Western blot analysis was performed as previously described by us [23 (link)]. The following antibodies were used in the analysis: anti-EVI5 (Millipore, Billerica, MA, USA); anti-Emi1 and anti-TGF-β receptor II (Santa Cruz, CA, USA); anti-CyclinA2 (Proteintech, IL, USA); anti-pAkt (Ser473), anti-Akt, anti-Erk1/2, anti-pErk (Thr202/Tyr204), anti-CyclinD1, anti-MMP2, anti-p-Smad3, anti-Snail and anti-β-actin (Cell Signaling Technology, Danvers, MA, USA); anti-TGF-β receptor I (Abcam, London, UK); anti-N-cadherin and anti-Vimentin (BD Biosciences, USA); Anti-mouse and anti-rabbit secondary antibodies (Cell Signaling Technology, Danvers, MA, USA).
Anti cyclina2
Manufactured by Proteintech
Sourced in United States
Anti-CyclinA2 is a primary antibody that binds to and detects the CyclinA2 protein. CyclinA2 is a key regulator of cell cycle progression and plays an important role in the G1/S and G2/M transitions. The Anti-CyclinA2 antibody can be used to study the expression and localization of CyclinA2 in various cell and tissue samples.
Automatically generated - may contain errors
Lab products found in correlation
Anti cyclin b1, by Proteintech (3 mentions)
Anti cyclin d1, by Proteintech (2 mentions)
Anti n cadherin, by BD (1 mentions)
Anti cyclin d1, by Cell Signaling Technology (1 mentions)
Anti mmp2, by Cell Signaling Technology (1 mentions)
Anti vimentin, by BD (1 mentions)
Anti erk1 2, by Cell Signaling Technology (1 mentions)
Anti β actin, by Cell Signaling Technology (1 mentions)
Anti mouse and anti rabbit secondary antibodies, by Cell Signaling Technology (1 mentions)
Anti emi1, by Santa Cruz Biotechnology (1 mentions)
Anti tgf β receptor 2, by Santa Cruz Biotechnology (1 mentions)
Anti tgf β receptor 1, by Abcam (1 mentions)
Anti snail, by Cell Signaling Technology (1 mentions)
Anti p erk thr202 tyr204, by Cell Signaling Technology (1 mentions)
Anti p smad3, by Cell Signaling Technology (1 mentions)
Polyvinylidene fluoride (pvdf), by Cytiva (1 mentions)
Anti p21, by Proteintech (1 mentions)
β actin, by Proteintech (1 mentions)
Anti erk, by Proteintech (1 mentions)
Anti phospho erk1 2, by Proteintech (1 mentions)
4 protocols using anti cyclina2
1
Western Blot Analysis of EMT Signaling
2
Western Blot Analysis of GC Cell Proteins
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
In a radio-immunoprecipitation assay (RIPA) lysis buffer, the proteins were extracted from GC cells. Ten percents of polyacrylamide sodium dodecyl sulfate (SDS) gels (SDS-PAGE) were used to separate equivalent amounts of proteins. The proteins were then blotted onto membranes of polyvinylidene fluoride (Amersham, UK). After membranes were blocked at RT (37°C) in bovine serum albumin (5%) for 1 h, primary antibodies were used to culture the membranes overnight. After being cultured, they were placed in secondary antibodies conjugated with human resource planning (Perbio Science, Belgium), and incubated at RT (37°C) for 1 h. The system of enhanced chemiluminescence immunodetection (Immobilon, USA) was used to visualize immunoreactive proteins. The antibodies for Western blotting were as follows: anti-KIF22 (13403-1-AP; Proteintech Group, USA), anti-Phospho-MEK1/2 (CST: #9154; Proteintech Group), anti-MEK (CST: #8727; Cell Signaling Technology, USA), anti-Phospho-ERK1/2 (CST: #4370; Proteintech Group), anti-ERK (16443-1-AP; Proteintech Group), anti-Cyclin A2 (18202-1-AP; Proteintech Group), anti-Cyclin B1 (55004-1-AP; Proteintech Group), anti-Cyclin D1 (26939-1-AP; Proteintech Group), anti-P21 (10355-1-AP; Proteintech Group), and β-actin (20536-1-AP; Proteintech Group).
3
Protein Extraction and Western Blotting Protocol
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The tissue and cell proteins were isolated using radio immunoprecipitation assay (RIPA) lysis buffur (Epizyme Biotech) supplemented with protease inhibitor. The Bradford method (CWBIO) was used to determine the protein concentrations. Proteins were electrophoresed on Sodium dodecylsulfate-polyacrylamide gel electrophoresis (SDS-PAGE, CWBIO), then transferred to polyvinylidene fluoride (PVDF) membranes, blocked, and incubated with primary anti-FANCE (Biobryt, 1:10000), anti-FANCD2 (1: 5000; Novus Biologicals), anti-GAPDH (1:10000, bioworld), anti-β-tubulin (1:10000, proteintech), anti-RAD51(1:5000, Abcam), anti-γH2AX (1:10000, Abcam), anti-cyclin dependent kinase 4 (CKD4) (1:5000, proteintech), anti-cyclin dependent kinase 6 (CDK6) (1:5000, proteintech), anti-cyclin D1 (1:5000, proteintech), anti-cyclin B1(1:5000, proteintech), anti-cyclin E(1:5000, proteintech) and anti-cyclin A2(1:5000, proteintech) for overnight at 4℃. Secondary antibodies were incubated for 2 h the next day. The protein bands on the membranes were detected using Enhanced Chemiluminescent (ECL, NCM Biotech).
4
Comprehensive Western Blotting Protocol
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Western blotting method was used in the following experiment [14 (link), 15 ], loading 25 μg of total protein lysate per lane. The following primary antibodies were used: anti-HDAC7 (1:1000, #33,418, CST), anti-c-Myc (1:1000, #5605, CST), anti-CDK1 (1:1000, 19,532-1-AP, Proteintech), anti-Cyclin B1 (1:1000, 55,004-1-AP, Proteintech), anti-CDK2 (1:1000, 10,122-1-AP, Proteintech), anti-CyclinA2 (1:1000, 18,202-1-AP, Proteintech), anti-E-cadherin (1:1000,#14,472, CST), anti-Zeb1 (1:1000, 21,544-1-AP, Proteintech), anti-α-SMA (1:1000, 14,395-1-AP, Proteintech) and anti-β-actin (1:1000, #3700, CST). The secondary antibodies used were HRP-linked anti-IgG antibodies (1:5000, Zhongshan Company, Beijing, 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!