Time course showing induction of TNF-a signaling cascade upon treatment: Cellular localization of proteins in the NF-KB signaling pathway was detected upon treatment of HeLa cells with TNF-a (50 ng/ml) for 5, 10 and 25 min, respectively. Fixed and permeabilized cells were stained with ABfinity™ anti-IKK alpha/beta(pSpS176/180) Recombinant Rabbit Monoclonal Antibody (Product # 701643, 1 ug/ml) or anti-NF-kB Mouse Monoclonal Antibody (Product # 339900, 1 ug/ml) and labeled with Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, Alexa Fluor® 488 conjugate (Product # A27034, 0.4 ug/ml, 1:2500) and Goat anti-Mouse IgG (H+L) Superclonal™ Secondary Antibody, Alexa Fluor® 647 conjugate (Product # A28181, 0.4 ug/ml, 1:2500). Images show staining of phospho-IKK alpha/beta and NF-KB (panel a, e, i, m) in untreated cells. No significant basal levels of phosphorylated IKK alpha/beta (panel a; green) were detected. Treatment with TNF alpha led to an increase in the levels of phospho-IKK alpha/beta (panel b - d ; green) in the cytosol and the nucleus, and a corresponding translocation of NF-kB to the nucleus (panel f - h; red). The composite images are shown in panels i - l; green, red. Nuclei (blue) were stained using SlowFade® Gold Antifade Mountant with DAPI (Product # S36938, 1:50) and panel m - p; green, red represent the specific localization of the proteins with reference to DAPI. No background staining was observed in control cells with no prim
|Tested species reactivity||Rabbit|
|Host / Isotype||Goat / IgG|
|Immunogen||Recombinant full-length protein|
|Conjugate||Alexa Fluor® 488|
|Purification||Gravity column chromatography|
|Contains||5mM sodium azide|
|Storage Conditions||4° C, store in dark, DO NOT FREEZE!|
|Antibody Form||Whole Antibody|
|Tested Applications||Dilution *|
|Flow Cytometry (Flow)||1-5 µg/ml|
|Immunocytochemistry (ICC)||0.1-2 µg/ml|
|Immunofluorescence (IF)||0.1-2 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.
The sensitivity and specificity of each lot is confirmed using ELISA.
Minimal cross-reactivity with mouse, rat, human, bovine, guinea pig and donkey IgG is observed.
Superclonal™ secondary antibodies are developed to be highly specific for both heavy and light chains of IgG. Utilizing the power of recombinant technology and multiple phenotypic screens, Superclonal™ antibodies are selected for enhanced specificity to the target species IgG. These recombinant secondary antibodies show similar performance as traditional polyclonal secondary antibodies in common immunoassay applications, with minimal cross-reactivity to other commonly-used species. In addition, their recombinant animal-free-origin helps ensures lot-to-lot consistency, reducing the need to optimize each lot in relevant assays.
Superclonal™ secondary antibodies are designed to allow scientists to achieve the accurate results they need, without some of the concerns that can impact traditional polyclonal secondary antibodies.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.