Immunofluorescence analysis of RIP2 was performed using 70% confluent log phase U-87 MG cells. The cells were fixed with 4% paraformaldehyde for 10 minutes, permeabilized with 0.1% Triton™ X-100 for 10 minutes, and blocked with 1% BSA for 1 hour at room temperature. The cells were labeled with RIP2 (AF28D3) Mouse Monoclonal Antibody (MA517221) at 2ug/ml in 0.1% BSA and incubated for 3 hours at room temperature and then labeled with Goat anti-Mouse IgG (H+L) Superclonal™ Secondary Antibody, Alexa Fluor® 488 conjugate (A28175) at a dilution of 1:2000 for 45 minutes at room temperature (Panel a: green). Nuclei (Panel b: blue) were stained with SlowFade® Gold Antifade Mountant with DAPI (S36938). F-actin (Panel c: red) was stained with Alexa Fluor® 555 Rhodamine Phalloidin (Product # R415, 1:300). Panel d represents the merged image showing cytoplasmic localization. Panel e shows the no primary antibody control. The images were captured at 60X magnification.
|Tested species reactivity||Human, Mouse, Rat|
|Host / Isotype||Mouse / IgG1|
|Immunogen||Recombinant human protein purified from E.coli (His-RIP2)|
|Storage buffer||HEPES with 0.15M NaCl, 0.01% BSA, 50% glycerol|
|Contains||0.03% sodium azide|
|Tested Applications||Dilution *|
|ELISA (ELISA)||Assay Dependent|
|Immunocytochemistry (ICC)||2 µg/ml|
|Immunofluorescence (IF)||2 µg/ml|
|Immunoprecipitation (IP)||2 ul|
|Western Blot (WB)||1 µg/ml|
* Suggested working dilutions are given as a guide only. It is recommended that the user titrate the product for use in their own experiment using appropriate negative and positive controls.
A suggested positive control for this product is HeLa, 293T, L929 or C6.
The RIP (receptor-interacting protein) family of serine/Threonine kinases (RIP-1,2,3,4,5,6,7) are crucial regulators of cell survival and cell death that can trigger pro-survival, inflammatory and immune responses via the activation of transcription factors (NF-kB and AP-1) and death-inducing programs. RIP2 (also known as RICK, CARDIAK, CCK and Ripk2) transduces signals from receptors of both immune responses. RIP2 carries a CARD at its C-terminal, which is essential for NF-kB activation. RIP2 is a critical downstream mediator of Nod1 and Nod2 signaling. Overexpression of RIP2 results in the activation of, in addition to NF-kB, the MAPKs JNK and ERK2, requiring its kinase activity to activate ERK2, but not JNK.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.