Cells were washed 3 times in cold PBS and lysed in RIPA buffer. The proteins were separated on 10% SDS-PAGE gels and then blotted onto PVDF membranes. The membrane was blocked with 5% milk and incubated with anti-NOTCH1 antibody (Abcam, UK) overnight and then incubated with HRP-conjugated secondary antibody (Abcam, UK) for 2 h. The proteins were detected using an ECL reagent kit (Thermo Fisher Scientific, USA).
Anti notch1 antibody
Manufactured by Abcam
Sourced in United States
The Anti-Notch1 antibody is a laboratory research tool used to detect and study the Notch1 protein, which plays a crucial role in cell-cell communication and development. This antibody can be used in various techniques, such as Western blotting and immunohistochemistry, to identify and analyze the expression of Notch1 in different cell types and tissues.
Automatically generated - may contain errors
Lab products found in correlation
Pvdf membrane, by Abcam (2 mentions)
Ecl reagent kit, by Thermo Fisher Scientific (1 mentions)
Hrp conjugated secondary antibody, by Abcam (1 mentions)
Anti tgf β, by Abcam (1 mentions)
Ecl reagent, by Santa Cruz Biotechnology (1 mentions)
Anti slug, by Abcam (1 mentions)
Anti gapdh antibody, by Bioworld Technology (1 mentions)
Anti dclk1 antibody, by Abcam (1 mentions)
Pvdf membrane, by Thermo Fisher Scientific (1 mentions)
Anti e cadherin, by Abcam (1 mentions)
Anti vimentin, by Abcam (1 mentions)
Anti mmp2, by Abcam (1 mentions)
Anti mmp9, by Abcam (1 mentions)
Anti vimentin antibody, by Cell Signaling Technology (1 mentions)
Anti akt antibody, by Cell Signaling Technology (1 mentions)
Hrp labeled goat anti mouse igg, by Abcam (1 mentions)
Anti e cadherin antibody, by Cell Signaling Technology (1 mentions)
Anti p akt antibody, by Cell Signaling Technology (1 mentions)
Anti β actin antibody, by Abcam (1 mentions)
3 protocols using anti notch1 antibody
1
Western Blot Analysis of NOTCH1
2
Western Blot Analysis of EMT Markers
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Whole-cell lysates were collected using RIPA buffer. Proteins were separated using 10% SDS polyacrylamide gel, and the gels were transported to PVDF membranes (Thermo Fisher Scientific, CA, USA). The PVDF membranes were incubated with 5% skim milk in TBST at room temperature for 1 h. Later on, the PVDF membranes were probed with primary antibodies: anti-DCLK1 antibody (1:1000, Abcam, CA, USA), anti-Notch1 antibody (1:3000, Abcam, CA, USA), anti-E-cadherin (1:3000, Abcam, CA, USA), anti-Vimentin (1:3000, Abcam, CA, USA), anti-slug (1:1000, Abcam, CA, USA), anti-TGF-β (1:1000, Abcam, CA, USA), anti-MMP2 (1:1000, Abcam, CA, USA), anti-MMP9 (1:1000, Abcam, CA, USA) and anti-GAPDH antibody (1:3000, Bioworld, CA, USA) overnight at 4 °C. After that, the PVDF membrane was incubated for 1 h in secondary antibody anti-rabbit IgG second antibody (Abcam; ab150077) (1:5000) at room temperature for 1 h. Finally, the immunoreactivity was detected using ECL reagent (Santa Cruz Biotechnology).
3
Commercial Antibodies for Western Blot
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The commercial primary antibodies used in the present study are as follows: Anti-EDG4 antibody (cat. no. 135980; Abcam, Cambridge, MA, USA), anti-Notch1 antibody (cat. no. 3608; Cell Signaling Technology, Inc., Danvers, MA, USA), anti-Hes-1 antibody (cat. no. 11988; Cell Signaling Technology, Inc.), anti-Akt antibody (cat. no. 4691; Cell Signaling Technology, Inc.), anti-p-Akt antibody (cat. no. 13038; Cell Signaling Technology, Inc.), anti-E-Cadherin antibody (cat. no. A42; Cell Signaling Technology, Inc.), anti-vimentin antibody (cat. no. 5741; Cell Signaling Technology, Inc.), anti-F-actin antibody (cat. no. 130935; Abcam) and anti-β-actin antibody (cat. no. 608407; Thermo Fisher Scientific, Inc.). All the primary antibodies were diluted to 1:1,000. The secondary antibodies used in the present study were as follows: HRP-labeled goat anti-rabbit IgG (cat. no. 97051; Abcam) and HRP-labeled goat anti-mouse IgG (cat. no. 205719; Abcam). The secondary antibodies were diluted to 1:5,000.
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!