For assessing protein expression in kidney tissues or cultured cells, sections or cells adhered to coverslips were incubated with the primary FSP-1 (Abcam, ab197896), α-SMA (Sigma, A2547), HMGB1 (Cell Signaling, #3935), F4/80 (Abcam, ab16911), SP-A (Santa Cruz), or TGF-β1 antibody (Santa Cruz) at a 1:100 dilution overnight. Corresponding FITC-conjugated secondary antibodies (Sigma, F0257-anti-mouse or F0382-anti-rabbit) were applied for each individual primary antibody at a 1:100 dilution, and fluorescent photomicrographs were obtained at ×100 or 200 magnification.
Tgf β1 antibody
Manufactured by Santa Cruz Biotechnology
Sourced in United States
The TGF-β1 antibody is a laboratory reagent used for the detection and quantification of the transforming growth factor beta 1 (TGF-β1) protein in various biological samples. TGF-β1 is a multifunctional cytokine involved in the regulation of cell growth, differentiation, and other cellular processes.
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Lab products found in correlation
F4 80, by Abcam (1 mentions)
Hmgb1, by Cell Signaling Technology (1 mentions)
α sma, by Merck Group (1 mentions)
Proper horseradish peroxidase conjugated secondary antibodies, by Jackson ImmunoResearch (1 mentions)
β actin antibody, by Santa Cruz Biotechnology (1 mentions)
Ctgf antibody, by Santa Cruz Biotechnology (1 mentions)
Fibronectin antibody, by Santa Cruz Biotechnology (1 mentions)
P akt antibody, by Santa Cruz Biotechnology (1 mentions)
Akt antibody, by Santa Cruz Biotechnology (1 mentions)
P erk antibody, by Santa Cruz Biotechnology (1 mentions)
Ripa buffer, by Beyotime (1 mentions)
Anti cleaved caspase 3 asp175, by Cell Signaling Technology (1 mentions)
Mayer s hematoxylin, by Merck Group (1 mentions)
3 3 diaminobenzidine (dab), by Merck Group (1 mentions)
Vectastain elite abc kit, by Vector Laboratories (1 mentions)
Positive charged slides, by Thermo Fisher Scientific (1 mentions)
5 protocols using tgf β1 antibody
1
Immunofluorescent Profiling of Kidney Markers
2
Protein Expression Analysis in Fibroblasts
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Western blot was performed following previously published method [29 (link)] to analyze the changes of specified proteins. Briefly, primary skeletal muscle fibroblasts were lyzed with RIPA buffer (Beyotime Bio, Shanghai, China) supplemented with proteinease inhibitor and phosphatase cocktail (Sigma, St. Louis, MO) and total proteins (40 μg) were resolved on 8% SDS-PAGE gels and transferred onto PVDF membranes. The membranes were blocked in 5% nonfat milk in TBST (50 mM Tris, pH 7.5; 150 mM NaCl; 0.1% Tween 20) for 45 min, incubated with primary antibodies at 4 °C overnight, washed and incubated with proper horseradish peroxidase conjugated secondary antibodies (Jackson ImmunoResearch, West Grove, PA) at room temperature for 60 min before visualized with enhanced chemiluminescence (ECL) reagents (Pierce, Rockford, IL). The primary antibodies used were Col I antibody, a-SMA antibody, CTGF antibody, TGF-β1 antibody, Fibronectin antibody, p-AKT antibody, AKT antibody, p-ERK antibody, PKC antibody, p-PKC antibody and β-Actin Antibody (sc-47,778) were purchased from Santa Cruz Biotech (Shanghai, China).
3
Immunohistochemical Assessment of Apoptosis and TGF-β1
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Immunohistochemistry of cleaved caspase-3 and TGF-β1 was carried out in paraffin-embedded 4 μm thick kidney sections as described previously [12 (link)]. After quenching of endogenous peroxidase activity, sections were incubated with a polyclonal rabbit anti-cleaved caspase-3 (Asp175) (1 : 50; Cell Signaling Technology, Inc., Beverly, MA, USA) and a polyclonal anti-transforming growth factor-β1 (TGF-β1) antibody (Santa Cruz Biotechnology, Inc., Beverly, MA, USA, dilution 1 : 100) overnight at 4°C in a humid atmosphere. Thereafter, sections were processed using the Vectastain Elite ABC Kit (Vector Labs, Burlingame, CA, USA), following the manufacturer's protocol. Immunostaining was performed with 3,3′-diaminobenzidine (DAB) (Sigma-Aldrich), and sections were counterstained with Mayer's hematoxylin (Sigma-Aldrich). Negative controls included incubation with a nonspecific Ig of the same isotype as the primary antibody. The surface labelled by the antibodies was evaluated using quantitative image analysis as previously described [13 (link)].
4
Quantifying TGF-β1 in Lung Tumors
5
Immunohistochemical Analysis of TGF-β1 in Colon Sections
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For the immunohistochemical investigations, colon sections (4 µm thick) were mounted onto positive-charged slides (Thermo Fisher Scientific, Pittsburgh, PA, USA) and immunostaining was conducted according to the methods described by Ahmed and Ahmed [65 (link)]. Briefly, the sections were incubated in 3% H2O2 solution for 15 min following deparaffinization, rehydration, antigen retrieval and sealing. Next, they were blocked and incubated with TGF-β1 antibody (Santa Cruz Biotechnology, Santa Cruz, CA, USA) (1:200 dilution) at 4 °C overnight. After washing with phosphate-buffered saline, the sections with the peroxidase-labeled secondary antibody (1:200 dilution) were incubated for 30 min. The bound antibody complex was visualized by the reaction of 3,3-diaminobenzidine (DAB) substrate and counterstaining with hematoxylin. This method was applied according to the instructions of ABclonal Inc. Company, Wuhan, China. The immunohistochemically stained sections were examined by a light microscope at high power (×400). The positive reaction appeared brown in color. The integrated intensities of the TGF-β1 response were measured using the ImageJ program.
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