RIP abreview for Anti-Ago2 / eIF2C2 antibody - ChIP Grade

Good
Abreviews
Application
RIP
Sample
Mouse Cell lysate - whole cell (murine lung cancer cells)
Specification
murine lung cancer cells
Type
RIP without cross-linking
Duration of cross-linking step: 0 second(s)
Detection step
Real-time PCR
Positive control
empty vector cells for miR-10a
Negative control
RIP with control IgG
Immuno-precipitation step
Protein G

Other product details

Concentration
0.5 µg/10^6 cells
Incubation time
16 hour(s) and 0 minute(s) · Temperature: 4°C · Diluent: Millipore RIP Wash Buffer

Additional data

Additional Notes
The experiment involved the use of 368T1 murine lung cancer cells expressing either an empty vector (empty), a vector expressing wild-type Hmga2 (wt), a vector expressing a let-7 site mutated Hmga2 (m7), a vector expressing wild-type Hmga2 without a start codon (ATG wt), or a vector expressing a let-7 site mutated Hmga2 without a start codon (ATG m7).

I used the Millipore MAGNA-RIP kit with 5 ug Ago2 or IgG antibody per 1/2 of a 15 cm plate of cells (approximately 10^7 cells each). Following the RIPs, I did qRT-PCR using the Qiagen miScript system to detect miR-10a associated with IgG/Ago2 in these cells.

Overall, we observed no difference in miR-10a association between the samples but a clear enrichment (note the log scale) of miR-10a in the Ago2 IPs versus IgG.

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Submitted Nov 15 2012

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