Transforming growth factor β (TGF-β), an important profibrotic growth factor and vascular endothelial growth factor (VEGF), a key angiogenic mediator of pulmonary fibrosis were determined by immunohistochemistry in lung tissues as previously described [33 (link)]. Lung sections were incubated overnight with anti-TGF-β (Santa Cruz Biotechnology; 1:100 sc-130348 in PBS, v/v, MA, USA) and anti-VEGF polyclonal antibody (Santa Cruz Biotechnology; 1:100 sc-7269 in PBS, v/v, MA, USA). At the end of the incubation with the primary antibodies, the sections were washed with PBS and incubated with a secondary antibody (Santa Cruz Biotechnology, Dallas, TX, USA) for 1 h at room temperature. The reaction was revealed by a chromogenic substrate (brown DAB), and counterstaining with nuclear fast-red. Images were collected using a microscope and AxioVision software. For graphical display of densitometric analyses, % positive staining (brown staining) was measured by computer-assisted color image analysis (Nikon Eclipse Ci-L microscope). The percentage area of immunoreactivity (determined by the number of positive pixels) was expressed as a percentage (%) of the total tissue area (red staining) at 20× magnification.
Anti vegf polyclonal antibody
Manufactured by Santa Cruz Biotechnology
Sourced in United States
The Anti-VEGF polyclonal antibody is a laboratory reagent produced by Santa Cruz Biotechnology. It is designed to specifically recognize and bind to the Vascular Endothelial Growth Factor (VEGF) protein.
Automatically generated - may contain errors
4 protocols using anti vegf polyclonal antibody
1
Quantification of Pulmonary Fibrosis Markers
2
Quantifying VEGF and CD34 Expression in Lung Tissues
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Immunohistochemistry (IHC) staining for vascular endothelial growth factor (VEGF) and CD34 was measured in the lung tissues as previously described [62 (link)]. Briefly, lung sections (7 μm) were processed and incubated overnight with anti-VEGF polyclonal antibody (Santa Cruz Biotechnology; 1:500 #sc7269 in PBS, v/v, MA, USA) and anti-Caspase-3 (Santa Cruz Biotechnology; 1:500 #sc7272 in PBS, v/v, MA, USA). To verify antibody-binding specificity, control slices were incubated with only primary antibody or secondary antibody, neither of which gave positive staining. Images were collected using a Zeiss microscope and Axio Vision software. For graphic display of densitometric analyses, the % of positive staining (brown staining) was measured by computer-assisted color image analysis (Leica QWin V3, UK). The percentage area of immunoreactivity (determined by the number of positive pixels) was expressed as % of total tissue area (red staining) within five random fields at ×40 magnification.
3
Tumor Growth Inhibition by Eltrombopag and Gemcitabine
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Six-week-old BALB/c mice were purchased by the Shanghai University of Traditional Chinese Medicine Institutional Animal Care and Use Committees (Shanghai, China). 4T1 cells (5×105) resuspended in PBS were injected into the axillary subcutaneous tissue (five mice per group). When palpable tumors (∼150 mm3) were observed, the mice received intraperitoneal (i.p.) injections of eltrombopag (75 and 150 mg/kg body weight) dissolved in PBS, gemcitabine (75 mg/kg body weight) dissolved in PBS or vehicle control every three days. The tumor volumes and body weight were measured every two days, and the tumor volumes were calculated as described previously31 (link). At the termination of the experiment, the mice were euthanatized, and tumors were harvested.
IHC was performed using formalin-fixed, paraffin-embedded (FFPE) tumors sectioned at a thickness of 5 μm with monoclonal anti-CD31 antibody (Abcam, Cambridge, UK), polyclonal anti-F4/80 antibody (Santa Cruz Biotechnology, Santa Cruz, CA, USA) and polyclonal anti-VEGF antibody (Santa Cruz Biotechnology). CD31, F4/80 and VEGF staining represented microvessel, macrophage and VEGF protein density, respectively. Staining and imaging were performed as described previously32 (link),33 (link). Tumor section images were analyzed with ImagePro Plus software.
IHC was performed using formalin-fixed, paraffin-embedded (FFPE) tumors sectioned at a thickness of 5 μm with monoclonal anti-CD31 antibody (Abcam, Cambridge, UK), polyclonal anti-F4/80 antibody (Santa Cruz Biotechnology, Santa Cruz, CA, USA) and polyclonal anti-VEGF antibody (Santa Cruz Biotechnology). CD31, F4/80 and VEGF staining represented microvessel, macrophage and VEGF protein density, respectively. Staining and imaging were performed as described previously32 (link),33 (link). Tumor section images were analyzed with ImagePro Plus software.
4
Quantifying VEGF Expression in Cells
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
VEGF expression in the section was carried out using polyclonal anti-VEGF antibody (1 : 200 dilution, rabbit, Santa Cruz Biotechnology). Positive control used for VEGF was placenta and negative control was devoid of the primary antibody. Slides were analyzed by light microscopy to determine reaction positivity. In this analysis, patients were divided into three groups: grade I: less than 1/3 of the stained cytoplasm, grade II: 1/3 to 2/3 of the stained cytoplasm, and grade III: more than 2/3 of the stained cytoplasm.
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!