Tissues were fixed in 4% paraformaldehyde for 24 h at room temperature and embedded in paraffin. Tissue sections (5 µm) were then dewaxed, rehydrated in graded alcohol, and incubated with 0.3% hydrogen peroxide to block endogenous peroxidase activity. Subsequently, the sections were treated with rabbit monoclonal anti-PDGFRB (cat no. 3169, 1:500; Cell Signaling Technology, Inc., Danvers, MA, USA) and anti-cluster of differentiation (CD) 34 (cat no. 3569, 1:500; Cell Signaling Technology, Inc.) antibodies at 4°C overnight, followed by incubation with a secondary anti-mouse immunoglobulin G antibody (cat no. sc-3749, 1:500; Santa Cruz Biotechnology, Inc., Dallas, TX, USA) for 15 min at room temperature, followed by diaminobenzidine staining. Each step was conducted according to the manufacturers' protocols. Microvessel density (MVD) was assessed using light microscopy by counting vessel density in 5 high-power fields.
Anti pdgfrb
Manufactured by Cell Signaling Technology
Sourced in United States
Anti-PDGFRB is a primary antibody product from Cell Signaling Technology. It recognizes the PDGFRB (Platelet-Derived Growth Factor Receptor Beta) protein.
Automatically generated - may contain errors
Lab products found in correlation
Sa β gal kit, by Beyotime (1 mentions)
Mitosox mitochondrial superoxide indicator, by Yeasen (1 mentions)
Anti nampt, by Cell Signaling Technology (1 mentions)
Anti pdgfra, by Cell Signaling Technology (1 mentions)
Anti upa, by Cell Signaling Technology (1 mentions)
Anti klf4, by Cell Signaling Technology (1 mentions)
Anti β actin, by Cell Signaling Technology (1 mentions)
Murine leukemia virus reverse transcriptase, by Promega (1 mentions)
Anti c myc, by Cell Signaling Technology (1 mentions)
Ge imagequant las4000 mini, by GE Healthcare (1 mentions)
Anti gapdh antibody, by Cell Signaling Technology (1 mentions)
Ripa lysis buffer, by Cell Signaling Technology (1 mentions)
Pvdf membrane, by Merck Group (1 mentions)
Luminol reagent, by Thermo Fisher Scientific (1 mentions)
Protease inhibitor cocktail, by Roche (1 mentions)
Bca assay kit, by Keygen Biotech (1 mentions)
3 protocols using anti pdgfrb
1
Immunohistochemical Analysis of PDGFRB and CD34 in Tissues
2
Overexpression and Knockdown of Key Genes
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Adv-vectors (KLF4, PDGFRA and NAMPT) were used to overexpress target genes, and sh-RNAs (KLF4, PDGFRA and NAMPT) were used to knock down target genes. All were purchased from Viogene Bio. (Jinan, Shandong, China). The KLF4 luciferase reporter plasmid (pGL4-KLF4) was purchased from Yeasen Bio. (Shanghai, China). A fluorescent dual luciferase reporting system and murine leukemia virus reverse transcriptase were obtained from Promega Co. (Mannheim, Germany). The SA-β-gal-kit was obtained from Beyotime (Haimen, China). The MitoSOX Mitochondrial Superoxide Indicator was obtained from Yeasen (Shanghai, China). The anti-p21 antibody was obtained from HUABIO (Hangzhou, China). The anti-PDGF-BB, anti-PDGFRA, anti-PDGFRB, anti-KLF4, anti-NAMPT, anti-PAI-2, anti-uPA and anti-β-actin antibodies and secondary antibodies were obtained from Cell Signaling Technology (Danvers, MA, USA). Other reagents were purchased from Sigma (St. Louis, MO, USA).
3
Western Blot Analysis of PDGFRB, C-myc, and GAPDH
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HASMCs were lysed in RIPA lysis buffer (Cell Signaling Technology, Boston, MA, USA) supplemented with protease inhibitor cocktail (Roche, Basel, Switzerland). The total protein concentration was measured using a BCA assay kit (Keygen, Nanjing, China). Equal amounts of protein were separated by SDS-polyacrylamide gel electrophoresis, transferred to polyvinylidene difluoride (PVDF) membranes (Millipore, Bedford, MA, USA), and blotted using anti-PDGFRB, anti-C-myc, and anti-GAPDH antibodies (Cell signaling Technology, Boston, MA, USA). The bands were visualized using Luminol reagent (Thermo Pierce, Waltham, MA, USA) and imaged using a GE ImageQuant Las 4000 mini (GE, Fairfield, CT, USA).
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