Sc 271289

Manufactured by Santa Cruz Biotechnology

Sourced in Sweden

Sc-271289 is a laboratory product offered by Santa Cruz Biotechnology. It is a piece of equipment designed for scientific research and analysis. The core function of this product is to facilitate the collection and processing of biological samples. No further details are available regarding its intended use or specific applications.

Automatically generated - may contain errors

Lab products found in correlation

Spelling variants of this product

Anti-Ubiquitin (sc-271289 (2 citations)

4 protocols using sc 271289

Protein Expression Analysis in Cells and Tumors

Cited 1 time

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

Rabbit monoclonal Abs against p-Akt (#4060), total forms of Akt (#2938), p-mTOR (#5536), mTOR (#2972), p-4EBP1 (#9451), 4E-BP1 (#9452), PAK1 (#2602), HA tag (#3714), Atg14 (#96752), Vps34 (#4263), PI3K (#4249), LC3A/B (#4108), Beclin1 (#3495), and Atg5 (#12994) were purchased from Cell Signaling Technology. Mouse monoclonal Abs against ubiquitin (sc-271289) were purchased from Santa Cruz Biotechnology.
The samples derived from cells and tumor homogenates were lysed in Mammalian Protein Extraction Reagent (Thermo Fisher Scientific), separated by electrophoresis on 12% SDS-polyacrylamide gel electrophoresis and transferred to nitrocellulose transfer membranes (Merck Millipore, Cork, Ireland). Proteins were detected using primary Abs at a concentration of 1:1,000 (Cell Signaling Technology) and were incubated overnight. Specific interaction with the primary Abs was detected using corresponding secondary Abs conjugated to HRP (Biosharp), and signals were developed using the enhanced chemiluminescence (ECL) reagents (Biosharp). Gel bands were quantified by ImageJ software, and data are presented as mean ± SD from three independent immunoblotting assays.⁵¹ (link) Phosphorylated and total protein levels were determined and quantified by three successive immunoblotting membranes.

Bu H., Tan S., Yuan B., Huang X., Jiang J., Wu Y., Jiang J, & Li R. (2020). Therapeutic potential of IBP as an autophagy inducer for treating lung cancer via blocking PAK1/Akt/mTOR signaling. Molecular Therapy Oncolytics, 20, 82-93.

+ Open protocol

+ Expand

Immunoprecipitation of Flag, HA, and Ubiquitin

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

Cells were cultured in 6-well plates at a density of 5 × 10⁵ cells/well overnight before being transfected with the indicated plasmids. After 36 h, the cells were incubated with 20 μM MG132 for 4 h. Total proteins were extracted using cell lysis buffer containing a protease/phosphatase inhibitor cocktail (Cell Signaling Technology). Cell lysates were incubated with anti-Flag M2 affinity gel (A2220, Sigma-Aldrich) at 4°C overnight. After washing the beads three times with cell lysis buffer, the immunoprecipitates were collected for immunoblotting. For HA-tagged protein and Ubiquitin protein immunoprecipitation, the lysates were incubated with 1 mg of an anti-HA (3,724, Cell Signaling Technology) antibody, an anti-ubiquitin antibody (sc-271289, Santa Cruz) or IgG overnight and were then incubated with 30 μL of 50% GammaBind plus Sepharose beads (17-0886-01, GE Healthcare, Sweden) for an additional 4 h. After washing the beads three times with cold lysis buffer, the immunoprecipitated proteins were analyzed using immunoblotting.

Fu Y., Zheng Q., Mao Y., Jiang X., Chen X., Liu P., Lv B., Huang T., Yang J., Cheng Y., Dai X., Dai C., Wang X., Yin Y., Song T., Jin W., Zou C., Chen T., Fu L, & Chen Z. (2020). WNT2-Mediated FZD2 Stabilization Regulates Esophageal Cancer Metastasis via STAT3 Signaling. Frontiers in Oncology, 10, 1168.

+ Open protocol

+ Expand

Antibody-Based Protein Detection Assay

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

Monoclonal anti-HA (1:10000, M180–3), anti-Myc (1:10000, M192–3), and anti-Flag (1:10000, M185–3L) were obtained from MBL Beijing Biotech Co., Ltd. (Beijing, China). Antibodies against USP5 (1:1000, 10473–1-AP) and GAPDH (1:5000, 60004–1-Ig) were purchased from Proteintech Group, Inc. (Wuhan, Hubei, China). The anti-c-Maf antibody was obtained from Santa Cruz Biotechnology, Inc. (1:1000, sc-7866) (Santa Cruz, CA, USA) and Proteintech (1:1000, 55013–1-AP). An anti-Ub antibody was obtained from Santa Cruz Biotechnology, Inc. (1:1000, sc-271289). The US FDA-approved drug library and mebendazole were purchased from TargetMol Corp. (Boston, MA, USA). HRP-labeled goat anti-mouse and goat anti-rabbit IgG (H + L) antibodies were purchased from Beyotime Institute of Biotechnology (Nantong, China).

Chen X.H., Xu Y.J., Wang X.G., Lin P., Cao B.Y., Zeng Y.Y., Wang Q., Zhang Z.B., Mao X.L, & Zhang T. (2019). Mebendazole elicits potent antimyeloma activity by inhibiting the USP5/c-Maf axis. Acta Pharmacologica Sinica, 40(12), 1568-1577.

+ Open protocol

+ Expand

Quantitative Western Blot Analysis

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

Western blotting was performed as previously described (Du et al., 2017b; Sun et al., 2017) . Total liver (~30 mg) protein was extracted using a protein extraction kit (C510003; Sangon Biotech Co. Ltd., Shanghai, China). Total protein concentration was estimated by the bicinchoninic acid method (P1511; Applygen Technologies). A total of 30 μg of protein from each sample was separated by 12% SDS-PAGE and electrophoretically transferred to a polyvinylidene difluoride membrane. The membranes were blocked in 3% BSA/ Tris-buffered saline/Tween (TBS-T) buffer for 4 h. The blocked membranes were incubated overnight at 4°C with primary antibodies against p62 (1:2,000; ab101266; Abcam), LC3 (1:1,000; ab48394; Abcam), β-actin (1:2,000; ab8226; Abcam), or ubiquitin (1:200; sc-271289; Santa Cruz Biotechnology, Santa Cruz, CA). The membranes were then washed with TBS-T and incubated with horseradish peroxidase-conjugated antirabbit or antimouse immunoglobulin at room temperature for 45 min. Immunoreactive bands were visualized by enhanced chemiluminescence solution (ECL, Millipore, Bedford, MA). β-Actin was used a reference protein in this study, and each band was related to β-actin. All bands were analyzed using Image-Pro Plus (Media Cybernetics, Rockville, MD).

Du X., Liu G., Loor J.J., Fang Z., Bucktrout R., Yang Y., Ye Q., Shi Z., Shen T., Wang X., Peng Z., Zhao C., Lv B., Xing D., Zhu Y., Li X, & Li X. (2018). Impaired hepatic autophagic activity in dairy cows with severe fatty liver is associated with inflammation and reduced liver function. Journal of dairy science, 101(12).

+ 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!