As described previously (Kang et al. 2013 (link)), cells were trypsinized and lysed by Pro-Prep protein extraction kit (iNtRON Biotechnology, Seongnam, Korea). Equal amounts of protein extracts (20 &g) were separated by sodium dodecyl sulfate–polyacrylamide gel electrophoresis and transferred to a nitrocellulose membrane (Invitrogen). Blots were blocked with 5% nonfat dry milk at room temperature. The blots were incubated with antibodies specific for angiopoietin-1 (ab183701, Abcam), angiopoietin-2 (ab155106, Abcam), phosphorylated Tie2 (Tyr992, #4221, Cell Signaling Technology), Tie2 (sc-9026), thrombospondin-1 (sc-59886) and &-actin (Santa Cruz Biotechnology) at specific dilution, followed by incubation with peroxidase-labeled secondary antibodies. Immunoreactive proteins were visualized using an enhanced chemiluminescence detection kit (Santa Cruz Biotechnology). Blot images were captured using ImageQuant LAS 4000 biomolecular imager (GE Healthcare Life Sciences). Blot intensities were quantified using ImageJ 1.48v software (http://imagej.nih.gov/ij ) (Schneider et al. 2012 (link)).
Sc 9026
Manufactured by Santa Cruz Biotechnology
Sourced in United States
Sc-9026 is a laboratory equipment product offered by Santa Cruz Biotechnology. It is designed to perform a core function within the scientific research process. No further details can be provided while maintaining an unbiased and factual approach.
Automatically generated - may contain errors
Lab products found in correlation
Enhanced chemiluminescence detection kit, by Santa Cruz Biotechnology (1 mentions)
Ab155106, by Abcam (1 mentions)
Nitrocellulose membrane, by Thermo Fisher Scientific (1 mentions)
Ab183701, by Abcam (1 mentions)
Sc 59886, by Santa Cruz Biotechnology (1 mentions)
Imagequant las 4000 biomolecular imager, by GE Healthcare (1 mentions)
Pro prep protein extraction kit, by iNtRON Biotechnology (1 mentions)
Ab102015, by Abcam (1 mentions)
Ab32575, by Abcam (1 mentions)
Enhanced chemiluminescence substrate, by Beyotime (1 mentions)
Ab37168, by Abcam (1 mentions)
Horseradish peroxidase conjugated secondary antibody, by Aspen (1 mentions)
Nitrocellulose membrane, by Pall Corporation (1 mentions)
Ripa buffer, by Beyotime (1 mentions)
Bca kit, by Beyotime (1 mentions)
Ab6586, by Abcam (1 mentions)
Ab1893, by Abcam (1 mentions)
Ab76055, by Abcam (1 mentions)
Ab171123, by Abcam (1 mentions)
Ab45010, by Abcam (1 mentions)
3 protocols using sc 9026
1
Angiopoietin-Tie2 Signaling Analysis
2
Western Blot Analysis of Angiogenic Factors
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The samples were homogenized and total proteins extracted using a radioimmunoprecipitation assay (RIPA) buffer (Beyotime Institute of Biotechnology, Haimen, China). A BCA kit (Beyotime Institute of Biotechnology) was used to determine the protein concentration. The proteins were loaded on sodium dodecyl sulfate (SDS) polyacrylamide gels (10%) (Aspen), transferred onto nitrocel-lulose membranes (Pall Corp., New York, NY, USA), and blocked in non-fat dry milk (5%) at room temperature to set for 2 h. The membranes were incubated overnight at 4°C with primary antibodies against Ang-1 (1:1,000; ab102015; Abcam), p-Tie-2 (1:1,500; sc-9026; Santa Cruz Biotechnology, Inc.), α-SMA (1:2,000; ab32575), collagen type IV (1:1,500; ab6586) and GAPDH (1:5,000; ab37168) (all from Abcam). Subsequently, the membranes were incubated with horseradish peroxidase-conjugated secondary antibody (Aspen) for 1 h, and finally the membranes were detected using an enhanced chemiluminescence substrate (Beyotime Institute of Biotechnology).
3
Immunohistochemical Analysis of Angiogenic Markers
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Primary antibodies were rabbit anti-pIκB (2895S, Cell Signaling Technology), rabbit anti-NFκB (8242, Cell Signaling Technology), rabbit anti-pNFκB (3033, Cell Signaling Technology), rabbit anti-GAPDH (5174, Cell Signaling Technology), sheep anti-BrdU (ab1893, Abcam), mouse anti-CD31 (ab9498, Abcam), rabbit anti-CD68 antibody (ab125212, Abcam), rabbit anti-VCAM-1 (ab134047, Abcam), rabbit anti-VEGFR2 (ab 45010, Abcam), mouse anti-E-Cadherin (ab 76055, Abcam), mouse anti-ICAM-1 (ab 171123, Abcam), rabbit anti-Tie-2 (sc-9026, Santa Cruz Biotechnology) and Goat anti-mGalectin-3 (AF1197, R&D Systems). Secondary antibodies were donkey anti-mouse IgG (sc-2314, Santa Cruz Biotechnology), donkey anti-rabbit IgG-HRP (7074, Cell Signaling Technology), donkey anti-sheep IgG-555 (A21436; Invitrogen) and rabbit anti-sheep IgG (6150-04, Southern Biotech).
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!