Cells were harvested, lysed, and processed for western blot analysis as described previously using 150mM NETN lysis buffer (20 mM Tris [pH 8.0], 150 mM NaCl, 1 mM EDTA, 0.5% NP-40, 1 mM phenylmethylsulfonyl fluoride, 10 mg/mL leupeptin, and 10 mg/mL aprotinin). For cell fractionation, we isolated cytoplasmic and soluble nuclear fractions with the NE-PER Kit (Thermo Scientific) according to the manufacturer’s protocol; to isolate the chromatin fraction, the insoluble pellet was resuspended in RIPA buffer and sonicated in a BioRuptor according to the manufacturer’s protocol (high power, 15 min, 30 s on and 30 s off at 4°C). Proteins were separated using SDSPAGE and electrotransferred to nitrocellulose membranes. Membranes were blocked in 5% milk PBS-Tween and incubated with primary antibody for 1 hr. Abs for western blot analysis included anti-PCNA (Abcam), anti-H2B (Cell Signaling Technology), anti-β-actin (Sigma), anti-FANCJ (E67), anti-HLTF (Abcam), anti-SMARCA1 (Abcam), and anti-HLTF (Santa Cruz Biotechnology). Membranes were washed, incubated with horseradish-peroxidase-linked secondary antibodies (Amersham), and detected by chemiluminescence (Amersham).
Anti hltf
Manufactured by Santa Cruz Biotechnology
Anti-HLTF is a laboratory reagent used for the detection and quantification of HLTF (Helicase-Like Transcription Factor) protein in various biological samples. It is designed to facilitate research and analysis related to the HLTF protein and its role in cellular processes.
Automatically generated - may contain errors
Lab products found in correlation
Anti β actin, by Merck Group (2 mentions)
Horseradish peroxidase linked secondary antibody, by Cytiva (2 mentions)
Anti h2b, by Cell Signaling Technology (2 mentions)
Anti smarca1, by Abcam (2 mentions)
Ne per kit, by Thermo Fisher Scientific (2 mentions)
Anti hltf, by Abcam (2 mentions)
Anti pcna, by Abcam (2 mentions)
2 protocols using anti hltf
1
Cell Fractionation and Western Blot Analysis
2
Cell Fractionation and Western Blot Analysis
Check if the same lab product or an alternative is used in the 5 most similar protocols
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Cells were harvested, lysed, and processed for western blot analysis as described previously using 150mM NETN lysis buffer (20 mM Tris [pH 8.0], 150 mM NaCl, 1 mM EDTA, 0.5% NP-40, 1 mM phenylmethylsulfonyl fluoride, 10 mg/mL leupeptin, and 10 mg/mL aprotinin). For cell fractionation, we isolated cytoplasmic and soluble nuclear fractions with the NE-PER Kit (Thermo Scientific) according to the manufacturer’s protocol; to isolate the chromatin fraction, the insoluble pellet was resuspended in RIPA buffer and sonicated in a BioRuptor according to the manufacturer’s protocol (high power, 15 min, 30 s on and 30 s off at 4°C). Proteins were separated using SDSPAGE and electrotransferred to nitrocellulose membranes. Membranes were blocked in 5% milk PBS-Tween and incubated with primary antibody for 1 hr. Abs for western blot analysis included anti-PCNA (Abcam), anti-H2B (Cell Signaling Technology), anti-β-actin (Sigma), anti-FANCJ (E67), anti-HLTF (Abcam), anti-SMARCA1 (Abcam), and anti-HLTF (Santa Cruz Biotechnology). Membranes were washed, incubated with horseradish-peroxidase-linked secondary antibodies (Amersham), and detected by chemiluminescence (Amersham).
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