Western Blotting (WB), Immunohistochemistry (IHC), ELISA
Characteristics
Concentration Definition: by UV absorbance at 280 nm
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 (see below).
This affinity purified antibody has been tested for use in ELISA, immunohistochemistry 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 1:1,000 and 1:5,000. The antibody shows strong positive signal for Msk-phosphorylated HMGN1 and no cross-reaction for non-treated HMGN1.
Restrictions
For Research Use only
Format
Liquid
Concentration
1.20 mg/mL
Buffer
0.02 M Potassium Phosphate, 0.15 M Sodium Chloride, pH 7.2
Preservative
Sodium azide
Precaution of Use
This product contains sodium azide: a POISONOUS AND HAZARDOUS SUBSTANCE which should be handled by trained staff only.
Storage
-20 °C
Target
HMGN1
(High Mobility Group Nucleosome Binding Domain 1 (HMGN1))
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Background
This antibody is designed, produced, and is suitable for Cancer, Immunology and Nuclear Signaling research. 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 ''AT-hook.'' 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.'' Synonyms: High mobility group nucleosome binding domain containing protein 1 antibody, High mobility group protein 14 antibody, High-mobility group nucleosome binding domain 1 antibody, HMG14 antibody