Immunofluorescent analysis of NFkappaB/p65 (green) in HeLa cells either left untreated (left panel) or treated with 25ng/ml of TNF-alpha (right panel) for 25 minutes. Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature and blocked with 1% Blocker BSA (Product # 37525) for 15 minutes at room temperature. Cells were probed with a NFkappaB/p65 polyclonal antibody (Product # PA5-16545) at a dilution of 1:50 for at least 1 hour at room temperature, washed with PBS, and incubated with DyLight 488 goat anti-rabbit IgG secondary antibody (Product # 35552) at a dilution of 1:400 for 30 minutes at room temperature. F-Actin (red) was stained with DyLight 554 Phalloidin (Product # 21834) and nuclei (blue) were stained with Hoechst 33342 dye (Product # 62249). Images were taken on a Thermo Scientific ArrayScan or ToxInsight Instrument at 20X magnification.
|Tested species reactivity||Bovine, Dog, Fruit fly, Hamster, Human, Mouse, Pig, Rabbit, Rat, Xenopus|
|Published species reactivity||Rabbit, Rat, Human, Mouse, Not Applicable|
|Host / Isotype||Rabbit / IgG|
|Immunogen||A synthetic peptide from the C-terminus of human NFkappaB protein|
|Storage buffer||PBS, pH 7.4, with 0.2% BSA|
|Contains||0.09% sodium azide|
|Storage Conditions||4° C|
|Tested Applications||Dilution *|
|Immunofluorescence (IF)||1:50 - 1:200|
|Immunohistochemistry (Paraffin) (IHC (P))||1:50 - 1:100|
|Western Blot (WB)||5-10 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.
PA5-16545 targets NFkappaB/p65 in IHC (P), IF, and WB applications and shows reactivity with mouse, Rabbit, Rat, Xn, Bovine, Canine, Drosophila, Hamster, Human, and Porcine samples.
The PA5-16545 immunogen is a synthetic peptide from the C-terminus of human NFkappaB protein.
This antibody was orginally validated as part of a Thermo Scientific Cellomics High Content Screening Kit. The antibody sold separately may have slightly different performance and may need to be further optimized for the best results.
NFkappa B is a heterodimer consisting of a 50kDa DNA binding subunit and a 65kDa transactivating subunit (RelA). The p50/p65 heterodimer is present in the cytoplasm in inactive form bound to its inhibitor, IdeltaB. Upon stimulation of cells with stimuli like LPS, Interleukin-1, IkB gets phosphorylated and degraded by proteosomes releasing the activated NFkappaB which goes to the nucleus inducing gene expression. NFkappaB was originally identified as transcriptional activator for kappa light chain gene in B cells.
IP-MS enrichment of RELA (LFQ intensity): RELA was enriched 246-fold from A549 lysate compared to background proteins, using the optimized IP-MS workflow with Pierce MS-Compatible Magnetic IP Kit protein A/G (Part No. 90409) and RELA antibody (Part No. PA5-16545). The STRING database (www.string-db.org) was used to identify the protein interactor list. See more information on IP-MS verification of antibody selectivity. IP-MS validation info.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.
|Not Applicable||Not Cited||
Ectopic lymphoid structures function as microniches for tumor progenitor cells in hepatocellular carcinoma.
PA5-16545 was used in immunohistochemistry - paraffin section to study hepatocellular carcinoma and ectopic lymphoid structures function as microniches for tumor progenitor cells
|Finkin S,Yuan D,Stein I,Taniguchi K,Weber A,Unger K,Browning JL,Goossens N,Nakagawa S,Gunasekaran G,Schwartz ME,Kobayashi M,Kumada H,Berger M,Pappo O,Rajewsky K,Hoshida Y,Karin M,Heikenwalder M,Ben-Neriah Y,Pikarsky E||Nature immunology (16:1235)||2015|
Pulmonary cryptosporidiosis: role of COX2 and NF-kB.
PA5-16545 was used in immunohistochemistry - paraffin section to test if Cryptosporidium-induced changes in the lungs are mediated through the activation of COX-2 and NF-kB
|Asaad NY,Sadek GS||APMIS : acta pathologica, microbiologica, et immunologica Scandinavica (114:682)||2006|
Cardiac-Specific Activation of IKK2 Leads to Defects in Heart Development and Embryonic Lethality.
PA5-16545 was used in immunohistochemistry to characterize defects in heart development and embryonic lethality due to cardiac-specific activation of IKK2
|Kraut B,Maier HJ,Kókai E,Fiedler K,Boettger T,Illing A,Kostin S,Walther P,Braun T,Wirth T||PloS one (10:null)||2015|
Oral epithelial barrier function and the role of nuclear factor kappa-ß pathway in the pathogenesis of aphthous ulceration.
PA5-16545 was used in immunohistochemistry to study loss of oral epithelial barrier function in the pathogenesis of aphthous ulceration and the involvement of NFkB signaling pathways
|Günhan Ö,Günal A,Avc¿ A,Kurgan ¿,Dereci Ö,Kurt B,Celasun B||The Turkish journal of gastroenterology : the official journal of Turkish Society of Gastroenterology (24:508)||2014|
Nuclear transcription factor-kappa beta-dependent ultrastructural alterations within the placenta and systemic inflammatory activation in pregnant patients with hemolysis, elevated liver functions and low thrombocyte count (HELLP) syndrome: a case-control study.
PA5-16545 was used in immunohistochemistry to study the potential role of placental NFkB overexpression in the pathophysiology of HELLP syndrome
|Simsek Y,Gul M,Celik O,Aydin NE,Arda Düz S,Celik E,Ozerol E,Özerol IH,Tanbek K||Hypertension in pregnancy (32:281)||2013|
Surgical removal of endometrioma decreases the NF-kB1 (p50/105) and NF-kB p65 (Rel A) expression in the eutopic endometrium during the implantation window.
PA5-16545 was used in immunohistochemistry to study endometrial levels of NFkB1 and RelA following surgical endometrioma removal
|Celik O,Celik E,Turkcuoglu I,Yilmaz E,Ulas M,Simsek Y,Karaer A,Celik N,Aydin NE,Ozerol I,Unlu C||Reproductive sciences (Thousand Oaks, Calif.) (20:762)||2013|
Chemopreventive efficacy of green tea drinking against 1,2-dimethyl hydrazine-induced rat colon carcinogenesis.
PA5-16545 was used in immunohistochemistry to study the ability of drinking green tea to prevent against colon carcinogenesis in a rat model
|Sadik NA||Cell biochemistry and function (31:196)||2013|
Intestinal tumorigenesis initiated by dedifferentiation and acquisition of stem-cell-like properties.
PA5-16545 was used in immunohistochemistry to study the role of inflammatory signaling in the dedifferentiation and acquisition of stem-like properties associated with intestinal tumorigenesis
|Schwitalla S,Fingerle AA,Cammareri P,Nebelsiek T,Göktuna SI,Ziegler PK,Canli O,Heijmans J,Huels DJ,Moreaux G,Rupec RA,Gerhard M,Schmid R,Barker N,Clevers H,Lang R,Neumann J,Kirchner T,Taketo MM,van den Brink GR,Sansom OJ,Arkan MC,Greten FR||Cell (152:25)||2013|
Expression of nuclear factor kappa B and survivin in psoriasis.
PA5-16545 was used in immunohistochemistry to perform a comparative study of NFkB and survivin expression in normal and psoriatic epidermis
|Gunduz K,Temiz P,Gencoglan G,Inanir I,Catalkaya A||ISRN dermatology (2012:null)||2012|
Hepatic activation of IKK/NF¿B signaling induces liver fibrosis via macrophage-mediated chronic inflammation.
PA5-16545 was used in immunohistochemistry to study the role of chronic inflammation mediated by macrophages in the development of fibrosis following activation of hepatic IKK/NFkB
|Sunami Y,Leithäuser F,Gul S,Fiedler K,Güldiken N,Espenlaub S,Holzmann KH,Hipp N,Sindrilaru A,Luedde T,Baumann B,Wissel S,Kreppel F,Schneider M,Scharffetter-Kochanek K,Kochanek S,Strnad P,Wirth T||Hepatology (Baltimore, Md.) (56:1117)||2012|
Effect of Si-Miao-Yong-An on the stability of atherosclerotic plaque in a diet-induced rabbit model.
PA5-16545 was used in immunohistochemistry to study the ability of the Chinese herbal remedy Si-Miao-Yong-An to increase atherosclerotic plaque stability
|Peng L,Li M,Xu YZ,Zhang GY,Yang C,Zhou YN,Li LJ,Zhang JP||Journal of ethnopharmacology (143:241)||2012|
Hsp72 overexpression accelerates the recovery from caerulein-induced pancreatitis.
PA5-16545 was used in immunohistochemistry and western blot to study the ability of transgenic overexpression of HSP72 to speed recovery in a murine model of pancreatitis
|Lunova M,Zizer E,Kucukoglu O,Schwarz C,Dillmann WH,Wagner M,Strnad P||PloS one (7:null)||2012|
Potential role of p38-mitogene-activated protein kinase and nuclear factor-kappa B expression in testicular dysfunction associated with varicocele: an experimental study.
PA5-16545 was used in immunohistochemistry to study the roles of p38-MAPK and NFkB in a rat model of testicular dysfunction
|Simsek A,Ozbek E,Ilbey YO,Cekmen M,Somay A,Tasci AI||Andrologia (44 Suppl 1:94)||2012|
Extensive fibrin accumulation and accompanying epithelial changes in the pathogenesis of ligneous mucosal disease (ligneous periodontitis).
PA5-16545 was used in immunohistochemistry to study the pathogenesis of ligneous peridontitis and the role played by fibrin accumulation
|Günhan Ö,Avci A,Dereci Ö,Akgün S,Celasun B||The American Journal of dermatopathology (34:35)||2012|
Salmonella Engages Host MicroRNAs To Modulate SUMOylation: a New Arsenal for Intracellular Survival.
PA5-16545 was used in western blot to report that Salmonella enterica serovar Typhimurium modulates host SUMOylation.
|Verma S,Mohapatra G,Ahmad SM,Rana S,Jain S,Khalsa JK,Srikanth CV||Molecular and cellular biology (35:2932)||2015|
3,4-Dihydroxy-benzohydroxamic acid (Didox) suppresses pro-inflammatory profiles and oxidative stress in TLR4-activated RAW264.7 murine macrophages.
PA5-16545 was used in western blot to study the role of 3,4-Dihydroxy-benzohydroxamic acid (Didox) in acute inflammation and oxidative stress
|Matsebatlela TM,Anderson AL,Gallicchio VS,Elford H,Rice CD||Chemico-biological interactions (233:95)||2015|
Comparative gene expression profiling identifies common molecular signatures of NF-¿B activation in canine and human diffuse large B cell lymphoma (DLBCL).
PA5-16545 was used in western blot to perform a comparative study of the gene expression profiles of canine and human diffuse large B-cell lymphoma with particular reference to the NFkB pathway
|Mudaliar MA,Haggart RD,Miele G,Sellar G,Tan KA,Goodlad JR,Milne E,Vail DM,Kurzman I,Crowther D,Argyle DJ||PloS one (8:null)||2013|
Z-100, extracted from Mycobacterium tuberculosis strain Aoyama B, promotes TNF-¿ production via nucleotide-binding oligomerization domain containing 2 (Nod2)-dependent NF-¿B activation in RAW264.7 cells.
PA5-16545 was used in immunocytochemistry to evaluated the effects of Z-100 on NF-kB signaling in the murine macrophage cell line RAW264.7
|Katsunuma K,Yoshinaga K,Ohira Y,Eta R,Sato T,Horii T,Tanaka T,Takei M,Seto K||Molecular immunology (64:218)||2015|
|Not Applicable||Not Cited||
Identification of multipath genes differentially expressed in pathway-targeted microarrays in zebrafish infected and surviving spring viremia carp virus (SVCV) suggest preventive drug candidates.
PA5-16545 was used in immunocytochemistry to use zebrafish to identify factors altered during infection with spring viremia carp virus
|Encinas P,Garcia-Valtanen P,Chinchilla B,Gomez-Casado E,Estepa A,Coll J||PloS one (8:null)||2013|
Calpastatin upregulation in Mycoplasma hyorhinis-infected cells is promoted by the mycoplasma lipoproteins via the NF-¿B pathway.
PA5-16545 was used in immunocytochemistry and western blot to study the mechanism by which mycoplasma lipoproteins upregulate calpastatin in Mycoplasma hyorhinis-infected cells
|Elkind E,Vaisid T,Kornspan JD,Barnoy S,Rottem S,Kosower NS||Cellular microbiology (14:840)||2012|