Cells of MGC803 and BGC823 treated with formaldehyde, then DNA–protein cross-links were sonicated into fragments of 200-1000 bp and received immunoprecipitate with NFIC-specific antibody and control anti-IgG (Millipore, Bedford, MA) overnight. The precipitated chromatin DNA was retrieved by adding beads and analyzed with RT-qPCR.
Control anti igg
Manufactured by Merck Group
Sourced in United States
Control anti-IgG is a laboratory reagent used to validate and monitor the performance of immunoassay tests that detect immunoglobulin G (IgG) antibodies. It provides a consistent and standardized control sample to ensure the accuracy and reliability of IgG antibody measurement procedures.
Automatically generated - may contain errors
Lab products found in correlation
Ez magna rip kit, by Merck Group (2 mentions)
Anti eif4a3, by Abcam (2 mentions)
Anti ago2, by Merck Group (2 mentions)
Magna rip rna binding protein immunoprecipitation kit, by Merck Group (2 mentions)
Linear ion trap quadrupole mass spectrometer, by Thermo Fisher Scientific (1 mentions)
Pierce rna 3 end desthiobiotinylation kit, by Thermo Fisher Scientific (1 mentions)
Magnetic rna protein pull down kit, by Thermo Fisher Scientific (1 mentions)
Mmessage mmachine t7 transcription kit, by Thermo Fisher Scientific (1 mentions)
Human anti ago2, by Merck Group (1 mentions)
Rna binding protein immunoprecipitation kit, by Merck Group (1 mentions)
9 protocols using control anti igg
1
Chromatin Immunoprecipitation Assay for NFIC
2
Characterization of FAM83H-AS1 RNA Interactome
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RNA immunoprecipitation was performed as described previously,11 and magnetic beads were conjugated with anti‐HNRNPK (ABCAM) or control anti‐IgG (Millipore) antibody. In vitro translation assays were performed using mMESSAGE mMACHINE T7 Transcription Kit (Invitrogen) according to the manufacturer's instructions. Then, FAM83H‐AS1 RNAs were labeled with desthiobiotinylation using the Pierce RNA 3′End Desthiobiotinylation Kit (Thermo Fisher). RNA pull‐down assays were performed with Magnetic RNA‐Protein Pull‐Down Kit (Thermo Fisher) according to the manufacturer's instructions. After elution of RNA‐interacting proteins, they were subjected to mass spectrometric analysis. Liquid chromatography–mass spectrometry experiments were performed with a linear ion trap quadrupole mass spectrometer (Thermo Finnigan) equipped with a micro‐spray source.
3
Analyzing RNA Interaction with Ago2 Protein
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An RNA-binding protein immunoprecipitation (RIP) assay was performed according to the instructions of an EZ-Magna RIP kit (Millipore Corp., Billerica, MA, USA). In brief, cardiomyocytes at an 80–90% confluence were lysed in RIP lysis buffer. Next, the cells lysates were co-incubated with magnetic beads conjugated with human anti-Ago2 (Millipore) or control anti-IgG (Millipore). After that, the samples were incubated with Proteinase K and the immunoprecipitated RNA was collected. Relative abundancy of miR-671 and TGFBR2 was then examined by RT-qPCR.
4
circRNA_100290 RIP Assay Protocol
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RNA-binding protein immunoprecipitation (Millipore, USA) was used to perform an RIP assay. According to the manufacturer’s protocol, 1 × 107 cells were harvested and lysed in complete RIPA lysis buffer. RNA magnetic beads were conjugated to anti-EIF4A3 (Abcam, USA) or control anti-IgG (Millipore, USA). The Ct value of circRNA_100290 was detected by qRT-PCR.
5
Ago2-Associated RNA Interaction Profiling
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RIP assay was undertaken for RNA interaction, following the established protocol of Magna RIP™ RNA-Binding Protein Immunoprecipitation Kit (Millipore, Bedford, MA, USA). Cell lysates were incubated with the anti-Ago2 (1: 200; Millipore) or control anti-IgG (1: 200; Millipore) on 30 μl of magnetic beads at 4 °C overnight. After that, the recovered immunoprecipitates were analyzed using RT-qPCR. This assay was executed in triplicate.
6
Investigating PITPNA-AS1, miR-520d-5p, and SIK2 Interactions
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Based on the manufacturer’s instruments, an EZ-Magna RIP kit (Millipore) was applied. In brief, HCC1937 and MDA-MB-468 cells were washed with pre-cooled PBS and then lysed in RIPA lysis buffer in an ice bath for 5 min, and centrifuged at 12,000×g for 10 min at 4 °C. Then, cell extract was incubated with magnetic beads coated human anti-Ago2 (Millipore), anti-DDX54 (Millipore) or control anti-IgG (Millipore) at 4 °C overnight. Finally, expression of PITPNA-AS1, miR-520d-5p, SIK2 immunoprecipitated by Ago2 and expression of PITPNA-AS1, SIK2 immunoprecipitated by DDX52 were analyzed by RT-qPCR after being purified by proteinase K.
7
Investigating circTNPO3-miR-1299 Interaction
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The interaction between circTNPO3 and miR-1299 was assessed by a Magna RIP RNA-Binding Protein Immunoprecipitation Kit from Millipore (Bedford, MA, USA) referring to manufacturer’s instruction. Briefly, SKOV3/PTX and MeyA-8/PTX cells were lysed in complete RIP lysis buffer on the ice for 5 min. Then the RIP lysate was incubated with magnetic beads conjugated with human anti-Argonaute 2 (AGO2; Millipore) or control anti-IgG (Millipore) antibody in RIP immunoprecipitation buffer for 6 h at 4°C. After incubating with proteinase K, purified RNA was extracted and analyzed for circTNPO3 and miR-1299 expression by qRT-PCR.
8
Ago2-RIP Assay for circRNA and miRNA
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The Ago2‐RIP experiments were performed using an RNA‐Binding Protein Immunoprecipitation Kit (Millipore, USA). Briefly, NP cells were lysed in RNA lysis buffer and then added to the magnetic beads. The magnetic beads were conjugated to human anti‐Ago2 antibody or control anti‐IgG (Millipore) antibody in RIP immunoprecipitation buffer for 6 h at 4°C. Next, the samples were incubated with Proteinase K; immunoprecipitated RNA was then isolated and collected. Furthermore, purified RNAs were detected and analysed by RT‐qPCR to demonstrate the enrichment of circ‐83756 and miR‐558.
9
RIP Assay for circRNA_100290 Binding
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The RNA-Binding Protein Immunoprecipitation (Millipore, USA) was used to perform a RIP assay.
According to the manufacturer's protocol, 1 × 10 7 cells were harvested and lysed in complete RIPA lysis buffer. RNA magnetic beads were conjugated to anti-EIF4A3 (Abcam, USA) or control anti-IgG (Millipore, USA). The Ct value of circRNA_100290 was detected by qRT-PCR.
According to the manufacturer's protocol, 1 × 10 7 cells were harvested and lysed in complete RIPA lysis buffer. RNA magnetic beads were conjugated to anti-EIF4A3 (Abcam, USA) or control anti-IgG (Millipore, USA). The Ct value of circRNA_100290 was detected by qRT-PCR.
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