NF-kB p65 Antikörper (N-Term)

Produktnummer ABIN105291

Hersteller Produkt- Nr.: 600-401-271

Produktdetails anti-NF-kB p65 Antikörper

Target: NF-kB p65 (NFkBP65) (Nuclear Factor-kB p65 (NFkBP65))

Bindungsspezifität: N-Term

Reaktivität: Human

Wirt: Kaninchen

Klonalität: Polyklonal

Konjugat: Dieser NF-kB p65 Antikörper ist unkonjugiert

Applikation: Western Blotting (WB), Immunohistochemistry (IHC), ELISA, Immunofluorescence (IF), Gel Shift (GS), Fluorescence Microscopy (FM)

Hersteller Produkt- Nr.: 600-401-271

Hersteller: Rockland

Verwendungszweck: NFkB p65 Antibody

Kreuzreaktivität (Details): This affinity-purified antibody is directed against the nuclear localization sequence (NLS) NLS of human p65 and is useful in determining its presence in various assays. The epitope recognized overlaps the NLS of the p65 subunit of the NFkB heterodimer.

Produktmerkmale: Synonyms: rabbit anti-NFKB p65 antibody, rabbit anti-p65 antibody, rabbit anti-NLS specific antibody, nuclear localization sequence, NFKB, NFkß, NF-kB, NF-kappaB, NFkappaB, Transcription factor p65, Nuclear factor NF-kappa-B p65 subunit, Nuclear factor of kappa light polypeptide gene enhancer in B-cells 3, RELA, NFKB3

Aufreinigung: Affinity purified antibody

Sterilität: Sterile filtered

Immunogen:

Immunogen: This antibody was purified from whole rabbit serum prepared by repeated immunizations with the NFkB p65 peptide corresponding to the NLS of the human protein conjugated to KLH using maleimide.  A residue of cysteine was added to the amino terminal end to facilitate coupling.

Immunogen Type: Conjugated Peptide

Isotyp: IgG

Anwendungsinformationen

Applikationshinweise:

Immunohistochemistry Dilution: 1:200

Gel Shift Dilution: 0.5 μL - 1.0 μL

Application Note: Anti-NFKB p65 antibody has been tested in ELISA, ICC, WB, and IF. NFkB gel shift assays are assembled in 20 μL reactions containing 0.28 pmoles NFkB oligo in 10 mM Tris ( pH 7.6), 50 mM NaCl, 0.5 mM EDTA, 1.0 mM DTT, 10 % glycerol. Some procedures specify the addition of 0.05 % NP-40. When using purified protein, 250-300 ng should be sufficient to produce a gel shifted complex, while 10 μg HeLa nuclear extract is utilized. The gel shift reactions are then incubated at room temperature for 30 minutes. The complexes are resolved on Tris-Glycine acrylamide gels. Loading dye containing bromophenol blue and xylene cyanol should be added to the negative control reaction only, as these dyes can increase the dissociation of the NFkB complexes. When using HeLa nuclear extract as the source of binding proteins, two sequence-specific gel-shifted complexes are expected, consisting of p50/p50 homodimers and p50/p65 heterodimers. For cells expressing p52, p50, and p65, as many as four sequence-specific gel-shifted complexes could be observed (p52/p52, p50/p50, p52/p65, p50/p65), and if high levels of p65 are present, the p65/p65 homodimer may also be weakly detected. The following reagents have been observed to enhance NFkB binding in vitro: millimolar amounts of GTP and ATP, spermine, spermidine, barium or calcium ions, and μM amounts of Co+3(NH3)6.

Western Blot Dilution: 1:2,000

ELISA Dilution: 1:5,000 - 1:25,000

IF Microscopy Dilution: 1:200

Other: User Optimized

Beschränkungen: Nur für Forschungszwecke einsetzbar

Handhabung

Format: Liquid

Konzentration: 1.0 mg/mL

Buffer:

Buffer: 0.02 M Potassium Phosphate, 0.15 M Sodium Chloride, pH 7.2

Stabilizer: None

Preservative: 0.01 % (w/v) Sodium Azide

Konservierungsmittel: Sodium azide

Vorsichtsmaßnahmen: This product contains Sodium azide: a POISONOUS AND HAZARDOUS SUBSTANCE which should be handled by trained staff only.

Lagerung: 4 °C,-20 °C

Informationen zur Lagerung: Store vial at -20° C prior to opening. Aliquot contents and freeze at -20° C or below for extended storage. Avoid cycles of freezing and thawing. Centrifuge product if not completely clear after standing at room temperature. This product is stable for several weeks at 4° C as an undiluted liquid. Dilute only prior to immediate use.

Haltbarkeit: 12 months

Referenzen für anti-NF-kB p65 Antikörper (ABIN105291)

Yu, Chang, Liu, Li, Kevork, Al-Hezaimi, Graves, Park, Wang: "Wnt4 signaling prevents skeletal aging and inflammation by inhibiting nuclear factor-κB." in: Nature medicine, Vol. 20, Issue 9, pp. 1009-17, (2014) (PubMed).

Graff, Ettayebi, Hardy: "Rotavirus NSP1 inhibits NFkappaB activation by inducing proteasome-dependent degradation of beta-TrCP: a novel mechanism of IFN antagonism." in: PLoS pathogens, Vol. 5, Issue 1, pp. e1000280, (2009) (PubMed).

Lord, Savitsky, Sitcheran, Calame, Wright, Ting, Williams: "Blimp-1/PRDM1 mediates transcriptional suppression of the NLR gene NLRP12/Monarch-1." in: Journal of immunology (Baltimore, Md. : 1950), Vol. 182, Issue 5, pp. 2948-58, (2009) (PubMed).

Liu, Ju, Willmarth, Casimiro, Ojeifo, Sakamaki, Katiyar, Jiao, Popov, Yu, Wu, Joyce, Wang, Pestell: "Nuclear factor-kappaB enhances ErbB2-induced mammary tumorigenesis and neoangiogenesis in vivo." in: The American journal of pathology, Vol. 174, Issue 5, pp. 1910-20, (2009) (PubMed).

Sivaramakrishnan, Niranjali Devaraj: "Morin regulates the expression of NF-kappaB-p65, COX-2 and matrix metalloproteinases in diethylnitrosamine induced rat hepatocellular carcinoma." in: Chemico-biological interactions, Vol. 180, Issue 3, pp. 353-9, (2009) (PubMed).

Liu, Li, Khoury, Colgan, Ibla: "Adenosine signaling mediates SUMO-1 modification of IkappaBalpha during hypoxia and reoxygenation." in: The Journal of biological chemistry, Vol. 284, Issue 20, pp. 13686-95, (2009) (PubMed).

Kumar, Wu, Collier-Hyams, Kwon, Hanson, Neish: "The bacterial fermentation product butyrate influences epithelial signaling via reactive oxygen species-mediated changes in cullin-1 neddylation." in: Journal of immunology (Baltimore, Md. : 1950), Vol. 182, Issue 1, pp. 538-46, (2008) (PubMed).

Abe, Hines, Zibari, Pavlick, Gray, Kitagawa, Grisham: "Mouse model of liver ischemia and reperfusion injury: method for studying reactive oxygen and nitrogen metabolites in vivo." in: Free radical biology & medicine, Vol. 46, Issue 1, pp. 1-7, (2008) (PubMed).

Khoury, Ibla, Neish, Colgan: "Antiinflammatory adaptation to hypoxia through adenosine-mediated cullin-1 deneddylation." in: The Journal of clinical investigation, Vol. 117, Issue 3, pp. 703-11, (2007) (PubMed).

Starkey, Haidacher, LeJeune, Zhang, Tieu, Choudhary, Brasier, Denner, Tilton: "Diabetes-induced activation of canonical and noncanonical nuclear factor-kappaB pathways in renal cortex." in: Diabetes, Vol. 55, Issue 5, pp. 1252-9, (2006) (PubMed).

Details zu NF-kB p65

Target: NF-kB p65 (NFkBP65) (Nuclear Factor-kB p65 (NFkBP65))

Andere Bezeichnung: RELA (NFkBP65 Produkte)

Hintergrund: Background: NFkB was originally identified as a factor that binds to the immunoglobulin kappa light chain enhancer in B cells.  It was subsequently found in non-B cells in an inactive cytoplasmic form consisting of NFkB bound to IkB. NFkB was originally identified as a heterodimeric DNA binding protein complex consisting of p65 (RelA) and p50 (NFKB1) subunits. Other identified subunits include p52 (NFKB2), c-Rel, and RelB. The p65, cRel, and RelB subunits are responsible for transactivation. The p50 and p52 subunits possess DNA binding activity but limited ability to transactivate. p52 has been reported to form transcriptionally active heterodimers with the NFkB subunit p65, similar to p50/p65 heterodimers. Low levels of p52 and p50 homodimers can also exist in cells. The heterodimers of p52/p65 and p50/p65 are regulated by physical inactivation in the cytoplasm by IkB-a. IkB-a binds to the p65 subunit, preventing nuclear localization and DNA binding.  IkB-a binding masks the NFkB nuclear localization signal (NLS).   A broad range of external stimuli lead to activation of NFkB and set off signaling cascades that ultimately converge on the IkB kinase (IKK) complex. Activated IKK specifically and directly phosphorylates IkB-a and this phosphorylation event targets IkB-a for degradation. As a consequence, NFkB NLS is uncovered and nuclear translocation occurs.

Gen-ID: 5970, 223468676

UniProt: Q04206

Pathways: NF-kappaB Signalweg, RTK Signalweg, T-Zell Rezeptor Signalweg, TLR Signalweg, Fc-epsilon Rezeptor Signalübertragung, Neurotrophin Signalübertragung, Activation of Innate immune Response, Cellular Response to Molecule of Bacterial Origin, Hepatitis C, Toll-Like Receptors Cascades, S100 Proteine