Sentences with word «immunoprecipitate»

Will immunoprecipitate from in vitro translation assays but not from cell lysates.

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.

MALDI - MS spectra of immunoprecipitated Aβ peptides from rat CSF.

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 (+).

ab32518 (purified) at 1/20 immunoprecipitating IKB alpha in HeLa cell lysate (Lane 1).

Twelve hours after transfection, cells were treated with or without IFNα for another 24 h. Cell lysates (Input) were then immunoprecipitated (IP) with anti — IRF - 9 antibody followed by Western blot (WB) analysis with anti-STAT2 antibody.

Significant binding by miR -124-3p to two of the potential target genes was seen, as measured from immunoprecipitated RNA - induced silencing complexes.

We have identified another family required for RNA editing by immunoprecipitating ORRM1 and performing proteomic analysis in collaboration with the Van Wijk lab.

When an antibody to endogenous SUV39H1 was used for immunoprecipitation, MeCP2 was effectively coimmunoprecipitated; conversely, aHA antibodies to HA - tagged MeCP2 could immunoprecipitate SUV39H1 (Fig. 2G).»

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).

MeCP2 can be immunoprecipitated with antibodies prepared against endogenous SUV39H1; on the other hand, immunoprecipitation of SUB39H1 resulted from aHA antibodies to HA - tagged MeCP2.

Unpurified ab32518 used to immunoprecipitate IKB alpha from human HeLa whole cell lysate.

Aβ peptides were immunoprecipitated from rat CSF with the W0 - 2 antibody and further subjected to MALDI - MS analysis.

For this protocol, samples were lysed in 1 × RIPK2 lysis buffer [50 mM Tris (pH 7.5), 0 mM MgCl2, 1 % Triton X-100, 1 mM dithiothreitol, 1 mM EDTA, 1 mM EGTA, with freshly added 1 mM β - glycerophosphate and protease inhibitors (phenylmethylsulfonyl fluoride and aprotinin)-RSB- and immunoprecipitated overnight using 1.5 μg rabbit anti-RIPK2 antibody.

Furthermore, the 60S splicing complex contains C proteins, and it can be immunoprecipitated with 4F4.

Primer pairs were designed that would amplify sequences scattered along the length of the centromere 8 contig, and these were used to sample the immunoprecipitated DNA using a process called ChIP - PCR, showing that the CenH3 - containing region is approximately 750 kb and does not include the small 2.8 kb cluster of CentO that is separated from the three main arrays.

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.

Unpurified ab32518 used to immunoprecipitate IKB alpha from rat PC12 whole cell lysate.

After extensive washing with immunoprecipitation lysis buffer, the immunoprecipitated proteins were analyzed by immunoblotting using specific antibodies against DDX3 and FLAG epitope (ab1257, Abcam, Cambridge, United Kingdom).

Immunoprecipitated samples were resuspended in 10 μl of TA (33 % acetonitrile, 0.1 % trifluoroacetic acid) and sonicated in a water bath for 10 min.

HuH - 7 cell extracts (lane 3) or FLAG - Sp1 - containing cell extracts (lane 4; 1.5 mg each) were immunoprecipitated with anti-FLAG antibody — conjugated beads and the immunoprecipitates were analyzed by SDS - PAGE followed by immunoblotting with anti-FLAG antibody or anti-DDX3 antibody.

Aβ was immunoprecipitated from 50 μl CSF with 5 μg of the anti-Aβ antibody W0 - 2 (EMI Millipore, USA) coupled to protein - G sepharose beads (GE Healthcare, USA) and incubated overnight at 4 °C.

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.

The next day, protein G sepharose was used to immunoprecipitate the RIPK2 protein complex immunoprecipitation (IP) for 1.5 hours.

The immunoprecipitated chromatin by anti — IRF - 9 antibody was analyzed by PCR using primers specific for RIG - G promoter sequences (+11 to − 288).

Lysate of cross-linked HEK293T cells was immunoprecipitated by an anti-β-catenin antibody or mouse IgG.

Human embryonic kidney cells were transfected with Flag - tagged human Gasp, and lysates from these cells were immunoprecipitated with anti-Flag antibody.

Toward this goal, CKD9 was immunoprecipitated from the nuclear extracts of T47D control and LARP7 KD cells, and its associated factors were analyzed by Western blotting.

After 24 h, cells were incubated with or without PP2, lysed, and immunoprecipitated with anti-HA (left) or anti-myc (right) or control Abs (ctr Ab) followed by Western blotting with the indicated Abs.

Two different techniques were applied on the same subjects used for microarray experiments to validate the regions called differentially methylated from the microarray analysis (n = 8 CPA and n = 12 Controls); quantitative real - time PCR on the immunoprecipitated DNA samples (Q - MeDIP) and pyrosequencing of bisulfite treated DNA (see File S1 for a more detailed description and Table S4 for the list of primers used).