Total proteins were extracted from mouse livers or cultured cells in RIPA buffer containing proteinase inhibitors (Sigma). Nuclear fractions were isolated using NE‐PER™ Nuclear and Cytoplasmic Extraction Reagents (ThermoFisher Scientific, Waltham, MA). Nuclear fractions (10 μg) or total protein lysates (50 μg) were boiled in 1× Laemmli buffer containing 5% β‐mercaptoethanol, separated on 4%–12% Bis‐Tris protein gels (Novex, Carlsbad, CA), and electro‐transferred onto polyvinylidene difluoride membrane for immunoblotting. Primary antibodies used were mouse anti‐Aldh1a1 (Santa Cruz Technologies), rabbit anti‐Yap and anti‐phosphorylated Yap at serine 127 residue (p‐YAP) (Cell Signaling), rabbit anti‐Adh1 (Santa Cruz Technologies), mouse anti‐FLAG antibody (sigma), mouse anti‐Cyclin D1 (Santa Cruz Technologies), rabbit anti‐Hif1α (ThermoFisher Scientific), rabbit anti‐fibronectin (Fn1) (Abcam), rabbit anti‐Vegfa, rabbit anti‐CD11b (Abcam), mouse amti‐Aldh1a1 (Cell Signaling), and Dhrs3 (Proteintech, Rosemont, IL), mouse anti‐Actin (Abcam), and rabbit anti‐GAPDH (Abcam). Detection was carried out using horseradish peroxidase‐conjugated secondary antibodies (Santa Cruz biotechnologies) and the ECL Plus kit (Amersham Biosciences, Piscataway, NJ).
Rabbit anti hif1α
Manufactured by Thermo Fisher Scientific
Rabbit anti-Hif1α is a polyclonal antibody raised against the Hypoxia-Inducible Factor 1-alpha (Hif1α) protein. Hif1α is a transcription factor that plays a key role in the cellular response to hypoxia. The antibody can be used to detect and quantify Hif1α expression in various research applications.
Automatically generated - may contain errors
Lab products found in correlation
Ne per nuclear and cytoplasmic extraction reagent, by Thermo Fisher Scientific (4 mentions)
Proteinase inhibitor, by Merck Group (2 mentions)
Mouse anti flag antibody, by Merck Group (2 mentions)
Ecl plus kit, by Cytiva (2 mentions)
Rabbit anti vegfa, by Abcam (2 mentions)
Rabbit anti cd11b, by Abcam (2 mentions)
Rabbit anti yap and anti phosphorylated yap at serine 127 residue p yap, by Cell Signaling Technology (2 mentions)
Rabbit anti fibronectin fn1, by Abcam (2 mentions)
Dhrs3, by Proteintech (2 mentions)
Bis tris protein gel, by Thermo Fisher Scientific (2 mentions)
Rabbit anti gapdh, by Abcam (2 mentions)
Mouse anti actin, by Abcam (2 mentions)
Horseradish peroxidase conjugated secondary antibody, by Santa Cruz Biotechnology (1 mentions)
Mouse anti α tubulin, by Merck Group (1 mentions)
3 protocols using rabbit anti hif1α
1
Liver and Cell Protein Extraction and Analysis
2
Immunoblotting Analysis of Liver and Cell Proteins
3
Hypoxia Induction in Myoblasts
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Western blot experiments were undertaken as previously described 22 using human myoblasts expanded for 48 hours either in GM (control condition), in GM containing 250 μM of Co2+ or in GM containing 250 μM of deferoxamine (DFO, Sigma-Aldrich). After the procedure, blots were incubated overnight at 4°C with rabbit anti-HIF-1α (1/1000, Thermo Fisher Scientific) or mouse anti-α-tubulin (1/10 000, Sigma-Aldrich) antibodies diluted in TTBS-5% milk. After washing, blots were then incubated with horseradish peroxidase-conjugated goat antimouse or goat anti-rabbit antibodies (1/6000, BioRad, Hercules, CA).
ImageJ software was used to quantify the level of protein expression.
ImageJ software was used to quantify the level of protein expression.
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