The largest database of trusted experimental protocols

5 protocols using c myc

1

Protein Expression Analysis in Wound Healing

Check if the same lab product or an alternative is used in the 5 most similar protocols
After incubation with PBS, OA-RD17 (1 nM), lipopolysaccharide (LPS, 1 μg/mL) (Solarbio, China), specific TLR4 inhibitor (1 μg/mL), miR-632 mimic (50 nM), or miR-632 inhibitor (100 nM) for 24 h, respectively, cell lysates (RIPA: PMSF: phosphatase inhibitor = 100:1:1; RIPA and PMSF, Meilun Biotechnology, Dalian, China; phosphatase inhibitor, Roche, Shanghai, China) were used to extract total protein in keratinocytes and macrophages. Moreover, cell lysates were also used to extract total protein in wound tissues from SD rats. The extracted proteins were quantified using the Bradford method (BCA protein analysis kit, Meilun, Dalian, China). The protein samples were then separated by 10% sodium dodecyl sulfate–polyacrylamide gel electrophoresis (SDS-PAGE), electro-imprinted on polyvinylidene fluoride membranes, and recorded and analyzed quantitatively using the Bole exposure software system. Primary antibodies, including GAPDH, Lamin B1, P38, P-P38, ERK, P-ERK, JNK, P-JNK, IκB, P-IκB, P65, P-P65 (Affinity, China), GSK3β, β-catenin, Cyclin D1, c-MYC, and Vimentin (ZEN BIO, China) were used following the provided instructions.
+ Open protocol
+ Expand
2

Western Blot Analysis of Epithelial-Mesenchymal Transition and Apoptosis Markers

Check if the same lab product or an alternative is used in the 5 most similar protocols
Cells were harvested and lysed with RIPA and the protein concentration was detected by BCA protein assay (Solarbio,PC0020). Twenty mg total proteins were separated by 10% SDS-PAGE and transferred to PVDF membrane, then incubated with the primary antibodies against E-cadherin (BA0475, 1:1500, BOSTER, China), N-cadherin (BA0673, 1:1000, BOSTER, China), Vimentin (PB9359, 1:1500, BOSTER, China), Bax (380,709, 1:2000, ZenBio, China), Bcl2 (381702,1:2000ZenBio, China), Cleaved-Caspase-3p17 (341,034, 1:1000ZenBio, China), Cleaved-Caspase-8-p18 (251,941, 1:2000ZenBio, China), β-catenin (R22820, 1:2000 ZenBio, China), c-Myc (R22809, 1:2000 ZenBio, China), Caspase-3 (R23315, 1:2000 ZenBio, China), MAL2 (BS-7175r, 1:1500, BIOSS BIOSS, China), GAPDH (GB11002, 1:2000, Servicebio, China) overnight. The membranes were subsequently incubated with the appropriate HRP-conjugated secondary antibodies (purchased from proteintech, SA00001-2, 1:10000) for 1.5 h, and signals were visualized using an ECL detection system.
+ Open protocol
+ Expand
3

Protein Expression Analysis in Stem Cells

Check if the same lab product or an alternative is used in the 5 most similar protocols
The cells were lysed in ice-clod RIPA cell buffer (P0013B, Beyotime Biotechnology). The total protein was separated by 10% SDS-PAGE and was transferred into PVDF membrane. The primary antibodies were incubated for overnight at 4 °C. They were USP13 (R26051, ZENBIO, 1:500), Raf1 (251817, ZENBIO, 1:1000), Phospho-MEK1/2 (Ser217/221) (310050, ZENBIO, 1:1000), MEK1/2 (380797, ZENBIO, 1:1000), Phospho-ERK1 (Thr202/Tyr204)/ERK2 (Thr185/Tyr187) (301245, ZENBIO, 1:500), Klf4 (381633, ZENBIO, 1:1000), c-Myc (382809, ZENBIO, 1:1000), HA (3724, Cell Signaling Technology, 1:1000), Flag (SG110-26, GNI, 1:1000), Ub (SC-8017, Santa Cruz, 1:1000), Oct4 (SC-5279, Santa Cruz, 1:1000), Nanog (14295-1-AP, Proteintech, 1:1000), Sox2 (66411-1-Ig, Proteintech, 1:1000), P53 (345567, ZENBIO, 1:1000), CDK4 (11026-1-AP, Proteintech, 1:1000), β-TUBULIN (200608, ZENBIO, 1:2000), and CDK1 (19532-1-AP, Proteintech, 1:1000). The bands were analyzed with High-sig ECL kit (180-501, Tanon).
+ Open protocol
+ Expand
4

Immunohistochemical Analysis of Tumor Markers

Check if the same lab product or an alternative is used in the 5 most similar protocols
IHC staining was performed as previously described [22 (link)]. Briefly, subcutaneous tumors were excised and fixed in formalin for 48 h. Serial 4-µm sections were cut, deparaffinized, blocked, and incubated at 4 °C overnight with the corresponding primary antibody: Ki67 (1:200, Cell Signaling Technology, 9449), c-Myc (1:200, Zen BioScience, 43250), ODC1 (1:200, Proteintech, 28728-1-AP). Then, an anti-rabbit/mouse IgG-HRP-linked secondary antibody (Genetech, GK500710) was added for 1 h at room temperature, followed by incubation with 3-diaminobenzidine tetrahydrochloride at room temperature for about 2 min. Hematoxylin was used to stain the nuclei. Finally, representative field photographs were captured using a BX63 microscope (Olympus) and analyzed using ImageJ software.
+ Open protocol
+ Expand
5

Protein Immunoblotting for Signal Pathways

Check if the same lab product or an alternative is used in the 5 most similar protocols
Protein purification, sodium dodecyl sulfate-polyacrylamide gel electrophoresis, electrical transfer, and immunoblotting were implemented as per the instructions. Primary antibodies for CXXC4 (1:1000; cat. no. ab105400; Abcam, Cambridge, MA, USA), p-GSK3β (Ser9) (1:1000; cat. no. 310010; ZenBio, Inc., Chengdu, China), β-catenin (1:1000; cat. no. 250110; ZenBio, Inc., Chengdu, China), cyclinD1 (1:1000; cat. no. 380999; ZenBio, Inc., Chengdu, China), and c-myc (1:1000; cat. no. 380784; ZenBio, Inc., Chengdu, China) were used to bind the target proteins. All immunoreactive protein complexes were incubated with horseradish peroxidase–conjugated secondary antibodies until they were detected.
+ Open protocol
+ Expand

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

Sign up now

Revolutionizing how scientists
search and build protocols!