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Immunofluorescence analysis of NFkB [p65] was done on 70% confluent log phase HeLa cells treated with TNF-alpha (50 ng/mL for 20 min). 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 NFkB [p65] Rabbit polyclonal Antibody (510500) at 2µg/mL in 1% BSA and incubated for 3 hours at room temperature and then labeled with Alexa Fluor 488 Goat Anti-Rabbit IgG Secondary Antibody (A11008) at a dilution of 1:400 for 30 minutes at room temperature (Panel a: green). Nuclei (Panel b: blue) were stained with SlowFade® Gold Antifade Mountant DAPI (S36938). F-actin (Panel c: red) was stained with Alexa Fluor 594 Phalloidin (A12381). Panel d is a merged image showing nuclear localization upon treatment. Panel e shows untreated HeLa cells with cytoplasmic localization. Panel f shows no primary antibody control. The images were captured at 20X magnification.
|Tested species reactivity||Human , Mouse , Primate|
|Published species reactivity||Rat , Human|
|Host / Isotype||Rabbit / IgG|
|Immunogen||A recombinant protein derived from the carboxyl terminal portion of the human NF-kappa-beta protein|
|Purification||Antigen affinity chromatography|
|Storage buffer||PBS, pH 7.4|
|Contains||0.1% sodium azide|
|Tested Applications||Dilution *|
|ChIP assay (ChIP)||5µg|
|ELISA (ELISA)||0.1-1.0 ug/ml|
|Flow Cytometry (Flow)||1:100-1:500|
|Immunohistochemistry (IHC)||0.5-2 ug/ml|
|Western Blot (WB)||1-3 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.
NF-kappaB (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-kappaB plays a key role in regulating the immune response to infection. Consistent with this role, incorrect regulation of NF-kappaB has been linked to cancer, inflammatory and autoimmune diseases, septic shock, viral infection and improper immune development. There are five members in the NF-kappaB family: NF-kappaB1, NF-kappaB2, RelA (also named p65), RelB, and c-Rel. The most common form of NF-kappaB is the p50/RelA heterodimer, although other forms of NF-kappaB dimers, such as p50/p50, p52/p52, p52/RelA, p50/c-Rel, c-Rel/c-Rel, p52/RelB, and p50/RelB, have also been identified in some types of cells. The primary role of NF-kappaB is to maintain normal cellular functions that range from cell-to-cell communication to cell motility, cell cycle progression, and cell lineage development. The activity of NF-kappaB is tightly regulated by interaction with inhibitory IkappaB proteins.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.
Expression of angiogenic factors vascular endothelial growth factor and interleukin-8/CXCL8 is highly responsive to ambient glutamine availability: role of nuclear factor-kappaB and activating protein-1.
||Bobrovnikova-Marjon EV,Marjon PL,Barbash O,Vander Jagt DL,Abcouwer SF||Cancer research (64:4858)||2004|
NF-kappa B in the vascular progression of melanoma.
||Kashani-Sabet M,Shaikh L,Miller JR,Nosrati M,Ferreira CM,Debs RJ,Sagebiel RW||Journal of clinical oncology : official journal of the American Society of Clinical Oncology (22:617)||2004|
PKC zeta participates in activation of inflammatory response induced by enteropathogenic E. coli.
||Savkovic SD,Koutsouris A,Hecht G||American journal of physiology. Cell physiology (285:C512)||2003|
Melatonin reduces renal interstitial inflammation and improves hypertension in spontaneously hypertensive rats.
||Nava M,Quiroz Y,Vaziri N,Rodriguez-Iturbe B||American journal of physiology. Renal physiology (284:F447)||2003|
NF-kappa-B p65delta3, nuclear factor NF-kappa-B p65 subunit, nuclear factor of kappa light polypeptide gene enhancer in B-cells 3, transcription factor p65, v-rel reticuloendotheliosis viral oncogene homolog A, avian reticuloendotheliosis viral (v-rel) oncogene homolog A, p65 NF kappaB, p65 NF-kappa B, p65 NFkB, RelA, NFKB3, Nuclear Factor kB p65 subunit, Transcription Factor p65