The cells were rinsed briefly with phosphate-buffered saline (PBS) and fixed for 20 min in 4% paraformaldehyde in 0.1 M phosphate buffer (pH 7.4) at room temperature. The cells were permeabilized for 10 min with 0.1% Triton X-100 in PBS, and blocked for 45–60 min with 4% bovine serum albumin in PBS at room temperature. Cells were incubated overnight at 4 °C with one of the following antibodies: anti-Oct4 (1:500; Abcam, Cambridge, MA, USA), anti-Sox2 (1:500; NB110-37235, Novus Biologicals, Littleton, CO, USA), anti-Nanog (1:500; Abcam, Cambridge, MA, USA), anti-c-Myc (1:250; bs-4963R, Bioss, Woburn, MA, USA), anti-Klf4 (1:250, bs-1064R, Bioss, Woburn, MA, USA). This was followed by incubation with the following secondary antibody: Alexa Fluor 488-labeled anti-rabbit IgG (1:500; Invitrogen, Carlsbad, CA, USA). Nuclei were counterstained using DAPI (1 mg/mL PBS; Invitrogen, Carlsbad, CA, USA).
Anti c myc
Manufactured by Bioss Antibodies
Sourced in China
Anti-c-Myc is a laboratory reagent used in protein detection and purification. It is an antibody that specifically recognizes the c-Myc protein tag, which is commonly used to facilitate the identification and isolation of recombinant proteins expressed in various cell systems.
Automatically generated - may contain errors
Lab products found in correlation
Anti β catenin, by Bioss Antibodies (2 mentions)
Alexa fluor 488 labeled anti rabbit igg, by Thermo Fisher Scientific (1 mentions)
Anti nanog, by Abcam (1 mentions)
Anti oct4, by Abcam (1 mentions)
Bs 4963r, by Bioss Antibodies (1 mentions)
Anti sox2, by Novus Biologicals (1 mentions)
Nb110 37235, by Novus Biologicals (1 mentions)
Anti klf4, by Bioss Antibodies (1 mentions)
Bs 1064r, by Bioss Antibodies (1 mentions)
4 6 diamidino 2 phenylindole (dapi), by Thermo Fisher Scientific (1 mentions)
Enhanced chemiluminescent reagent, by Biosharp (1 mentions)
Anti gapdh, by Wuhan Servicebio Technology (1 mentions)
Anti e cadherin, by Wuhan Servicebio Technology (1 mentions)
Anti vimentin, by Huabio (1 mentions)
Anti n cadherin, by Wuhan Servicebio Technology (1 mentions)
Ripa lysis buffer, by Beyotime (1 mentions)
Ripa lysis buffer, by Sangon (1 mentions)
Goat anti rabbit igg, by Bioss Antibodies (1 mentions)
Enhanced ecl chemiluminescence detection kit, by Vazyme (1 mentions)
Anti β actin, by Bioss Antibodies (1 mentions)
4 protocols using anti c myc
1
Immunofluorescence Staining of Stem Cell Markers
2
Protein Expression Analysis in Cancer Cells
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
AGS and HGC27 cells were lysed in RIPA lysis buffer (Beyotime, Shanghai, China). The membranes were blocked with 5% skimmed milk and incubated overnight at 4 ℃ with the following primary antibodies. The primary antibodies were as follows: anti-HMGB1 (1:1,000; Bioss, Beijing, China), anti-β-catenin (1:1,000; Bioss), anti-Wnt3a (1:1,000; Bioss), anti-c-Myc (1:1,000; Bioss), anti-E-cadherin (1:1,000; Servicebio), anti-N-cadherin (1:1,000; Servicebio), anti-Vimentin (1:1,000; Huabio, Hangzhou, China), anti-Snail (1:1,000; Huabio) and anti-GAPDH (1:4,000; GB11002, Servicebio) at 4 ℃ overnight. Then, the prepared membranes were incubated with secondary antibody (1:10,000, Bioss) for 2 h. Finally, the blots were visualized with an enhanced chemiluminescent reagent (Biosharp, Beijing, China).
3
Protein Expression Analysis by Western Blot
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Protein was extracted by RIPA Lysis buffer (Sangon, Shanghai, China), subjected to SDS-PAGE gel, and transferred to PVDF membranes. The membranes were incubated with anti-p-glycoprotein (anti-p-gp, 1:500, Bioss, Beijing, China), anti-myeloid cell leukemia-1 (anti-MCL-1, 1:1,000, Bioss), anti-FBN1 (1:1,000, Bioss), anti-β-catenin (1:2,000, Bioss), anti-c-Myc (1:1,000, Bioss) or anti-β-actin (1:2,000, Bioss). After incubating with Goat Anti-Rabbit IgG (1:20,000, Bioss), the signals were determined by Enhanced ECL Chemiluminescence Detection Kit (Vazyme, Nanjing, China).
4
Immunofluorescence analysis of p21, c-Myc, and NF-κB
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
FaDu cells were grown on slides, fixed in 4% paraformaldehyde in PBS for 15 min at RT and washed with PBS. Cells were permeabilized with 0.2% Triton X-100 in PBS for 10 min. Cells were blocked with 5% goat serum for 1 hour, and incubated with rabbit polyclonal anti-p21 (1:100; Bioss Antibodies), rabbit polyclonal anti-C-Myc (1:100; Bioss Antibodies), rabbit monoclonal anti-NFκB p65 (1:100, Abcam) overnight at 4°C. The next day cells were washed 3 times with PBS, and incubated with goat Alexa 555 conjugated anti-rabbit IgG (1:400, Abcam) for 1 hour at room temperature in the dark. Cells were mounted in 70% glycerol and images were taken by laser confocal microscopy (Fluo-View FV1000; Olympus, Japan).
Detection of the fluorescent intensity (FI) of FaDu cells stained with anti-p21 or anti-C-Myc antibodies were preformed under a laser scanning confocal microscope. Positive signals were analyzed as mean fluorescent intensity (MFI) using the FV10-ASW 4.0 software (Fluo-View FV1000; Olympus, Japan). In brief, 100 cells from each treatment group were analyzed in a blinded manner. All images were captured under the same camera settings.
Detection of the fluorescent intensity (FI) of FaDu cells stained with anti-p21 or anti-C-Myc antibodies were preformed under a laser scanning confocal microscope. Positive signals were analyzed as mean fluorescent intensity (MFI) using the FV10-ASW 4.0 software (Fluo-View FV1000; Olympus, Japan). In brief, 100 cells from each treatment group were analyzed in a blinded manner. All images were captured under the same camera settings.
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!