Immunofluorescence analysis of MDA5 was done on 70% confluent log phase A431 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 ABfinity™ MDA5(33H12L34), Recombinant Rabbit Monoclonal Antibody (700360) at 2ug/ml in 0.1% BSA and incubated for 3 hours at room temperature and then labeled with Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, Alexa Fluor® 488 conjugate (Product # A27034) 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 is a merged image showing cytoplasmic and nuclear localization. Panel e is a no primary antibody control. The images were captured at 60X magnification.
|Tested species reactivity||Human, Mouse|
|Published species reactivity||Human|
|Host / Isotype||Rabbit / IgG|
|Immunogen||A recombinant protein corresponding to amino acids 883-1025 of Q9BYX4.|
|Contains||0.09% sodium azide|
|Storage Conditions||Maintain refrigerated at 2-8°C for up to 1 month. For long term storage store at -20°C|
|Tested Applications||Dilution *|
|Flow Cytometry (Flow)||3-5 µg/10^6 cells|
|Immunocytochemistry (ICC)||4-6 ug/ml|
|Immunofluorescence (IF)||4-6 ug/ml|
|Immunohistochemistry (Paraffin) (IHC (P))||1:20|
|Western Blot (WB)||4-6 ug/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.
|Western Blot (WB)||See 1 publications below|
This antibody is predicted to react with Rhesus monkey, mouse, rat, porcine and bovine based on sequence homology.
ABfinity™ recombinant antibodies are rabbit monoclonal antibodies, unmatched for producing superior results. ABfinity™ antibodies are developed by immunizing animals, screening for functionality, cloning the immunogen-specific antibody genes into high-level mammalian expression vectors, produced on a large scale, and purified with Protein A.
ABfinity™ monoclonal antibodies resemble rabbit monoclonals isolated from serum or produced by hybridomas, but demonstrate greater specificity and sensitivity. Because ABfinity™ recombinant antibodies are derived from cloned DNA sequences of the heavy and light antibody chains, they are not susceptible to cell-line drift or lot-to-lot variation, thus allowing for peak specificity and performance.
Intact IgG appears on a non-reducing gel as ~150 kDa band and upon reduction generating a ~25 kDa light chain band and a ~50 kDa heavy chain.
Melanoma differentiation associated gene 5, MDA5, is a pattern recognition receptor to transmit the signal.Once activated, both proteins signal through IPS-1 activating transcription factors NF- kappaB and IRF-3 and ultimately activating apoptosis, cytokine signaling, and inflammation. RIG-I is essential for signaling by influenza A, influenza B, human respiratory syncytial virus, paromyxoviruses, Japanese encephalitis virus. MDA5 is essential for signaling by picornavirus. Both RIG-I and MDA5 are responsible for signaling by West Nile virus.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.
HepG2 cells mount an effective antiviral interferon-lambda based innate immune response to hepatitis C virus infection.
700360 was used in western blot to study the IFN-lambda inate immune response of HepG2 cells in response to HCV infection
|Israelow B,Narbus CM,Sourisseau M,Evans MJ||Hepatology (Baltimore, Md.) (60:1170)||2014|