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Pierce c myc tag ip co ip kit

Manufactured by Thermo Fisher Scientific
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The Pierce™ c-Myc-Tag IP/Co-IP Kit is a laboratory equipment product designed for immunoprecipitation (IP) and co-immunoprecipitation (Co-IP) assays. It contains antibodies and reagents necessary to facilitate the purification and analysis of c-Myc-tagged proteins and their interacting partners from cell or tissue lysates.

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Lab products found in correlation

Lipofectamine rnaimax, by Thermo Fisher Scientific (2 mentions) Pierce ha tag ip co ip kit, by Thermo Fisher Scientific (1 mentions) Protease inhibitor cocktail, by Roche (1 mentions) Ltq orbitrap elite mass spectrometer, by Thermo Fisher Scientific (1 mentions)

Close spelling variants of this product

Pierce magnetic c‐Myc‐tag IP/Co‐IP kit C-Myc Tag IP/Co-IP kit Myc-tag Co-IP kit Pierce Co-IP kit IP/Co-IP Kit Co-IP kit Myc Tag C-Myc Kit c

4 protocols using pierce c myc tag ip co ip kit

1

Investigating TSC2 and mTOR Protein Interactions

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Proteins from HeLa cells were extracted in buffer with proteinase and phosphatase inhibitor cocktail as mentioned above. Antibody against endogenous TSC2 or mTOR was crosslinked to Dynabeads™ Protein A by disuccinimidyl suberate (DSS) according to manufacturer’s protocol. Protein extracts were incubated overnight at 4°C with the antibody linked protein A beads. After incubation beads were washed three times with protein extraction buffer, then proteins were eluted by heating the beads in protein sample buffer [60 mM Tris (pH 6.8), 10% glycerol, 100 mM DTT, 0.2% bromophenol blue, 2% SDS] at 70°C for 10 mins, followed by immunoblot. An anti- Rheb antibody was used to detect the interactions between TSC2- Rheb or mTOR- Rheb from different conditions. For pulling down the Myc tagged protein, Pierce™ c-Myc-Tag IP/Co-IP Kit from Thermo Fisher was used according to manufacturer’s instruction. For pulling down Halo tagged protein, the Halo-Trap kit from Chromotek was used based on manufacturer’s instruction.
Yang S., Zhang Y., Ting C.Y., Bettedi L., Kim K., Ghaniam E, & Lilly M. (2020). The Rag GTPase regulates the dynamic behavior of TSC downstream of both amino acid and growth factor restriction. Developmental cell, 55(3), 272-288.e5.
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2

Quantifying DNMT3B4 Inhibition by Caffeic Acid

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Cells overexpressing DNMTΔ3B4 were cultured in 100-mm dishes until reaching 80% confluence. Total protein was extracted using Mammalian protein extraction reagent (M-PER) (Thermo Fisher Scientific, UK). The exogenous DNMT protein tagged with c-Myc was isolated using Pierce c-Myc-Tag IP/Co-IP kit (Thermo Fisher Scientific, UK) according to the manufacturer’s instructions. Inhibition of DNMTΔ3B4 by 25–300 µM caffeic acid was quantified using the DNMT activity assay (Abcam, UK) according to the manufacturer’s instructions.
Sae-Lee C., Barrow T.M., Colicino E., Choi S.H., Rabanal-Ruiz Y., Green D., Korolchuk V.I., Mathers J.C, & Byun H.M. (2022). Genomic targets and selective inhibition of DNA methyltransferase isoforms. Clinical Epigenetics, 14, 103.
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3

Investigating UL44-UBC9 Interaction

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To test the interaction of UL44 with UBC9, 293T cells were transiently transfected with pCMV-HA-UL44 (full length or deletion mutants) and pCMV-Myc-UBC9, or pCMV-Myc-NB80 as negative control. After 48 h cells were harvested and co-IP experiments were performed using either Pierce c-Myc-Tag IP/Co-IP Kit or Pierce HA Tag IP/Co-IP Kit following the manufacturer’s protocol (Thermo Fisher Scientific) supplemented with protease inhibitor cocktail (Roche). The bound proteins were analyzed by western blot using anti-HA and anti-Myc antibodies (Sigma-Aldrich).
Chen J., Li G., He H., Li X., Niu W., Cao D, & Shen A. (2021). Sumoylation of the Carboxy-Terminal of Human Cytomegalovirus DNA Polymerase Processivity Factor UL44 Attenuates Viral DNA Replication. Frontiers in Microbiology, 12, 652719.
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4

Characterization of DHX15 Interactome

Cited 2 times
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HEK293T cells were transfected with a pool of siRNAs targeting DHX15 (Supplemental Table 19) using Lipofectamine RNAiMax (ThermoFisher Scientific) according to manufacturer instructions. Cells were transfected again after 24 hours and harvested at day 4. For overexpression experiments, HEK293T cells were transfected with pCMV6-DHX15 or pCMV6-DHX15 R222G. RNA was extracted after 48 hours with Trizol and libraries prepared as described above. Pull down experiments for mass spectrometry were done using the Pierce c-Myc Tag IP/Co-IP Kit (Thermo Scientific). The MS analysis was performed according to the optimized platform as previously reported74 (link). Proteins in the gel band were in-gel digested by trypsin. Resulting peptides were loaded on a nanoscale reverse-phase column, and eluted by a gradient (~30 min). Eluted peptides were detected by an inline LTQ Orbitrap Elite mass spectrometer (Thermo Scientific). Acquired data were searched against the protein database using the Sequest algorithm and filtered to reduce the protein false discovery rate to below 1%. For binary comparison, statistical analysis was also performed based on the G-test. FDR was derived according to biological replicates and null hypothesis75 (link).
Faber Z.J., Chen X., Gedman A.L., Boggs K., Cheng J., Ma J., Radtke I., Chao J.R., Walsh M.P., Song G., Andersson A.K., Dang J., Dong L., Liu Y., Huether R., Cai Z., Mulder H., Wu G., Edmonson M., Rusch M., Qu C., Li Y., Vadodaria B., Wang J., Hedlund E., Cao X., Yergeau D., Nakitandwe J., Pounds S.B., Shurtleff S., Fulton R.S., Fulton L.L., Easton J., Parganas E., Pui C.H., Rubnitz J.E., Ding L., Mardis E.R., Wilson R.K., Gruber T.A., Mullighan C.G., Schlenk R.F., Paschka P., Döhner K., Döhner H., Bullinger L., Zhang J., Klco J.M, & Downing J.R. (2016). The Genomic Landscape of Core-Binding Factor Acute Myeloid Leukemias. Nature genetics, 48(12), 1551-1556.
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