Ab133565

Manufactured by Abcam

Sourced in China

Ab133565 is a lab equipment product offered by Abcam. It is designed for use in research applications. The core function of this product is to provide a tool for researchers to conduct their experiments.

Automatically generated - may contain errors

Lab products found in correlation

Ab166917, by Abcam (3 mentions) Pierce ecl kit, by Thermo Fisher Scientific (3 mentions) Ab151743, by Abcam (2 mentions) Ab75817, by Abcam (2 mentions) Protein a g magnetic beads, by Selleck Chemicals (1 mentions) Ab11048, by Abcam (1 mentions) Cycloheximide (chx), by Merck Group (1 mentions) Ap51072, by Abcepta (1 mentions) Ab219650, by Abcam (1 mentions) C4859, by Merck Group (1 mentions) Ab2185, by Abcam (1 mentions) Ab92554, by Abcam (1 mentions) Ab184177, by Abcam (1 mentions) Ipsn07852, by Merck Group (1 mentions) Protease inhibitor, by Thermo Fisher Scientific (1 mentions) Ab67035, by Abcam (1 mentions) Ab8245, by Abcam (1 mentions)

4 protocols using ab133565

Immunoprecipitation of Endogenous Proteins

Check if the same lab product or an alternative is used in the 5 most similar protocols

To immunoprecipitate endogenous proteins, cell extracts were incubated with primary antibodies p14/ARF (AB11048, Abcam), RBX1 (ab133565, Abcam) or control IgG in a rotating incubator overnight at 4 °C, followed by incubation with protein A/G magnetic beads (Bimake) for another 2 h. The immunoprecipitates were washed three times with lysis buffer and analyzed by immunoblotting.

Zhang W., Li L., Cai L., Liang Y., Xu J., Liu Y., Zhou L., Ding C., Zhang Y., Zhao H., Qin J., Shao Z., Wei W, & Jia L. (2021). Tumor-associated antigen Prame targets tumor suppressor p14/ARF for degradation as the receptor protein of CRL2Prame complex. Cell Death and Differentiation, 28(6), 1926-1940.

+ Open protocol

+ Expand

Analyzing Ubiquitin Ligase Complex

Check if the same lab product or an alternative is used in the 5 most similar protocols

Cell lysates were generated and analyzed via immunoblotting. We used antibodies against RBX1 (ab133565), Cullin2 (ab166917), and ELOB (ab151743), all sourced from Abcam. During the cycloheximide (CHX) chase assays, the cells were treated with 50 μg/mL CHX (C4859, Sigma, Germany), alongside either 1.0 μM/L MLN4924 or DMSO, for predetermined periods.

Sui X.Y., Ma X.Y., Hou Y., Cao S.W., Wang Z.Q., Jia L.J., Fan L., Shao Z.M, & Zhang W.J. (2024). Elongin B promotes breast cancer progression by ubiquitinating tumor suppressor p14/ARF. Cell Biology and Toxicology, 40(1), 24.

+ Open protocol

+ Expand

Cullin Protein Regulation by CHX-chase

Check if the same lab product or an alternative is used in the 5 most similar protocols

Cell lysates were prepared and analyzed by immunoblotting. Antibody against p14/ARF (AP51072) was purchased from ABGENT (Suzhou city, Jiangsu Province, China). Antibodies against RBX1 (ab133565), Cullin1 (ab75817), Cullin2 (ab166917), Cullin5 (ab18477), Cullin7 (ab96861), EloB (ab151743), Prame (ab219650) were purchased from Abcam (Shanghai, China). Anitbodies againgst Cullin3 (2759), Cullin4A (2699) were from Cell Signaling Technology (Danvers, MA, USA). Antibody against Cullin4B (12916) was from Proteintech (Wuhan, Hubei, China).
For CHX-chase experiments, cells were treated with 50 μg/mL CHX (Sigma, C4859, Merck KGaA, Darmstadt, Hesse, Germany) in combination with 1.0 μmol/L MLN4924 or DMSO for indicated time points [64 (link)].

+ Open protocol

+ Expand

Western Blot Analysis of Cullin Proteins

Check if the same lab product or an alternative is used in the 5 most similar protocols

Western blotting analysis was performed as described previously [16 (link)]. Briefly, total proteins were extracted from biopsies and cultured cells using ice-cold radioimmunoprecipitation assay (RIPA) buffer (#89900, Thermo Fisher) containing 1× protease inhibitor (#87785, Thermo Fisher). Equal amounts of total proteins were separated by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) gels and transferred onto polyvinylidene difluoride membranes (#IPSN07852, Sigma-Aldrich). The membranes were blocked with 5% fat-free milk, followed by probing with the following primary antibodies: anti-CUL1 (#ab75817, Abcam, Shanghai, China), anti-CUL2 (#ab166917, Abcam), anti-CUL3 (#ab108407, Abcam), anti-CUL4A (#ab92554, Abcam), anti-CUL4B (#ab67035, Abcam), anti-CUL5 (#ab184177, Abcam), anti-CUL7 (#C1743, Sigma-Aldrich), anti-CUL9 (#HPA052-004, Sigma-Aldrich), anti-SKP1 (#ab76502, Abcam), anti-RBX1 (#ab133565, Abcam), anti-FBXL1 (SAB1100391, Sigma-Aldrich), anti–hypoxia-inducible factor 1α (HIF1α) (#ab2185, Abcam), anti-MEN1 (#ABC514, Sigma-Aldrich), anti-Myc (#ab32, Abcam), anti-HA (#H3663, Sigma-Aldrich), anti-Flag (#F3165, Sigma-Aldrich), and anti-GAPDH (#ab8245, Abcam). After rinsing five times with PBST buffer, the membranes were probed with secondary antibodies. Finally, protein bands were visualized using the Pierce ECL kit (#32106, Thermo Fisher Scientific).

Zeng J., Xiao X.Q, & Zhou Z.Y. (2021). A Hypoxia-Induced SCFFBXL1 E3 Ligase Ubiquitinates and Degrades the MEN1 Tumor Suppressor to Promote Colorectal Cancer Tumorigenesis. Cancer Research and Treatment : Official Journal of Korean Cancer Association, 54(2), 525-540.

+ 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?

Revolutionizing how scientists
search and build protocols!