to inactivate endogenous peroxidase. Sections were subsequently blocked with 5% goat serum/0.3% Triton X for 1 h and then incubated with various primary antibodies overnight at 4°C. Next, the primary antibody was targeted with an appropriate HRP-conjugated secondary antibody (1:500) for 1 h. The reaction was visualized using DAB staining reagent (Zhongshan Golden Bridge, Beijing, China) to produce a brown product, and sections were counterstained with hematoxylin. Finally, each section was mounted in buffered glycerol, observed and photographed under a microscope (ZEISS Axio Vert. A1, Zeiss, Germany). For the immunohistochemical analyses, the following antibodies were used: anti-TGF-β1 (Abcam), anti-E-cadherin (Abcam), anti-N-cadherin (Abcam), anti-Epcam (Abcam), anti-vimentin (Abcam), anti-Smad3 (Abcam), and anti-Smad7 (Abcam).
6
TGF-β1 and bAP15 Signaling Pathway Analysis
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
TGF-β1 (209-16554) was purchased from Fujifilm Wako Pure Chemical Industries (Osaka, Japan), and bAP15 (11324) was purchased from Cayman Chemical (Ann Arbor, MI, USA). The following antibodies were used: anti-β-actin (sc-47778), anti-UCHL5 (sc-271002), anti-Vimentin (sc-6260; Santa Cruz Biotechnology, Dallas, TX, USA), anti-E-cadherin (610181; BD Bioscience, San Jose, CA, USA), anti-smad1 (6944), anti-Phospho-Smad1 (5753), anti-Smad2 (5339), anti-Phospho-Smad2 (3108), anti-Phospho-Smad2 (18338), anti-Smad3 (9523), anti-Phospho-Smad3 (9520), anti-Smad4 (38454), anti-smad5 (12534), anti-Phospho-smad1/5 (9516), anti-Phospho-smad1/5/9 (13820), anti-ERK (9102), anti-Phospho-ERK (9101), anti-AKT (4691), anti-Phospho-AKT (4060; Cell Signaling Technology, Danvers, MA, USA), and anti-smad7 (216428; Abcam Inc., Toronto, ON, Canada).
7
Western Blot Analysis of Signaling Proteins
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Protease inhibitor cocktail was used to isolate total proteins, after which aliquots (30 μg/μl) were separated by SDS-PAGE and transferred to PDVF membranes. The membranes were then incubated with the primary antibodies (anti-RAGE, anti-Smad2, anti-p-Smad2, anti-Smad3, anti-p-Smad3, anti-p38, anti-p-p38, anti-Smad7 and anti-GAPDH, all purchased from Abcam, Cambridge, MA, USA) overnight at 4°C. Glyceraldehyde-3-phosphate dehydrogenase (GAPDH) was used as a loading control. The following day, the membranes were incubated with the chemiluminescent reagent BeyoECL Plus and HRP-conjugated secondary antibodies (Beyotime, Shanghai, China). The signals were detected and photographed using a Tanon 6100 Chemiluminescent Imaging System (Tanon, Shanghai, China).
8
HDAC1 Modulation in IL-17A-induced Fibrosis
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
1.1 Reagents MRC5 cells were donated to the school of life sciences, Tianjin NanKai University, Tianjin, China. IL-17A was purchased from Sino biological. TSA purchased from MedChemexpress. HDAC1 kit was purchased from Shanghai Jiemei Gene Medicine Technology. The MTT kit was purchased from Beijing Solebo Technology. HDAC1, Vimentin, Anti-a-SMA, anti-HDAC1, anti-p-Smad2, anti-p-smad3, anti-Smad2 and anti-Smad3 were acquired from Cell Signaling Technology of USA. Anti-collagen-I and anti-Smad7 were purchased from Abcam.
Olympus cell sensentry was purchased from Olympus in Japan.
Olympus cell sensentry was purchased from Olympus in Japan.
9
Western Blot Analysis of Signaling Proteins
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Protease inhibitor cocktail was used to isolate total proteins, after which aliquots (30 μg/μl) were separated by SDS-PAGE and transferred to PDVF membranes. The membranes were then incubated with the primary antibodies (anti-RAGE, anti-Smad2, anti-p-Smad2, anti-Smad3, anti-p-Smad3, anti-p38, anti-p-p38, anti-Smad7 and anti-GAPDH, all purchased from Abcam, Cambridge, MA, USA) overnight at 4°C. Glyceraldehyde-3-phosphate dehydrogenase (GAPDH) was used as a loading control. The following day, the membranes were incubated with the chemiluminescent reagent BeyoECL Plus and HRP-conjugated secondary antibodies (Beyotime, Shanghai, China). The signals were detected and photographed using a Tanon 6100 Chemiluminescent Imaging System (Tanon, Shanghai, China).
10
Intestinal Barrier Permeability in Mice
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
SLAC Laboratory Animal Co.Ltd,Shanghai, China Offers 8-week-old, weighting between 18g and 22g, Speci c pathogen-free (SPF) grade C57BL/6J mice.Under 20±2℃ temperature,50% humidity and light/dark cycles of 12 h, mice were fed with tap water and standard pellet diet. Sigma-Aldrich Co Provided FITC-dextran 4000 (FD-4) and DSS,while the molecular weight of the latter is 8000. And the Nanjing Jiancheng Biotechnology Institute (Nanjing, China) supplied kits for the detection of Evans blue (EB) and Myeloperoxidase (MPO) . MLCK ELISA kit was purchased from RB (USA). All the antibodies below, including anti-ZO-1, anti-E-Cadherin, anti-Occludin, anti-MLCK and anti-Smad7 were bought from Abcam (Cambridge, UK). Anti-TGF-β antibody was obtained from Gene Tex (USA). The following instruments were applied in this experiment,including light microscope (Olympus; Japan), Ultraviolet spectrophotometer (752 N; Shanghai, China), transmission electron microscope (TEM; Hitachi; Japan), enzyme-labeling instrument (ELx800;USA), and RT-PCR instrument (LightCycler480; Roche; Switzerland) .
About PubCompare
Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.
We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.
However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.
Ready to get started?
Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required
Revolutionizing how scientists
search and build protocols!