Anti-NFkB p50 may react non-specifically with other proteins. Control peptide (#R1008PEP) will compete only with the specific reaction of antiserum with Human NFkB p50 (NFKB1). In immunoblot no reaction was observed against the analogous Mmouse protein.
Purification
This product was prepared from monospecific antiserum by delipidation and defibrination.
Immunogen
Human NFkB p50 (NFKB1) peptide corresponding to a region near the N-terminus of the Human protein conjugated to Keyhole Limpet Hemocyanin (KLH)
Suitable for use in Immunoprecipitation, Immunoblotting (1/500-1/2,000), ELISA(1/5,000-1/25,000) and Supershift assays. This product was assayed by Immunoblot and found to be reactive against Human NFkBp50 (NFKB1) at a dilution of 1/1000 followed by reaction with Peroxidase conjugatedanti-Rabbit IgG [H&L] R1364HRP. Anti-Human NFkB p50 (NFKB1) is suitable for the detectionby Immunoblot of Human NFkB p50 (NFKB1) and its precursor protein p105. This product was also tested in a Gel Supershift assay and found to be reactive againstp50: p50 homodimers and p: 50: p65 heterodimers using 0.5 to 1.0 μL per assay. Gel (Super) Shift Information: In general, NFkB gel shift assays are assembled in 20 μL reactions containing 0.28 pmolesNFkB 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 ?g HeLa nuclear
Restrictions
For Research Use only
Format
Liquid
Concentration
80.0 mg/mL (by Refractometry)
Precaution of Use
This product contains sodium azide : a POISONOUS AND HAZARDOUS SUBSTANCE which should be handled by trained staff only.
Handling Advice
Avoid repeated freezing and thawing.
Storage
4 °C/-20 °C
Storage Comment
Store the antibody undiluted at 2-8 °C for one month or (in aliquots) at -20 °C for longer.
Target
NFKB1
(Nuclear Factor of kappa Light Polypeptide Gene Enhancer in B-Cells 1 (NFKB1))
NFκB 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. The heterodimers of p52/p65 and p50/p65 are regulated by physical inactivation in the cytoplasm by IkB-alpha. IkB-alpha binds to the p65 subunit, preventing nuclear localization and DNA binding. Low levels of p52 and p50 homodimers can also exist in cells.Synonyms: EBP-1, EBP1, KBF1, NF-kappa-B p50, NFKB1, Nuclear factor of kappa light polypeptide gene enhancer in B-cells 1