Nuclear Factor-KB P65 (NFkBP65) (N-Term), (NLS Domain) antibody

Details for Product No. ABIN117926
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Antigen
Synonyms c-Rel, C-Rel, Xrel2, c-rel, rel-A, v-rel, xrel, NFKB3, p65, NFkB, zgc:100833
Epitope
N-Term, NLS Domain
(66), (57), (46), (46), (36), (34), (33), (31), (30), (30), (28), (20), (19), (12), (11), (7), (7), (7), (7), (6), (6), (6), (4), (4), (4), (3), (3), (3), (3), (3), (3), (3), (3), (2), (2), (2), (2), (2), (2), (2), (2), (2), (2), (2), (2), (2), (1), (1), (1), (1), (1), (1), (1), (1), (1), (1), (1), (1), (1), (1), (1), (1), (1), (1), (1), (1), (1), (1), (1), (1), (1), (1), (1), (1), (1), (1), (1), (1), (1), (1), (1), (1), (1), (1), (1), (1), (1), (1), (1), (1), (1), (1), (1), (1), (1), (1), (1), (1), (1), (1), (1), (1), (1), (1)
Reactivity
Human
(854), (445), (354), (76), (75), (74), (38), (9), (8), (6), (2), (2), (1), (1)
Host
Rabbit
(799), (67), (6), (5), (1)
Clonality
Polyclonal
Conjugate
Un-conjugated
(24), (19), (15), (11), (10), (10), (10), (10), (10), (10), (10), (10), (10), (10), (1), (1), (1)
Application
ELISA, Electrophoretic Mobility-Shift Assay (EMSA), Immunocytochemistry (ICC), Immunofluorescence (IF), Western Blotting (WB)
(655), (336), (331), (170), (128), (123), (100), (37), (21), (18), (16), (12), (6), (5), (2), (1)
Pubmed 6 references available
Quantity 0.1 mg
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Catalog No. ABIN117926
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Immunogen This antibody was purified from whole rabbit serum prepared by repeated immunizations with the NFkappaB 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.
Sequence Peptide Sequence: R-I-E-E-K-R-K-R-T-Y-E-T
Specificity 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 NFkappaB heterodimer. Therefore, the antibody selectively binds to the activated form of NFkappaB. Anti-NFkappaB p65 NLS may react non-specifically with other proteins.
Purification Affinity Purified
Alternative Name RELA / NF-kB p65
Background NFkappaB 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 NFkappaB bound to IkappaB. NFkappaB 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 NFkappaB 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 IkappaB-alpha. IkappaB-alpha binds to the p65 subunit, preventing nuclear localization and DNA binding. IkappaB- alpha binding masks the NFkappaB nuclear localisation signal (NLS). A broad range of external stimuli lead to activation of NFkappaB and set off signalling cascades that ultimately converge on the IkappaB kinase (IKK) complex. Activated IKK specifically and directly phosphorylates IkappaB-alpha and this phophorylation event targets IkappaB-alpha for degradation. As a consequence, NFkappaB NLS is uncovered and nuclear translocation occurs.
Alternate names: NF kappa B p65, NFKB3, NFkB p65, Nuclear factor NF-kappa-B p65 subunit, Nuclear factor of kappa light polypeptide gene enhancer in B-cells 3, Rel A, Transcription factor p65
Gene ID 5970
NCBI Accession NP_001138610
UniProt Q04206
Application Notes Suitable for Immunohistochemistry, Immunoblotting, ELISA and supershift assays. Thisproduct was assayed by Immunoblot and found to be reactive against NFκB p65 NLS at adilution of 1: 2,000 followed by reaction with Peroxidase conjugated Affinity Purified
Restrictions For Research Use only
Format Liquid
Concentration 1.0 mg/mL (by UV absorbance at 280 nm)
Buffer 0.02 M Potassium Phosphate, 0.15 M Sodium Chloride, pH 7.2 with 0.01% Sodium Azide as preservative.
Preservative Sodium azide
Precaution of Use This product contains sodium azide: a POISONOUS AND HAZARDOUS SUBSTANCE which should be handled by trained staff only.
Handling Advice Dilute only prior to immediate use. Avoid cycles of freezing and thawing.
Storage -20 °C
Storage Comment Store vial at -20° C or below prior to opening. For extended storage aliquot contents and freeze at -20°C or below.
Expiry Date 12 months
Supplier Images
anti-Nuclear Factor-KB P65 (NFkBP65) (N-Term), (NLS Domain) antibody Immunofluorescencechemistry. Rabbit anti-p65 NLS was used at a 1:200 dilution to detect p65 in TNF stimulated DU145 cells. Image shown is at a 1:400 magnification. Tissue was fixed and prepared as for immunohistochemistry (see above).
anti-Nuclear Factor-KB P65 (NFkBP65) (N-Term), (NLS Domain) antibody (2) Immunohistochemistry. Rabbit anti-p65 NLS was used at a 1:200 dilution to detect p65 in (A) control DU145 cells and (B) TNF stimulated DU145 cells. Although DU145 show relatively high basal levels of nuclear p65 staining, significant enhancement of nuclear staining is seen in panel B as evidence of translocation and availability of the NLS to be bound by the antibody. Tissue was formalin-fixed, paraffin embedded followed by citrate retrieval. Blocking and hybridizationincluded 5%
Background publications Lenardo, Baltimore: "NF-kappa B: a pleiotropic mediator of inducible and tissue-specific gene control." in: Cell, Vol. 58, Issue 2, pp. 227-9, 1989 (PubMed).

Zabel, Henkel, Silva et al.: "Nuclear uptake control of NF-kappa B by MAD-3, an I kappa B protein present in the nucleus." in: The EMBO journal, Vol. 12, Issue 1, pp. 201-11, 1993 (PubMed).

Unlap, Jope: "Dexamethasone attenuates NF-kappa B DNA binding activity without inducing I kappa B levels in rat brain in vivo." in: Brain research. Molecular brain research, Vol. 45, Issue 1, pp. 83-9, 1997 (PubMed).

Delfino, Walker: "Hormonal regulation of the NF-kappaB signaling pathway." in: Molecular and cellular endocrinology, Vol. 157, Issue 1-2, pp. 1-9, 2000 (PubMed).

Zhang, Ghosh: "Molecular mechanisms of NF-kappaB activation induced by bacterial lipopolysaccharide through Toll-like receptors." in: Journal of endotoxin research, Vol. 6, Issue 6, pp. 453-7, 2001 (PubMed).

Lawlor, Alessi: "PKB/Akt: a key mediator of cell proliferation, survival and insulin responses?" in: Journal of cell science, Vol. 114, Issue Pt 16, pp. 2903-10, 2001 (PubMed).

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