After incubation in normoxia or for 6, 12, or 24 h under hypoxia, cells were lysed and the protein concentration was determined (10). Proteins were separated on 10% polyacrylamide gels and transferred onto a nitrocellulose membrane. The monoclonal antibodies used for blotting were: anti-N-cadherin (mouse, 1:3000), anti-E-cadherin (mouse, 1:10,000) and anti-GAPDH (mouse, 1:100,000) (all from BD Transduction, San-Jose, CA, USA), anti-vimentin (mouse, 1:200; Santa Cruz Biotechnology, Santa Cruz, CA, USA), anti-Snail (rabbit, 1:500; Novus Biologicals, Little Town, CO, USA), anti-Twist (mouse, 1:50; Abcam, Cambridge, UK) and HRP-conjugated anti-rabbit and anti-mouse IgG (both 1:7000; Santa Cruz Biotechnology). Western blot signals were measured by densitometric scanning with an Azure C300 Intelligent Dark Box (Biosystems Inc., Dublin, CA, USA). Each Western blot was performed three times.
Anti snail
Manufactured by Novus Biologicals
Sourced in United States
Anti-Snail is a lab equipment product designed to aid in the detection and study of Snail proteins. It provides a specific and sensitive method for the identification and analysis of Snail, a transcription factor involved in various cellular processes.
Automatically generated - may contain errors
Lab products found in correlation
Anti e cadherin, by Cell Signaling Technology (2 mentions)
Anti n cadherin, by Abcam (2 mentions)
Anti twist, by Abcam (1 mentions)
Anti e cadherin, by BD (1 mentions)
Anti vimentin, by Santa Cruz Biotechnology (1 mentions)
Anti gapdh, by BD (1 mentions)
Hrp conjugated anti mouse and anti rabbit igg, by Santa Cruz Biotechnology (1 mentions)
Anti n cadherin, by BD (1 mentions)
Anti β actin, by Santa Cruz Biotechnology (1 mentions)
Omat x ray film, by Kodak (1 mentions)
Anti nephrin, by Santa Cruz Biotechnology (1 mentions)
Anti podocin, by Merck Group (1 mentions)
Anti β catenin, by R&D Systems (1 mentions)
Anti wnt3a, by Abcam (1 mentions)
Anti p rr, by Abcam (1 mentions)
Anti fibronectin, by Santa Cruz Biotechnology (1 mentions)
Anti vimentin, by Cell Signaling Technology (1 mentions)
Anti zeb1, by Cell Signaling Technology (1 mentions)
Anti twist, by Cell Signaling Technology (1 mentions)
Anti zeb2, by Abcam (1 mentions)
4 protocols using anti snail
1
Evaluation of Epithelial-Mesenchymal Transition Markers
2
Podocyte Protein Expression Analysis by Western Blot
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Western blot analysis was performed as we described previously. In brief, homogenates from cultured podocytes were prepared using sucrose buffer containing protease inhibitor. After boiled for 5 min at 95°C in a 5× loading buffer, 20 µg of total proteins were subjected to SDS-PAGE, transferred onto a PVDF membrane and blocked by solution with dry milk. Then, the membrane was probed with primary antibodies of anti-PRR (1∶1000, Abcam), anti-Wnt3a (1∶1000, Abcam), anti-β-catenin (1∶500, R&D system), anti-snail (1∶500, Novus), anti-nephrin (1∶100, Santa Cruz), anti-podocin (1∶1000, sigma) or anti-β-actin (1∶5000, Santa Cruz) overnight at 4°C followed by incubation with horseradish peroxidase-labeled IgG (1∶5000). The immunoreactive bands were detected by chemiluminescence methods and visualized on Kodak Omat X-ray films. Densitometric analysis of the images obtained from X-ray films was performed using the Image J software (NIH, Bethesda, MD, USA).
3
Evaluating EMT Markers in Prostate Cancer
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Since CTR is shown to induce EMT in prostate cancer cells, we examined the expression of epithelial markers E-cadherin and ZO-1 as well as mesenchymal markers vimentin and fibronectin in mouse prostate sections. The sources and concentrations of the antibodies were as follows: anti-Vimentin (1:200), anti-E-cadherin (1:200) from Cell Signaling; anti-fibronectin (1:200) from Santa Cruz; anti-zonula occludens 1 (1:200) from Abcam (Cambridge, UK). Mouse prostate sections were also tested for the presence of EMT markers such as snail and N-cadherin. The source and concentrations were: anti-snail (1:100) from Novus Biologicals and anti-N-cadherin (1:400), from Abcam.
4
Western Blot Analysis of EMT Markers
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The cellular proteins were extracted in RIPA buffer supplemented with protease inhibitor. The protein concentration was quantified using BCA protein assay and 20 µg of each protein sample was loaded and analyzed by 10% sodium dodecyl sulfate (SDS)-polyacrylamide gel electrophoresis (PAGE) system. Then, proteins were transferred to a PVDF membrane. The membrane was blocked in 5% BSA and probed with primary antibodies: Anti-ZEB2 (1:1,000; no. ab138222, Abcam), anti-E-cadherin (1:1,000; no. 14472, Cell signaling), anti-N-cadherin (1:1,000; no. ab18203, Abcam), anti-vimentin (1:1,000; no. ab92547, Abcam), anti-ZEB1 (1:1,000; no. 70512, Cell signaling), anti-Snail (1:1,000; no. IMG-6639A, Novus Biologicals), anti-Slug (1:1,000; no. 9585, Cell signaling), anti-Twist (1:1,000; no. 69366, Cell signaling) and anti-GAPDH (1:2,000; no. ab8245, Abcam). Then, the membrane was incubated with peroxidase-conjugated anti-mouse or anti-rabbit secondary antibody (1:2,000, nos. NEF822001EA; NEF812001EA, PerkinElmer). Immunoreactivity bands were detected by chemiluminescence and the intensity of the bands was quantified using Image Lab Software (Bio Rad, China).
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!