Rabbit HMGN Polyclonal Antibody | anti-HMGN antibody

Anti-HMGN Phospho-Ser20/Phospho-Ser24 Antibody

Cat. Num. AAA4157032

• Reactivity: Mouse, Rat, Bovine, Avian
• Applications: Western Blot, ELISA
• Purity: Affinity Purified
• Synonyms: HMGN; Polyclonal Antibody; Anti-HMGN Phospho-Ser20/Phospho-Ser24 Antibody; anti-HMGN antibody

• Host: Rabbit
• Reactivity: Mouse, Rat, Bovine, Avian
• Clonality: Polyclonal
• Isotype: IgG
• Specificity: This affinity-purified antibody is directed against the phosphorylated form of human HMGN1 protein at the pS20 and pS24 residues (other HMGN proteins have this conserved sequence although at other residue positions-see below). The product was affinity purified from monospecific antiserum by immunoaffinity purification. Antiserum was first purified against the phosphorylated form of the immunizing peptide. The resultant affinity purified antibody was then crossadsorbed against the non-phosphorylated form of the immunizing peptide. Reactivity occurs against human HMGN pS20/pS24 proteins and the antibody is specific for the phosphorylated form of the proteins. Reactivity is seen against HMGN1 and HMGN2. HMGN3 is also likely reactive although not tested. Reactivity with non-phosphorylated human HMGNs is minimal by ELISA. A BLAST analysis was used to suggest cross reactivity with HMGN proteins from Xenopus, chicken, mouse, bovine, dog, orangutan, rat and swine based on 100% homology with the immunizing sequence. Reactivity against homologues from other sources is not known.
• Purity/Purification: Affinity Purified
• Form/Format: Liquid; PBS, pH 7.4 with 0.05% sodium azide.
• Concentration: 1ug/ul (varies by lot)

• Applicable Applications for anti-HMGN antibody: Western Blot (WB), ELISA (EIA)
• Application Notes: This affinity purified antibody has been tested for use in ELISA and by western blot. Specific conditions for reactivity should be optimized by the end user. Expect a band approximately 17 kDa in size corresponding to phosphorylated HMGN proteins by western blotting in the appropriate cell lysate or extract. Less than 0.5% reactivity is observed against the non-phosphorylated form of the immunizing peptide. This antibody is phospho specific for pS20 and pS24 of HMGN proteins. The antibody was tested against HMGN1 by performing a standard phopshorylation in vitro assay using HMGN1 as a substrate and Msk1 as a kinase (known as a main effector kinase of MAPK signaling pathway and in vivo kinase for HMGN1). This kinase specifically phosphorylates S20 and S24 of HMGN1 (S24 and S28 of HMGN2). The reaction was resolved on SDS PAGE and immunoblotted, using antibody diluted at 1:1000 and 1:5,000. The antibody shows strong positive signal for Msk-phosphorylated HMGN1 and no cross-reaction for non-treated HMGN1.
• Immunogen: This affinity purified antibody was prepared from whole rabbit serum produced by repeated immunizations with a synthetic peptide corresponding to amino acids 19-28 of human HMGN protein.
• Preparation and Storage: This product is stable for several weeks at 4 degree C as an undiluted liquid. Dilute only prior to immediate use.; For extended storage, aliquot contents and freeze at-20 degree C or below. Avoid cycles of freezing and thawing. Expiration date is one (1) year from date of receipt.

Testing Data

Related Product Information for anti-HMGN antibody

HMGNs are proteins that bind chromatin effectively reducing the compaction of the chromatin fiber and enhancing access to DNA regulatory sequences. Members of this family have a conserved chromatin binding domain which is phosphorylated during mitosis. The sequence immunized is conserved in several species. As such, this reagent is designed as a-universal-reagent for the detection of all phosphorylated HMGN proteins. The High Mobility Group (HMG) proteins were originally isolated from mammalian cells and were named according to their electrophoretic mobility in polyacrylamide gels. HMGs were arbitrarily classed as a specific type of nonhistone proteins based on the observation that they are ubiquitous to mammalian cells, that they share certain physical properties, and that they are associated with isolated chromatin. HMG proteins and are now subdivided into 3 families: the HMGB (formerly HMG-1/-2) family, the HMGN (formerly HMG-14/-17) family, and the HMGA (formerly HMG-I/Y/C) family. Each HMG family has a characteristic functional sequence motif. The functional motif of the HMGB family is called the "HMG-box;" that of the HMGN family, the "nucleosomal binding domain;" and that of the HMGA family, the "AThook." The functional motifs characteristic of these canonical HMGs are widespread among nuclear proteins in a variety of organisms. Proteins containing any of these functional motifs embedded in their sequence are known as "HMG motif proteins."