Immunofluorescence analysis of TAK1 / MAP3K7 was done on 70% confluent log phase HeLa 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 TAK1 / MAP3K7 (28H25L68), ABfinity™ Rabbit Monoclonal Antibody (700113) 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 localization. Panel e is a no primary antibody control. The images were captured at 60X magnification.
|Tested species reactivity||Human|
|Host / Isotype||Rabbit / IgG|
|Immunogen||A recombinant protein corresponding to amino acids 476-606 of O43318.|
|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)||2-3 ug/ml|
|Immunofluorescence (IF)||2-3 ug/ml|
|Western Blot (WB)||0.5-2 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.
This antibody is predicted to react with Rhesus monkey, orangutan, chimpanzee, mouse, rat, equine, porcine, bovine, chicken, Xenopus and zebrafish 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.
TAK1 (also known as MAP3K7) is a versatile protein with many signaling functions. Originally identified as a TGF- beta-activated kinase, TAK1 interacts with TGF- beta-receptor and TRAF6 to modulate TGF- beta activation of JNK and p38. It has additional roles in activation of p38 and JNK through Wnt, BMP, and activin signaling pathways as well in response to thyroid hormone, osmotic stress, endothelin, and ephrine. Through activation of von Hippel-Lindau tumor suppressor expression, TAK1 represses PDGF-B, integrin beta1 and integrin beta5, promoting proper wound healing.TAK1 also plays an essential role in IKK activation in numerous signaling pathways including IL-1, IL-6, IL-18, TNF, CD40, TLR, and RIG-I. Activation of the TAK1-IKK pathway requires TAB2/3 and ubiquitination. In this process, TAB2/3 binds to the C-terminal region of TAK1 and becomes polyubiquitinated, TAK1 is autophosphorylated at Thr178, Thr184, Thr187 and Ser192, and finally TAK1 phosphorylates IKK- beta. Additionally, TAK1 represses human telomerase reverse transcriptase suggesting a role in regulation of cell lifespan.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.