miR-506 mimics or miRNA was transfected into HCCC-9810 and CCLP1 cells. And the cells were lysed and collected in a RIP lysis buffer kit (Millipore, USA). Human anti-Ago2 (Millipore, USA) or mouse anti-IgG (Millipore, USA) were then conjugated with RNAs magnetic beads. Then the expression levels of purified RNA were determined by qRT-PCR.
Human anti ago2
Manufactured by Merck Group
Sourced in United States
Human anti-Ago2 is a laboratory reagent that targets the Argonaute 2 (Ago2) protein, a key component of the RNA-induced silencing complex (RISC). Ago2 plays a crucial role in the RNA interference (RNAi) pathway, which is involved in gene regulation and post-transcriptional gene silencing. This product can be used in various research applications to study the Ago2 protein and its interactions within the RNAi system.
Automatically generated - may contain errors
Lab products found in correlation
Normal mouse anti igg, by Merck Group (3 mentions)
Rip lysis buffer kit, by Merck Group (2 mentions)
Anti igg mouse, by Merck Group (2 mentions)
Ez magna rip kit, by Merck Group (2 mentions)
Rip buffer, by Merck Group (1 mentions)
Control anti igg, by Merck Group (1 mentions)
Magna rna immunoprecipitation kit, by Merck Group (1 mentions)
Magna rip rna binding protein rip kit, by Merck Group (1 mentions)
Dual luciferase reporter assay system, by Promega (1 mentions)
10 protocols using human anti ago2
1
Investigating miR-506 Regulation by RIP
2
RNA Immunoprecipitation in Cancer Cells
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
T47D and MB231 cells were lysed using RIP buffer (Millipore), and then incubated with magnetic beads coated with human Anti-Ago2 (Millipore) or normal mouse Anti-IgG (Millipore). After incubation with Proteinase K, the immunoprecipitated RNA was isolated and subjected to qRT-PCR analysis as described above.
3
RIP Assay for miRNA Identification
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The RIP assay was performed using the EZ-Magna RiP Kit (Millipore, Billerica, MA, USA). Cells were lysed in lysis buffer and further incubated with magnetic beads together with human anti-Ago2 (Millipore) or normal human IgG control (Millipore). The IP RNAs were extracted with TRIzol and assessed by qRT-PCR.
4
Analyzing RNA Interaction with Ago2 Protein
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
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.
5
RNA-Protein Interaction Profiling via RIP
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Magna RNA immunoprecipitation kit (Millipore, Billerica, MA, USA) was used to perform RIP assay. OE19 cells were lysed using RIP buffer, followed by incubation with magnetic beads coated with magnetic beads conjugated with human anti-Ago2 (Millipore) or normal mouse anti-IgG (Millipore). Following interaction with Proteinase K to digest proteins, the immunoprecipitated RNA was extracted and subjected to qRT-PCR analysis.
6
Quantifying AGO2-bound RNAs by RIP-qPCR
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The Magna RIP RNA-Binding Protein RIP kit (Millipore, Bedford, MA, USA) was used in accordance with the supplied instructions. Briefly, lysates of 2 × 107/l cells were incubated with magnetic beads conjugated with normal mouse IgG (control) or human anti-AGO2 (Millipore, Bedford, MA, USA). Briefly, lysates of 2 × 107cells per liter were incubated with magnetic beads conjugated with 5 µg normal rabbit IgG (cat. #PP64B, Millipore, USA) or 5 µg rabbit anti-AGO2 (cat. #T56652, Abmar, China). The immunoprecipitated RNAs were isolated and verified using qRT-PCR.
7
Ago2 Immunoprecipitation and RNA Analysis
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
16HBE cells were lysed using RIP buffer, then the lysate was incubated with magnetic beads conjugated with human anti-Ago2 (Millipore) or normal mouse anti-IgG (Millipore). After interaction with proteinase K, the immunoprecipitated RNA was eluted and purified RNA was tested using qRT-PCR assay.
8
Verifying RNA-Protein Interactions by RIP
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The RNA immunoprecipitation (RIP) assay was performed to verify the binding of ZEB1-AS1, miR-217 and MAFB. The HK-2 cells, transfected with miR-217 or miR-NC, were lysed in the RIP buffer, and the cell lysis solution was incubated with magnetic beads conjugated with human anti-Ago2 (Millipore) or the negative control mouse IgG antibody (CST, USA). Subsequently, the enrichment of ZEB1-AS1 or MAFB was quantified by qRT-PCR.
9
Investigating PVT1 and miR-145-5p Interaction in RISC Complex
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
RIP assay was performed to explore whether PVT1 and miR-145-5p are together in the same RISC complex. In brief, THP-1 cells were maintained with RIPA lysis buffer to obtain cell lysates. Cell lysates were then incubated with human anti-Ago2 or anti-IgG (Millipore) overnight at 4°C. Then, the RNA-protein complexes were immunoprecipitated with protein A agarose beads. Next, the RNAs were isolated by using TRIzol reagent. Finally, the expression levels of miR-145-5p and PVT1 were measured by qRT-PCR assay. IgG served as the negative control.
10
Luciferase Assay and RNA Immunoprecipitation for miR-506 Interactions
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The sequences containing the wild-type (WT) or mutated (MUT) region DDX5 and linc00473 were synthesized by GenePharma (Shanghai, China) and inserted into a pmirGLO-Report luciferase vector. For the luciferase reporter assay, miR-506 mimics and the respective reporter plasmids were transfected into cells using Lipofectamine 3000 according to the manufacturer's protocol. After 24 hours, the Renilla and Fire y luciferase activities were determined using the Dual-Luciferase Reporter Assay System (Promega) according to the manufacturer's instructions.
RNA-binding protein immunoprecipitation (RIP) assay miR-506 mimics or miRNA was transfected into HCCC-9810 and CCLP1 cells. And the cells were lysed and collected in a RIP lysis buffer kit (Millipore, USA). Human anti-Ago2 (Millipore, USA) or mouse anti-IgG (Millipore, USA) were then conjugated with RNAs magnetic beads. Then the expression levels of puri ed RNA were determined by qRT-PCR.
The immunohistochemistry (IHC) assay IHC analysis was performed as previously described [16] . For this staining, after heat-induced epitope retrieval, para n-embedded sections were incubated with 3% H
RNA-binding protein immunoprecipitation (RIP) assay miR-506 mimics or miRNA was transfected into HCCC-9810 and CCLP1 cells. And the cells were lysed and collected in a RIP lysis buffer kit (Millipore, USA). Human anti-Ago2 (Millipore, USA) or mouse anti-IgG (Millipore, USA) were then conjugated with RNAs magnetic beads. Then the expression levels of puri ed RNA were determined by qRT-PCR.
The immunohistochemistry (IHC) assay IHC analysis was performed as previously described [16] . For this staining, after heat-induced epitope retrieval, para n-embedded sections were incubated with 3% H
About PubCompare
Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.
We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.
However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.
Ready to get started?
Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required
Revolutionizing how scientists
search and build protocols!