Wenn Sie das Fenster schließen, wird Ihre Konfiguration nicht gespeichert, es sei denn, Sie haben Ihren Artikel in die Bestellung aufgenommen oder zu Ihren Favoriten hinzugefügt.
Klicken Sie auf OK, um das MILLIPLEX® MAP-Tool zu schließen oder auf Abbrechen, um zu Ihrer Auswahl zurückzukehren.
Wählen Sie konfigurierbare Panels & Premixed-Kits - ODER - Kits für die zelluläre Signaltransduktion & MAPmates™
Konfigurieren Sie Ihre MILLIPLEX® MAP-Kits und lassen sich den Preis anzeigen.
Konfigurierbare Panels & Premixed-Kits
Unser breites Angebot enthält Multiplex-Panels, für die Sie die Analyten auswählen können, die am besten für Ihre Anwendung geeignet sind. Unter einem separaten Register können Sie das Premixed-Cytokin-Format oder ein Singleplex-Kit wählen.
Kits für die zelluläre Signaltransduktion & MAPmates™
Wählen Sie gebrauchsfertige Kits zur Erforschung gesamter Signalwege oder Prozesse. Oder konfigurieren Sie Ihre eigenen Kits mit Singleplex MAPmates™.
Die folgenden MAPmates™ sollten nicht zusammen analysiert werden: -MAPmates™, die einen unterschiedlichen Assaypuffer erfordern. -Phosphospezifische und MAPmate™ Gesamtkombinationen wie Gesamt-GSK3β und Gesamt-GSK3β (Ser 9). -PanTyr und locusspezifische MAPmates™, z.B. Phospho-EGF-Rezeptor und Phospho-STAT1 (Tyr701). -Mehr als 1 Phospho-MAPmate™ für ein einziges Target (Akt, STAT3). -GAPDH und β-Tubulin können nicht mit Kits oder MAPmates™, die panTyr enthalten, analysiert werden.
.
Bestellnummer
Bestellinformationen
St./Pkg.
Liste
Dieser Artikel wurde zu Ihren Favoriten hinzugefügt.
Wählen Sie bitte Spezies, Panelart, Kit oder Probenart
Um Ihr MILLIPLEX® MAP-Kit zu konfigurieren, wählen Sie zunächst eine Spezies, eine Panelart und/oder ein Kit.
Custom Premix Selecting "Custom Premix" option means that all of the beads you have chosen will be premixed in manufacturing before the kit is sent to you.
Catalogue Number
Ordering Description
Qty/Pack
List
Dieser Artikel wurde zu Ihren Favoriten hinzugefügt.
Spezies
Panelart
Gewähltes Kit
Menge
Bestellnummer
Bestellinformationen
St./Pkg.
Listenpreis
96-Well Plate
Menge
Bestellnummer
Bestellinformationen
St./Pkg.
Listenpreis
Weitere Reagenzien hinzufügen (MAPmates erfordern die Verwendung eines Puffer- und Detektionskits)
Menge
Bestellnummer
Bestellinformationen
St./Pkg.
Listenpreis
48-602MAG
Buffer Detection Kit for Magnetic Beads
1 Kit
Platzsparende Option Kunden, die mehrere Kits kaufen, können ihre Multiplex-Assaykomponenten in Kunststoffbeuteln anstelle von Packungen erhalten, um eine kompaktere Lagerung zu ermöglichen.
Dieser Artikel wurde zu Ihren Favoriten hinzugefügt.
Das Produkt wurde in Ihre Bestellung aufgenommen
Sie können nun ein weiteres Kit konfigurieren, ein Premixed-Kit wählen, zur Kasse gehen oder das Bestell-Tool schließen.
Use Anti-NFκB Antibody, p65 subunit, active subunit, clone 12H11 (Mouse Monoclonal Antibody) validated in EMSA, FC, ICC, IF, IHC, IHC(P), WB to detect NFκB also known as Rel A.
More>>Use Anti-NFκB Antibody, p65 subunit, active subunit, clone 12H11 (Mouse Monoclonal Antibody) validated in EMSA, FC, ICC, IF, IHC, IHC(P), WB to detect NFκB also known as Rel A. Less<<
Anti-NFkB Antibody, p65 subunit Antibody, active subunit Antibody, clone 12H11
Alternate Names
Rel A
Background Information
The transcription factor NFkappaB (Nuclear Factor kappa B) is involved in the expression and regulation of a number of important cellular and physiological processes such as growth, development, apoptosis, immune and inflammatory response, and activation of various viral promoters including human immunodeficiency virus long terminal repeats. NFkappaB represents a group of structurally related and evolutionarily conserved proteins related to the proto-oncogene c-Rel with five members in mammals that include Rel (cRel), RelA (p65), RelB, NFkappaB1 (p50 and its precursor p105), and NFkappaB2 (p52 and its precursor p100). NFkappaB/Rel proteins exist as homo- or heterodimers to form transcriptionally competent or repressive complexes. Although most NFkappaB dimers are activators of transcription, the p50/50 and p52/52 homodimers can repress the transcription of their target genes. The p50/p65 heterodimer of NFkappaB is the most abundant in cells.
References
Product Information
Format
Purified
Control
TNF α-treated HeLa cells, PMA and calcium ionophore-treated Jurkat cells.
Presentation
Purified mouse monoclonal IgG3 liquid in buffer containing 0.02 M Phosphate buffer, 0.25 M NaCl, pH 7.6 with 0.1% sodium azide.
Use Anti-NFκB Antibody, p65 subunit, active subunit, clone 12H11 (Mouse Monoclonal Antibody) validated in EMSA, FC, ICC, IF, IHC, IHC(P), WB to detect NFκB also known as Rel A.
Key Applications
Electrophoretic Mobility Shift Assay
Flow Cytometry
Immunocytochemistry
Immunofluorescence
Immunohistochemistry
Immunohistochemistry (Paraffin)
Western Blotting
Application Notes
Immunofluorescence: A 1-10 μg/mL concentration of a previous lot was used in immunofluorescence.
Immunohistochemistry (paraffin sections): A 5-10 μg/mL (APAAP) concentration of a previous lot was used in immunohistochemistry.
Immunohistochemistry (frozen sections): A 5-10 μg/mL (APAAP) concentration of a previous lot was used in immunohistochemistry.
Immunohistochemistry: The clone 12H11 works best in fresh frozen or acetone fixed human tissues, however some groups have had reactivity in traditional formalin fixed tissue when the tissue is of human origin. It is recommended that ABC or enhanced detection systems be employed for the best visualization in either acetone or formalin fixed tissues. The antibody reacts with human tissues best. Rat tissue will also react but at a lower affinity, and the antibody does not react with mouse, other species have not been examined. Microwave citric acid buffer or trypsin digestion antigen recovery have both been successful with 12H11 on formalin fixed tissues.
Western blot: 5-10 µg/mL (ECL)
Electrophoretic Mobility Supershift Assay: A 0.5-1 μg/mL concentration of a previous lot was used in Supershift assay.
Flow cytometry: A previous lot of this antibody was used in flow cytometry. Fixed cells only, acetone fixed cells.
Optimal working dilutions must be determined by end user.
Biological Information
Immunogen
Peptide corresponding to human p65 coupled to BSA.
Epitope
p65 subunit, active subunit
Clone
12H11
Concentration
Please refer to lot specific datasheet.
Host
Mouse
Specificity
Recognizes an epitope overlapping the nuclear location signal (NLS) of the p65 subunit of the NFkB heterodimer. Thus it selectively binds to the activated form of NFkB (Zabel et al., 1993). The antibody can also be used for electrophoretic mobility supershift assays (EMSA). The nuclear factor kB (NFkB) is a sequence-specific DNA-binding protein (Sen & Baltimore, 1986). It is a pleiotropic transcription factor which is involved in the expression of a variety of cellular and viral genes (Lenardo & Baltimore, 1989). NFkB consists of two subunits which are named according to their molecular weight, p50 and p65 (Kawakami et al., 1988). The subunits are stabilized by a so-called inhibitory chain IkB (Baeuerle & Baltimore, 1988). In quiescent cells, NFkB resides in the cytosol in an inactive form which can be activated in vivo by treatment of cells with cytokines or protein kinase activators. In vitro, it is possible to generate the active form of NFkB by treatment with sodium deoxycholate, formadine or electrophoretic size fractionation. The active form of NFkB is a heterotetrameric protein, consisting of the two p50 and two p65 subunits (Lenardo et al., 1987). After activation NFkB translocates from the cytosol to the nucleus of the cell, binds to specific DNA sequences and initiates transcription.
Isotype
IgG3
Species Reactivity
Human
Mouse
Rat
Rabbit
Species Reactivity Note
Human, rat, and rabbit. Expected to react with mouse based on sequence homology.
The p50 (NFKB1)/p65 (RELA) heterodimer is the most abundant form of NFKB. The NFKB complex is inhibited by I-kappa-B proteins (NFKBIA, MIM 164008 or NFKBIB, MIM 604495), which inactivate NFKB by trapping it in the cytoplasm. Phosphorylation of serine residues on the I-kappa-B proteins by kinases (IKBKA, MIM 600664, or IKBKB, MIM 603258) marks them for destruction via the ubiquitination pathway, thereby allowing activation of the NFKB complex. Activated NFKB complex translocates into the nucleus and binds DNA at kappa-B-binding motifs such as 5-prime GGGRNNYYCC 3-prime or 5-prime HGGARNYYCC 3-prime (where H is A, C, or T; R is an A or G purine; and Y is a C or T pyrimidine).
Function: NF-kappa-B is a pleiotropic transcription factor which is present in almost all cell types and is involved in many biological processed such as inflammation, immunity, differentiation, cell growth, tumorigenesis and apoptosis. NF-kappa-B is a homo- or heterodimeric complex formed by the Rel-like domain-containing proteins RELA/p65, RELB, NFKB1/p105, NFKB1/p50, REL and NFKB2/p52 and the heterodimeric p65-p50 complex appears to be most abundant one. The dimers bind at kappa-B sites in the DNA of their target genes and the individual dimers have distinct preferences for different kappa-B sites that they can bind with distinguishable affinity and specificity. Different dimer combinations act as transcriptional activators or repressors, respectively. NF-kappa-B is controlled by various mechanisms of post-translational modification and subcellular compartmentalization as well as by interactions with other cofactors or corepressors. NF-kappa-B complexes are held in the cytoplasm in an inactive state complexed with members of the NF-kappa-B inhibitor (I-kappa-B) family. In a conventional activation pathway, I-kappa-B is phosphorylated by I-kappa-B kinases (IKKs) in response to different activators, subsequently degraded thus liberating the active NF-kappa-B complex which translocates to the nucleus. NF-kappa-B heterodimeric p65-p50 and p65-c-Rel complexes are transcriptional activators. The NF-kappa-B p65-p65 complex appears to be involved in invasin-mediated activation of IL-8 expression. The inhibitory effect of I-kappa-B upon NF-kappa-B the cytoplasm is exerted primarily through the interaction with p65. p65 shows a weak DNA-binding site which could contribute directly to DNA binding in the NF-kappa-B complex.
Subunit structure: Component of the NF-kappa-B p65-p50 complex. Component of the NF-kappa-B p65-c-Rel complex. Homodimer; component of the NF-kappa-B p65-p65 complex. Component of the NF-kappa-B p65-p52 complex. May interact with ETHE1. Binds AES and TLE1. Interacts with TP53BP2. Binds to and is phosphorylated by the activated form of either RPS6KA4 or RPS6KA5. Interacts with ING4 and this interaction may be indirect. Interacts with CARM1, USP48 and UNC5CL. Interacts with IRAK1BP1. Interacts with NFKBID. Interacts with NFKBIA. Interacts with GSK3B. Interacts with NFKBIB. Interacts with NFKBIE. Interacts with NFKBIZ. Part of a 70-90 kDa complex at least consisting of CHUK, IKBKB, NFKBIA, RELA, IKBKAP and MAP3K14. Interacts with HDAC3; HDAC3 mediates the deacetylation of RELA. Interacts with HDAC1; the interaction requires non-phosphorylated RELA. Interacts with CBP; the interaction requires phosphorylated RELA. Interacts (phosphorylated at 'Thr-254') with PIN1; the interaction inhibits p65 binding to NFKBIA. Interacts with SOCS1. Interacts with UXT. Interacts with MTDH and PHF11. Interacts with ARRB2. Interacts with human respiratory syncytial virus (HRSV) protein M2-1. Interacts with NFKBIA (when phosphorylated), the interaction is direct; phosphorylated NFKBIA is part of a SCF(BTRC)-like complex lacking CUL1.
Subcellular location: Nucleus. Cytoplasm. Note: Nuclear, but also found in the cytoplasm in an inactive form complexed to an inhibitor (I-kappa-B).
Post-translational modification: Ubiquitinated, leading to its proteosomal degradation. Degradation is required for termination of NF-kappa-B response. Phosphorylation on 'Ser-536' stimulates acetylation on 'Lys-310' and interaction with CBP; the phosphorylated and acetylated forms show enhanced transcriptional activity. Reversibly acetylated; the acetylation seems to be mediated by CBP, the deacetylation by HDAC3. Acetylation at 'Lys-122' enhances DNA binding and impairs association with NFKBIA. Acetylation at 'Lys-310' is required for full transcriptional activity in the absence of effects on DNA binding and NFKBIA association. Acetylation can also lower DNA-binding and results in nuclear export.
Routinely evaluated by Western Blot on PC12 lysates.
Western blot: 1:500 dilution of this lot detected NF KAPPA B, P65 on 10 μg of PC12 lysates.
Usage Statement
Unless otherwise stated in our catalog or other company documentation accompanying the product(s), our products are intended for research use only and are not to be used for any other purpose, which includes but is not limited to, unauthorized commercial uses, in vitro diagnostic uses, ex vivo or in vivo therapeutic uses or any type of consumption or application to humans or animals.
Storage and Shipping Information
Storage Conditions
Stable for 6 months at 2-8ºC from date of receipt. Aliquot to avoid repeated freezing and thawing. For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap.
