Epr4447

Manufactured by Abcam

Sourced in United States

EPR4447 is a mouse monoclonal antibody that recognizes the Endoplasmic Reticulum Oxidoreductin 1-Lα (ERO1L) protein. ERO1L is an endoplasmic reticulum-resident oxidoreductase enzyme involved in the oxidative folding of proteins.

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Lab products found in correlation

Tri methyl histone h3, by Cell Signaling Technology (1 mentions) Histone h3 17168 1 ap, by Proteintech (1 mentions) Gapdh 60004 1 ig, by Proteintech (1 mentions) Puromycin, by Avantor (1 mentions) Aprotinin, by Avantor (1 mentions) Leupeptin, by Avantor (1 mentions) Phenyl methyl sulfonyl fluoride (pmsf), by Avantor (1 mentions) Epr4602, by Abcam (1 mentions) Tetradecanoyl phorbol acetate, by Merck Group (1 mentions) Pepstatin a, by Avantor (1 mentions) Enhanced chemiluminescence detection kit, by Merck Group (1 mentions) Gtx107737, by GeneTex (1 mentions) Epr8830, by Abcam (1 mentions) Ep8589, by Abcam (1 mentions) Reducing agent, by Thermo Fisher Scientific (1 mentions) Trans blot, by Bio-Rad (1 mentions)

4 protocols using epr4447

Comprehensive Histone and Cell Cycle Protein Analysis

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The EZH2 (D2C9) Rabbit mAb (#5246), SUZ12 (D39F6) XP® Rabbit mAb (#3737), PARP (46D11) Rabbit mAb (#9532), Ubiquitin (P4D1) Mouse mAb (#3936), CDK4 (D9G3E) Rabbit mAb (#12790), Phospho-Rb (Ser807/811) (D20B12) Rabbit mAb (#8516), Tri-Methyl-Histone H3 (Lys27) (C36B11) Rabbit mAb (#9733), Tri-Methyl-Histone H3 (Lys4) (C42D8) Rabbit mAb (#9751), Tri-Methyl-Histone H3 (Lys9) (D4W1U) Rabbit mAb (#13969) and Tri-Methyl-Histone H3 (Lys79) (E8B3M) Rabbit mAb (#74073) were obtained from Cell Signaling Technology. Anti-EZH2 antibody (ab283270) and anti-STUB1/CHIP antibody (EPR4447) were obtained from Abcam. Anti-Flag (F2555-100UL) were obtained from Sigma (Danvers, MA, USA). GAPDH(60004-1-Ig), RB(10048-2-Ig) and Histone H3(17168-1-AP) were obtained from Proteintech. All antibodies were diluted according to the instructions and used for western blotting experiments.

Mei H., Wu H., Yang J., Zhou B., Wang A., Hu C., Qi S., Jiang Z., Zou F., Wang B., Liu F., Chen Y., Wang W., Liu J, & Liu Q. (2023). Discovery of IHMT-337 as a potent irreversible EZH2 inhibitor targeting CDK4 transcription for malignancies. Signal Transduction and Targeted Therapy, 8, 18.

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Immunoprecipitation and Immunoblotting Assay

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Antibodies to EBNA1 (sc-57719, Santa cruz), FLAG (M2, Sigma), SUMO1 (Y299, Abcam), SUMO2/3 (for IB/IF, EPR4602, Abcam; for IP, fsc-393144, Santa cruz), STUB1 (EPR4447, Abcam), KAP1 (20C1, Abcam), GST (12G8, M20007, Abmart) and GAPDH (G8140-01, US Biological) were used according to the manufacturers specifications. The monoclonal antibody anti-myc (9E10) and HA (12CA5) were prepared from hybridoma cultures stored in the laboratory. Tetradecanoyl Phorbol Acetate (TPA) was purchased from Sigma and sodium butyrate from J&K Corporation. Proteasome inhibitors PMSF, Leupeptin, Aprotinin, Pepstatin A and Puromycin were purchased from Amresco.

Wang Y., Du S., Zhu C., Wang C., Yu N., Lin Z., Gan J., Guo Y., Huang X., He Y., Robertson E., Qu D., Wei F, & Cai Q. (2020). STUB1 is targeted by the SUMO-interacting motif of EBNA1 to maintain Epstein-Barr Virus latency. PLoS Pathogens, 16(3), e1008447.

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Protein Immunoblotting for FOXP3, STUB1, and Akt

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Total protein was extracted from cell homogenates and then immunoblotted with the following antibodies at 4 °C overnight: mouse mAb against human FOXP3 (236A/E7, Abcam, USA) at 1:200, rabbit pAb against human FOXP3 (GTX107737, GeneTex, USA) at 1:800, rabbit mAb against human STUB1 (EPR4447, Abcam, USA) at 1:500, rabbit mAb against ubiquitin (EPR8830, Abcam, USA)at 1:1,000, rabbit mAb against K48-linked ubiquitin (EP8589, Abcam, USA) at 1:1,000, rabbit mAb against Akt(pan) (#4685, CST, USA) at 1:1,000, rabbit mAb against pAkt(S473) (#4060, CST, USA) at 1:1,000, and rabbit mAb against pAkt(T308) (no.13038, CST, USA) at 1:1,000. After the final wash, membranes were incubated with horseradish peroxidase-conjugated secondary immunoglobin G (IgG) antibody (Jackson ImmunoResearch Laboratories, Inc., PA, USA) (1:10,000) at room temperature for 1 h. Membranes were developed using the Enhanced Chemiluminescence Detection Kit (Millipore, Germany).

Zhang L., Li Q., Xu J., Sun G, & Xu Z. (2020). Cimetidine promotes STUB1-mediated degradation of tumoral FOXP3 by activating PI3K-Akt pathway in gastric cancer. Annals of Translational Medicine, 8(20), 1304.

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Western Blot Analysis of CHIP and Tau

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Reaction samples were taken and quenched with LDS buffer containing reducing agent (Invitrogen) in a 2:1 volumetric ratio (sample:LDS) unless otherwise stated. Proteins were separated on NuPAGE Bolt 4-12% SDS-polyacrylamide gels (Invitrogen) and subsequently scanned on a Typhoon scanner using the indicated excitation and emission modules. A Trans-Blot (Biorad) system was used to transfer proteins separated by SDS-PAGE to PVDF membranes. Primary rabbit monoclonal anti-CHIP (EPR4447, Abcam) or mouse monoclonal anti-tau (1E1/A6, Millipore) were recognized by secondary anti-rabbit or anti-mouse antibodies carrying AlexaFluor-647 labels (Invitrogen) and detected on the Typhoon scanner using the same module as above.

Zuo Y., Chong B.K., Jiang K., Finley D., Klenerman D, & Ye Y. (2020). A General in Vitro Assay for Studying Enzymatic Activities of the Ubiquitin System. Biochemistry, 59(7).

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