For immunohistochemical staining (IHC), Anti-CD133 (ZSBIO, Beijing, China) dilution was 1:100. Anti-CD44v6 (ZSBIO) dilution was 1:200. Anti-Cyclin D1 (ZSBIO) dilutions were 1:50. Anti-MRP (ZSBIO) dilutions were 1:50. Anti-P-gp (ZSBIO) dilutions were 1:50. Anti-TERT (Abcam, Cambridge, MA, USA) dilutions was 1:200. Anti-β-catenin (Fuzhou Maixin Biotech. Co., Ltd., Fujian, China) dilutions were 1:1. Anti-E-cadherin (ZSBIO) dilutions was 1:70. Anti-EpCAM (Cell Signaling Technology, Danvers, MA, USA) dilutions was 1:800, and incubated overnight at 4°C. Chromogenic detection was then done using a peroxidase-conjugated secondary antibody and DAB reagents (ZSBIO). The proliferation index was analyzed using the Cyclin D1 antibody. Staining was measured as the percentage of positively stained nuclei in 200 tumor cells in a consecutive field. Counting was conducted on the most intensely stained areas. The immunostains were scored by two pathologists (FL and SGW) blinded to the clinical data.
Anti tert
Manufactured by Abcam
Sourced in United Kingdom
Anti-TERT is a primary antibody that specifically recognizes the Telomerase Reverse Transcriptase (TERT) protein. TERT is a key component of the telomerase complex, which plays a crucial role in maintaining telomere length and cellular immortalization. This antibody can be used to detect and study the expression and distribution of TERT in various cell and tissue samples.
Automatically generated - may contain errors
Lab products found in correlation
Anti e cadherin, by ZSGB-BIO (1 mentions)
Dab reagent, by ZSGB-BIO (1 mentions)
Anti epcam, by Cell Signaling Technology (1 mentions)
Dynabeads protein g, by Thermo Fisher Scientific (1 mentions)
Anti collagen 1, by Abcam (1 mentions)
Anti fn, by Abcam (1 mentions)
Anti trf1, by Abcam (1 mentions)
Supersignal ecl system, by Thermo Fisher Scientific (1 mentions)
Anti sirt6, by Abcam (1 mentions)
Bca protein assay kit, by Beyotime (1 mentions)
β actin, by Abcam (1 mentions)
Radioimmunoprecipitation assay buffer, by Beyotime (1 mentions)
Anti α tubulin, by Abcam (1 mentions)
Anti sirt6, by Cell Signaling Technology (1 mentions)
Anti β tubulin, by Abcam (1 mentions)
Anti sirt3, by Cell Signaling Technology (1 mentions)
Anti creb, by Abcam (1 mentions)
Anti acetylated lysine, by Cell Signaling Technology (1 mentions)
Anti mortalin, by Merck Group (1 mentions)
Anti sirt1, by Santa Cruz Biotechnology (1 mentions)
5 protocols using anti tert
1
Immunohistochemical Profiling of Tumor Markers
2
Immunoprecipitation of TERT Protein
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Cells were lysed with RIPA buffer (1% NP-40 and 0.25% deoxycholate, Beyotime P0013D) containing protease and phosphatase inhibitors and PMSF, and centrifuged at 12,000 ×g for 15 minutes to collect the supernatant. 2,000 μg total protein in 0.5 mL was incubated with anti-TERT (Abcam) antibody overnight at 4°C with rotation. A sample incubated with IgG was included as control. Dynabeads protein G (Thermo Fisher Scientific-Invitrogen) were added to the antibody conjugated complex and incubated for 2 hours at 4°C with rotation. Afterward, the beads were washed three times with washing buffer A (50 mM Tris-HCl, pH 7.5, 0.5% Triton X-100, 150 mM NaCl, 2 mM CaCl2, 5% glycerol, 2 mM PMSF), once with washing buffer B (50 mM Tris-HCl, pH 7.5, 0.5% Triton X-100, 500 mM NaCl, 2 mM CaCl2, 5% glycerol, 2 mM PMSF), and twice with washing buffer C (50mM Tris-HCl, pH 7.5, 150 mM NaCl, 2 mM CaCl2). The bound materials were eluted by boiling in SDS protein sample buffer, and analyzed by western blotting.
3
Western Blot Analysis of Myocardial Proteins
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For western blot analysis, mouse myocardial tissues from infarcted area of TAC groups and anterior left ventricular of sham operated groups were homogenized in radio-immunoprecipitation assay buffer (Beyotime, Shanghai, China) and protein concentration was determined using the BCA protein assay kit (Beyotime). Aliquots containing 20 μg of protein were separated by 10% sodium dodecyl sulfatepolyacrylamide gel electrophoresis and transferred to nitrocelullose membranes (Pall Corporation, USA). Membranes were incubated in 5% nonfat milk in Tris-buffered saline with Tween 20 (TBST) for 1 h at room temperature, followed by incubation in primary antibodies at 4°C overnight. The primary antibodies used and their dilutions were as follows: anti-SIRT6, 1:2,000; anti-α-tubulin, 1:2,000; anti-TERT, 1:1,000; anti-TRF1, 1:1,000; anti-collagen I, 1:5,000; anti-FN, 1:1,000; anti-α-tubulin, 1:1,000; β-actin, 1:2,000 (all from Abcam, Cambridge, United Kingdom). After washing in TBST, membranes were incubated with the appropriate horseradish peroxidase-conjugated secondary antibodies at room temperature for 1 h. Signal detection was performed using the SuperSignal ECL system (ThermoScientific, Waltham, Massachusetts) and bands were analyzed by ImageJ software. Band intensity was normalized to that of α-tubulin or β-actin.
4
Protein Extraction and Western Blotting
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Cells were washed twice in 1× PBS, pH 7.4, and lysed in buffer A (10 mmol/L HEPES, pH 7.9, 10 mmol/L KCl, 1.5 mmol/L MgCl2 and 0.5% NP-40). Mitochondria were lysed directly in 1× SDS loading buffer. Protein lysates (50 μg) were resolved by SDS-PAGE, transferred to nitrocellulose membranes, incubated for 1 h with 5% milk TBS-T and overnight with primary antibodies in 5% BSA at 4 °C or for 1–2 h at room temperature. Antibodies included anti-PNPASE (1:5,000) (Chen et al., 2006 (link)), anti-TIM23 (1:1000) (Abgent), anti-Mortalin (1:10,000) (Sigma-Aldrich), anti-Creb (1:1000) (Abcam), anti-ActB (1:2,000) (ABClonal), anti-Acetylated lysine (1:1000) (Cell Signaling Technology), anti-TERT (1:1000) (Abcam), anti-p-AMPK (Thr172) (1:1000) (Cell Signaling Technology), anti-SIRT1 (1:500) (Santa Cruz Biotechnology), anti-SIRT6 (1:1000) (Cell Signaling Technology), anti-SIRT3 (Cell Signaling Technology), anti-p-NF-kB(Ser536) (1:1000) (Cell Signaling Technology), anti-p16 (1:1000) (Bioworld) and anti-β-Tubulin (1:2000) (Abcam).
5
Protein Extraction and Western Blot Analysis
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Total proteins were extracted as previously described (Maggisano et al., 2014) . Twenty-five mg of total protein extracts were run on a 9% or 12% SDS-PAGE gel and transferred to PVDF membrane (VWR, Milan, Italy), blocked with TTBS/milk (TBS, 1% Tween 20 and 5% non-fat dry milk) and incubated overnight with affinity-purified anti-TERT (Abcam, Cambridge, United Kingdom) and anti-GAPDH antibodies (Thermo Fisher Scientific Inc.), diluted 1:1000 and 1:50,000, respectively. The membranes were incubated with horseradish peroxidase-conjugated anti-rabbit or anti-mouse antibody (Transduction Laboratories, Lexington, KY, USA) in TTBS/ milk, diluted 1:5000 or 1:50,000 respectively. Western blot detection system ECL Plus (Perkin Elmer, Monza, Italy) was used to visualize the proteins.
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