Tumor and adjacent normal tissues in cohort 2 were used for immunohistochemistry (IHC) analysis. A standard protocol using anti-UBR5 antibody (ab70311; Abcam, Cambridge, MA, USA) was used to detect UBR5 expression. Sections were counterstained with hematoxylin, and immunoreactivity was scored using the H-score system by two investigators on the basis of the proportion of positively stained cells. Patients with ≥ 50% positive cells were graded as highly expressive, whereas those with < 50% positive cells were classified as lowly expressive.
Ab70311
Manufactured by Abcam
Sourced in United States
Ab70311 is a laboratory equipment product manufactured by Abcam. The core function of this product is to provide a tool for research purposes, but a detailed description cannot be provided while maintaining an unbiased and factual approach.
Automatically generated - may contain errors
Lab products found in correlation
Ab39688, by Abcam (1 mentions)
Ab55906, by Abcam (1 mentions)
Ab137852, by Abcam (1 mentions)
Ab101562, by Abcam (1 mentions)
Ab76424, by Abcam (1 mentions)
Ab1791, by Abcam (1 mentions)
A0216, by Beyotime (1 mentions)
Enhanced chemiluminescence detection kit, by Thermo Fisher Scientific (1 mentions)
A0208, by Beyotime (1 mentions)
Ab32572, by Abcam (1 mentions)
Bicinchoninic acid protein assay kit, by Beyotime (1 mentions)
Polyvinylidene fluoride membrane, by Merck Group (1 mentions)
Radioimmunoprecipitation assay lysis buffer, by Beyotime (1 mentions)
Ab52894, by Abcam (1 mentions)
Ab32077, by Abcam (1 mentions)
Ab7780, by Abcam (1 mentions)
Sc 2027, by Santa Cruz Biotechnology (1 mentions)
Protein a g plus agarose bead, by Santa Cruz Biotechnology (1 mentions)
Af5006, by Affinity Biosciences (1 mentions)
4 protocols using ab70311
1
Immunohistochemical Analysis of UBR5 Expression
2
Immunoblotting Analysis of Protein Expression
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Whole protein was extracted using radioimmunoprecipitation assay lysis buffer (Beyotime) and quantified using the bicinchoninic acid Protein Assay Kit (Beyotime). Briefly, a 20 μg protein aliquot was loaded into 10% sodium dodecyl sulfate polyacrylamide gel electrophoresis for separation, followed by electroblotting on a polyvinylidene fluoride membrane (Merck Millipore, Burlington, MA, USA), blocking using 5% fat-free milk, and overnight incubation with the following primary antibodies: anti-UBR5 (ab70311; Abcam), anti-AXIN1 (ab55906; Abcam), anti-β-catenin (ab32572; Abcam), anti-Survivin (ab76424; Abcam), anti-C-myc (ab39688; Abcam), anti-lactate dehydrogenase A (LDHA, ab101562; Abcam), anti-pyruvate kinase isozymes M2 (PKM2, ab137852; Abcam), anti-H3 (ab1791; Abcam), and anti-GAPDH antibody (60004-1-1G; ProteinTech, Rosemont, IL, USA). They were washed thrice with Tris-buffered saline with Tween 20 and incubated with horseradish peroxidase-conjugated secondary antibodies (A0208, A0216; Beyotime). Finally, the blot was imaged using the Enhanced Chemiluminescence Detection Kit (Pierce Biotechnology), and the band intensity was measured using Image-Pro Plus 6.0 software, with GAPDH as the loading control.
3
Immunoprecipitation and Western Blotting
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Cells were lysed with radioimmunoprecipitation assay (RIPA) buffer and incubated with anti‐USP14 (ab70311; Abcam), anti‐EGFR (ab52894; Abcam), or normal IgG (sc‐2027; Santa Cruz Biotechnology, Inc.) antibodies and then incubated with protein A/G PLUS‐Agarose beads (sc‐2003; Santa Cruz Biotechnology, Inc.) for 2 h at 4°C. The immune complex was washed thrice with lysis buffer. Proteins were then detected with standard Western blot analysis using anti‐USP14 (ab175810; Abcam), anti‐EGFR (ab32077; Abcam), and anti‐ubiquitin (ab7780; Abcam) antibodies.
4
Generation of Stable Cell Line Expressing T7 Polymerase
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BSR-T7/5 cells stably expressing the T7 phage RNA polymerase were generated by Buchholz et al. [90 (link)], and were a kind gift from Prof. Xiufan Liu (Key Laboratory of Animal Infectious Diseases, Yangzhou University, China). The parental NDV (rSS1GFP) and the mutant NDV (rSS1GFP-M/NLSm) carrying M/NLS mutation were generated in our previous study [10 (link)], which were plaque purified three times in chicken embryonic fibroblasts and propagated once in specific pathogen-free (SPF) embryonated chicken eggs. The rabbit polyclonal antibodies against NDV M protein and NP protein was prepared in our laboratory. The rabbit polyclonal antibodies against ENY2 (DF12979), BRD2 (DF12857), RPL18 (DF3700), RPL34 (DF3708), PSMD3 (DF3645), RAB12 (DF12459), TRAF6 (AF5376), NF-κB p65 (AF5006), phospho-NF-κB p65 (Ser536) (AF2006), GAPDH (AF7021), GFP (T0006), HA (T0050) and IL-2 (AF5105) were purchased from Affinity Biosciences (USA). The rabbit polyclonal antibodies against POP1 (ab254978), SWD2 (ab220240), UBR5 (ab70311), STX5 (ab217130), and rabbit monoclonal antibody against phospho-TIFA (Thr9) (ab214815) were purchased from Abcam (USA). The rabbit polyclonal antibody against TIFA (61358S) was purchased from Cell Signaling Technology (USA).
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