METHODS: We treated 3T3 - L1 adipocytes with 2.5 mmol / l R (+) alpha - lipoic acid for 2 to 60 min, followed by assays of: 2 - deoxyglucose uptake; glucose transporter 1 and 4 (GLUT1 and GLUT4) subcellular localization; tyrosine phosphorylation of the insulin receptor or of the insulin receptor substrate - 1 in cell lysates; association of phosphatidylinositol 3 - kinase activity with immunoprecipitates of proteins containing phosphotyrosine or of insulin receptor substrate - 1 using a in vitro kinase assay; association of the p85 subunit of phosphatidylinositol 3 - kinase with phosphotyrosine proteins or with insulin receptor substrate - 1; and in vitro activity
of immunoprecipitated Akt1.
Tip 6: For detection
of immunoprecipitated proteins by Western blotting, without or reduced detection of non-specific artifacts (such as the heavy and light chains of the immunoprecipitating antibody), detect primary antibodies using HRP - conjugated anti-rabbit light chain - specific (L) antibody and HRP - conjugated Protein A instead of traditional HRP - conjugated secondary antibodies.
MALDI - MS spectra
of immunoprecipitated Aβ peptides from rat CSF.
Not exact matches
Significant binding by miR -124-3p to two
of the potential target genes was seen, as measured from
immunoprecipitated RNA - induced silencing complexes.
Nagaki et al. [6] used anti-CenH3 antibodies to
immunoprecipitate chromatin (ChIP) comprising DNA bound to CenH3 - containing nucleosomes, confirming that CenH3 is associated with both the CentO repeats and the CRR family
of retrotransposons.
The application
of next - gen sequencing to RNA libraries (RNA - Seq), chromatin
immunoprecipitates (CHiP - Seq), bisulfite - treated DNA, and regions
of open chromatin (DNAse - Seq) makes it possible to interrogate many aspects
of the genome in high - throughput fashion.
Immunoprecipitated samples were resuspended in 10 μl
of TA (33 % acetonitrile, 0.1 % trifluoroacetic acid) and sonicated in a water bath for 10 min.
It will make use
of cutting - edge approaches in vivo and in vitro, including mouse genetic models and next - generation sequencing
of RNA and
immunoprecipitated chromatin (RNA - seq and ChIP - seq).
Protein - DNA complexes were then
immunoprecipitated overnight with polyclonal Oct4 antibody (sc - 9081, Santa Cruz Biotechnology), a mix
of monoclonal Oct4 antibodies (BD Transduction Labs, Santa Cruz Biotechnology, Chemicon), or normal serum controls (Pierce) on a rotator at 4 °C.
HIF -1-alpha was
immunoprecipitated using 0.5 mg HeLa Nuclear DFO treated whole cell extract (ab180880), 5µg
of Rabbit polyclonal to HIF1 alpha and 50µl
of protein G magnetic beads (+).
HIF - 1 alpha was
immunoprecipitated using 0.5 mg HeLa Nuclear DFO treated whole cell extract (ab180880), 5 µg
of Mouse monoclonal to HIF - 1 alpha and 50 µl
of protein G magnetic beads (+).
Lysate
of cross-linked HEK293T cells was
immunoprecipitated by an anti-β-catenin antibody or mouse IgG.