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HMGN1 Antibody
SKU: orb345524
Description
Images & Validation
−Item 1 of 2
| Tested Applications | ELISA, IHC, WB |
|---|---|
| Dilution Range | ELISA: 1:5,000 - 1:25,000, IHC: 20 µg/ml, WB: 1:500 - 1:2,000 |
| Reactivity | Human |
| Application Notes |
Key Properties
−| Antibody Type | Primary Antibody |
|---|---|
| Host | Rabbit |
| Clonality | Polyclonal |
| Isotype | IgG |
| 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). |
| Target | HMGN1 |
| Purity | This affinity-purified antibody is directed against phosphorylated human HMGN protein at pS20 and pS24 residues (other HMGN proteins have this conserved sequence but at other residue positions). 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 cross-adsorbed 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. |
| Conjugation | Unconjugated |
Storage & Handling
−| Storage | Store vial at -20° C prior to opening. Aliquot contents and freeze at -20° C or below for extended storage. Avoid cycles of freezing and thawing. Centrifuge product if not completely clear after standing at room temperature. This product is stable for several weeks at 4° C as an undiluted liquid. Dilute only prior to immediate use. |
|---|---|
| Form/Appearance | Liquid (sterile filtered) |
| Buffer/Preservatives | Preservative: 0.01% (w/v) Sodium Azide. Stabilizer: None; Buffer: 0.02 M Potassium Phosphate, 0.15 M Sodium Chloride, pH 7.2 |
| Concentration | 1.20 mg/mL |
| Expiration Date | 12 months from date of receipt. |
| Dry Ice Shipping | Please note: This product requires shipment on dry ice. A dry ice surcharge will apply. |
| Disclaimer | For research use only |
Alternative Names
−rabbit anti-HMGN pS20/pS24 antibody, Non-histone chromosomal protein HMG-14, 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, HMGN-1, HMGN 1, HMG 14
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Biorbyt's affinity purified anti-HMGN pS20/pS24 antibody was used at 20 ug/ml to detect signal in a variety of tissues including multi-human, multi-brain and multi-cancer slides. This image shows moderate nuclear and faint cytoplasmic positive staining of epidermal keratinocytes at 40X. Tissue was formalin-fixed and paraffin embedded. The image shows localization of the antibody as the precipitated red signal, with a hematoxylin purple nuclear counterstain.
Western blot using Biorbyt's Affinity Purified anti-HMNG antibody shows detection of phosphorylated HMGN1 and HMGN2. Recombinant native and mutant HMGN proteins were treated with kinase PKCalpha to specifically phosphorylate S20 and S24 residues. Lanes contain: 1 - HMGN1, non-phosphorylated, 2 - HMGN1, phosphorylated, 3 - HMGN1 delta20E, delta24E, non-phosphorylated, 4 - HMGN1, delta20E, delta24E, phosphorylated, 5 - HMGN2, non-phosphorylated, and 6 - HMGN2, phosphorylated. Molecular weight markers (not shown) confirm the size of each recombinant protein. The primary antibody was diluted 1:1000 for this experiment. The blot was processed using a 5 sec exposure time.
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Protocol Information
WB
Western Blot (IB, immunoblot)
IHC
Immunohistochemistry
ELISA
Enzyme-linked Immunosorbent Assay (EIA)
HMGN1 Antibody (orb345524)
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