Protein samples were mixed with one-third volume of NuPAGE® LDS Sample Buffer (4×) (Life Technologies), boiled at 95°C for 5 min, and then electrophoresed on a 4–12% Bis-Tris gel (Life Technologies). Standard SDS-PAGE and western blotting protocols were carried out afterwards. Primary antibodies used were as follows: anti-hTERT antibody (Abcam, ab32020, 1:1000), anti-β-actin antibody (Sigma, A5441, 1:5000). Secondary antibodies used were as follows: peroxidase-AffiniPure donkey anti-rabbit IgG (H + L) (Jackson, 711-035-152, 1:5000), peroxidase-AffiniPure donkey anti-mouse IgG (H + L) (Jackson, 715-035-150, 1:5000). SuperSignal® West Pico Chemiluminescent Substrate (Thermo Scientific) was used to generate signals on western blots. The signals were detected with a FluorChem HD2 imaging system (Alpha Innotech) and quantified with ImageQuant TL v2005 software.
Anti htert antibody
Manufactured by Abcam
The Anti-hTERT antibody is a research-use only product that specifically recognizes the human telomerase reverse transcriptase (hTERT) protein. hTERT is a critical component of the telomerase enzyme complex, which plays a role in maintaining telomere length and cellular immortalization.
Automatically generated - may contain errors
Lab products found in correlation
Anti β actin antibody, by Merck Group (1 mentions)
Nupage lds sample buffer 4, by Thermo Fisher Scientific (1 mentions)
Bis tris gel, by Thermo Fisher Scientific (1 mentions)
Supersignal west pico chemiluminescent substrate, by Thermo Fisher Scientific (1 mentions)
Fluorchem hd2 imaging system, by Bio-Techne (1 mentions)
Peroxidase affinipure donkey anti rabbit igg h l, by Jackson ImmunoResearch (1 mentions)
Peroxidase affinipure donkey anti mouse igg h l, by Jackson ImmunoResearch (1 mentions)
Annexin 5 fitc kit, by Abcam (1 mentions)
Enhanced chemiluminescence western blotting detection reagent, by GE Healthcare (1 mentions)
Methyl thiazolyl tetrazolium (mtt), by Merck Group (1 mentions)
Rnase a, by Merck Group (1 mentions)
Bibr 1532, by Cayman Chemical (1 mentions)
2 protocols using anti htert antibody
1
Western Blot Analysis of hTERT Expression
2
Endometrial Cancer Cell Line Cultivation
Check if the same lab product or an alternative is used in the 5 most similar protocols
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The endometrial cancer cell lines, ECC-1 and Ishikawa, were used. The ECC-1 cell line was maintained in RPMI 1640 supplemented with 5% FBS, 300mM L-glutamine, 5 μg/ml bovine insulin, 10,000 U/ml penicillin and 10,000 μg/ml streptomycin. Ishikawa cells were grown in MEM supplemented 5% FBS, 300 mM L-glutamine, 10,000 U/ml penicillin and 10,000 μg /ml streptomycin. Both the cell lines were cultured at 37°C in a humidified atmosphere with 5% CO2. MTT (3-(,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) and RNase A were purchased from Sigma (St. Louis, MO). The anti-MMPs and anti-α-tubulin antibodies were purchased from Cell Signaling (Beverly, MA). The anti-hTERT antibody and Annexin V FITC kit were purchased from Abcam (Cambridge, MA). The Enhanced chemiluminescence Western blotting detection reagents were purchased from GE Health care (GE Healthcare Life Sciences, Piscataway, NJ). BIBR1532 was purchased from Cayman (Ann Arbor, MI) and dissolved in DMSO. All other chemicals were purchased from Sigma (St Louis, MO).
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