Tissues were homogenized and cells were lysed in RIPA buffer and quantified by the bicinchoninic acid assay kit (Pierce, USA). Total protein was separated by sodium dodecyl-sulfate polyacrylamide gel electrophoresis under denaturing conditions and transferred to polyvinylidene fluoride membranes. The membranes were blocked in 5% non-fat milk for 1 h and then incubated with primary antibodies overnight. On the next day, the membranes were incubated with secondary antibodies. Proteins were detected using enhanced chemiluminescence substrate (CST, USA). Antibodies used for western blot are listed below: CDCA5 (ab192237) was purchased from Abcam (UK). AKT (#4685), p-AKT (Ser473, #4060), NF-κB (#8242), Caspase-3 (#9662), and Cleaved caspase-3 (#9665) were purchased from CST (USA).
Cdca5 ab192237
Manufactured by Abcam
Sourced in United Kingdom
CDCA5 (ab192237) is a protein that plays a role in cell division. It is a component of the kinetochore and is involved in the regulation of chromosome segregation during mitosis. The product is suitable for use in Western blot, immunohistochemistry, and immunofluorescence applications.
Automatically generated - may contain errors
2 protocols using cdca5 ab192237
1
Western Blot Analysis of Signaling Proteins
2
Protein Expression Analysis Protocol
Check if the same lab product or an alternative is used in the 5 most similar protocols
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Tissues or tumor cells were lysed with 2% sodium dodecyl sulfate (SDS) and the protein concentration was then measured with BCA protein assay kit. Next, proteins were separated by SDS-PAGE gel and transferred to a nitrocellulose membrane. After blocking, the membrane was incubated with different primary antibodies and secondary antibody (with washing between steps). Finally, the target protein bands were visualized via chemiluminescence system. Primary antibodies include: CDCA5 (ab192237) and eEF1A1 (ab157455) obtained from Abcam; p-AKT (Ser473) antibody (9271), p-S6 (Ser240/244) antibody (5364) and p-4EBP1 (Thr37/46) antibody (2855) were purchased from Cell Signaling Technology; β-actin (sc-69879) and GAPDH (sc-32233) were obtained from Santa Cruz. Secondary antibody Rabbit IgG (#7074) and Mouse IgG (#7076) were bought from Cell Signaling Technology.
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