Immunofluorescence analysis of RELB was done on 70% confluent log phase HeLa cells. The cells were fixed with 4% paraformaldehyde for 15 minutes, permeabilized with 0.25% Triton™ X-100 for 10 minutes, and blocked with 5% BSA for 1 hour at room temperature. The cells were labeled with RELB Mouse Monoclonal Antibody (437500) at 1ug/mL in 1% BSA and incubated for 3 hours at room temperature and then labeled with Goat anti-Mouse Secondary Antibody, Alexa Fluor® 488 conjugate 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 Nuclear localization. Panel e is a no primary antibody control. The images were captured at 60X magnification.
|Tested species reactivity||Human, Mouse, Rhesus monkey|
|Host / Isotype||Mouse / IgG1, kappa|
|Immunogen||Recombinant protein derived from C-terminal region of human RelB protein.|
|Storage buffer||PBS, pH 7.4|
|Contains||0.1% sodium azide|
|Tested Applications||Dilution *|
|Immunocytochemistry (ICC)||1 µg/ml|
|Immunofluorescence (IF)||1 µg/ml|
|Immunohistochemistry (Paraffin) (IHC (P))||1:10-1:50|
|Immunoprecipitation (IP)||Assay Dependent|
|Western Blot (WB)||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.
NF-kB (Nuclear Factor kappa B) is a nuclear transcription factor found in all cell types and is involved in cellular responses to stimuli such as stress, cytokines, free radicals, ultraviolet irradiation, and bacterial or viral antigens. NF-kB plays a key role in regulating the immune response to infection. Consistent with this role, incorrect regulation of NF-kB has been linked to cancer, inflammatory and autoimmune diseases, septic shock, viral infection and improper immune development. There are five members in the NF-kB family: NF-kB1, NF-kB2, RelA (also named p65), RelB, and c-Rel. The RelB protein is present in the cytosol, bound to p50 or p52 and an inhibitory IkB protein, forming an inactive trimeric complex. Following cell signalling events leading to IkB degradation, Rel/NFk-B proteins are translocated to the nucleus where they regulate gene expression. The genes controlled by Rel/NF-kB family members are predominantly genes involved in the host response to infection, stress and injury. RelB mediates the regulation of genes involved in immune and inflammatory processes.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.