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MABS1341 Anti-N1-Phosphohistidine (1-pHis) Antibody, clone SC50-3

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      Key Specifications Table

      Species ReactivityKey ApplicationsHostFormatAntibody Type
      H, E. coliWB, DBRbPurifiedMonoclonal Antibody
      Catalogue NumberMABS1341
      DescriptionAnti-N1-Phosphohistidine (1-pHis) Antibody, clone SC50-3
      Alternate Names
      • N1-Phosphohistidine
      Background InformationPhosphorylation plays an important role in regulating protein activities and various cellular signaling events in cells. Limited by the tools available for phosphohistidine (pHis) detection, the majority of studies focus on serine, threonine, and tyrosine phosphorylations. Histidine phosphorylation can occur at either N1 (1-pHis) or N3 (3-pHis) of the imidazole ring. The development of peptides containing stable phosphoryltriazolylalanine analogues of 1-pHis and 3-pHis (1-pTza and 3-pTza) allows the generation of antibodies for studying both histidine N1 and N3 phosphorylations in signaling events. There is growing evidence implicating His kinases in cancer and tumor metastasis and the first metastasis suppressor gene identified is one of the two known mammalian His kinases, Nm23-H1 (also known as NME1, nucleoside diphosphate kinase, or NDPK-A). Nm23-H1/NME1 and the closely related Nm23-H2 (NME2/NDPK-B) catalyze the transfer of phosphate from ATP onto Nucleoside-diphosphates (NDPs) through a 1-pHis enzyme intermediate. Nm23-H1/-H2 also possess His kinase activity, transferring the phosphate from the active site pHis onto a His in a target protein. Metabolic enzymes such as phosphoglycerate mutase (PGAM), succinyl CoA synthase (SCS), and ATP citrate lyase (ACL) also use pHis as an enzyme intermediate. Unlike NME1/2, PGAM uses 3-pHis as an enzyme intermediate. In addition to eukaryotes, histidine phosphorylation is well documented in bacterial “two-component” signaling pathways involved in chemotaxis, although the phosphate is transferred from the pHis formed in the receptor/sensor protein to Asp residues of an acceptor response regulator protein, and the receptor/sensor protein essentially functions as an aspartate kinase.
      Product Information
      PresentationPurified rabbit monoclonal antibody in buffer containing 0.1 M Tris-Glycine (pH 7.4), 150 mM NaCl with 0.05% sodium azide.
      Quality LevelMQ100
      ApplicationAnti-N1-Phosphohistidine (1-pHis) antibody, clone SC50-3 is an isomer-specific monoclonal Ab to specifically detect histidine phosphorylated at position N1. This purified mAb is backed by published data demonstrating performance in Western blotting and dot blot applications.
      Key Applications
      • Western Blotting
      • Dot Blot
      Application NotesDot Blot Analysis: A representative lot detected peptides containing 1-pTza, but not 3-pTza phosphohistidine analogue. Clone SC50-3 displayed no immunoreactivity toward peptides containing only phosphorylated tyrosine or non-phosphorylated histidine (Fuhs, S.R., et al. (2015). Cell. 162(1):198-210).

      Western Blotting Analysis: Clone SC50-3 hybridoma culture supernatant was employed for Western blotting analysis of heat-sensitive histidine N1-phosphorylation (1-pHis) of exogenously expressed NM23-H1/NME1 fusion proteins in lysates from transformed E. coli and HEK293 transfectants (Fuhs, S.R., et al. (2015). Cell. 162(1):198-210).

      Note: DO NOT HEAT SAMPLES prior to phosphohistidine detection. Histidine phosphorylation is heat and acid labile. To generate negative control for specificity test, an aliquot of sample can be heated at 95ºC for 10-15 minutes to reverse histidine phosphorylation. Alternatively, an aliquot of sample can be incubated under acidified pH at 37ºC for 15 minutes to reduce histidine phosphorylation. Acidify each 100 µL sample with 25 µL of 1 M HCl before the incubation, then neutralize with 25 µL of 1 M NaOH prior to phosphohistidine detection.
      Biological Information
      ImmunogenKLH-conjugated library of random peptides containing non-hydrolyzable phosphohistidine analogue 1-pTza.
      EpitopeN1-phosphohistidine (1-pHis)
      ConcentrationPlease refer to lot specific datasheet.
      SpecificitySelectively detects proteins with histidine(s) phosphorylated at N1 of the imidazole ring (1-pHis), but not 3-pHis.
      Species Reactivity
      • Human
      • E. coli
      Species Reactivity NoteHuman, E. coli. Predicted to react with all species. Target modification is not species specific.
      Antibody TypeMonoclonal Antibody
      • Phosphorylation
      Purification MethodProtein A Purfied
      UniProt Number
      Molecular WeightVariable depending on the histidine-phosphorylated proteins.
      Physicochemical Information
      Materials Information
      Toxicological Information
      Safety Information according to GHS
      Safety Information
      Product Usage Statements
      Quality AssuranceEvaluated by Western Blotting of NME1 autophosphorylation reaction.

      Western Blotting Analysis: 0.24 µg/mL of this antibody detected recombinant human NME1 (NM23-H1) with N1-phosphohistidine (1-pHis) in a 5 µg aliquot of autophosphorylation reaction.
      Usage Statement
      • Unless otherwise stated in our catalog or other company documentation accompanying the product(s), our products are intended for research use only and are not to be used for any other purpose, which includes but is not limited to, unauthorized commercial uses, in vitro diagnostic uses, ex vivo or in vivo therapeutic uses or any type of consumption or application to humans or animals.
      Storage and Shipping Information
      Storage ConditionsStable for 1 year at 2-8°C from date of receipt.
      Packaging Information
      Material Size100 µL
      Transport Information
      Supplemental Information


      Anti-N1-Phosphohistidine (1-pHis) Antibody, clone SC50-3 SDS


      Safety Data Sheet (SDS) 

      Anti-N1-Phosphohistidine (1-pHis) Antibody, clone SC50-3 Certificates of Analysis

      TitleLot Number
      Anti-N1-Phosphohistidine (1-pHis), clone SC50-3 - 3211898 3211898
      Anti-N1-Phosphohistidine (1-pHis), clone SC50-3 - 3249539 3249539
      Anti-N1-Phosphohistidine (1-pHis), clone SC50-3 - 3322846 3322846
      Anti-N1-Phosphohistidine (1-pHis), clone SC50-3 -2659643 2659643
      Anti-N1-Phosphohistidine (1-pHis), clone SC50-3 -Q2574776 Q2574776
      Anti-N1-Phosphohistidine (1-pHis), clone SC50-3 Monoclonal Antibody 3090515
      Anti-N1-Phosphohistidine (1-pHis), clone SC50-3 Monoclonal Antibody 3012166
      Anti-N1-Phosphohistidine (1-pHis), clone SC50-3, Monoclonal Antibody 2770282
      Anti-N1-Phosphohistidine (1-pHis),clone SC50-3 -2848211 2848211


      Reference overviewPub Med ID
      Monoclonal 1- and 3-Phosphohistidine Antibodies: New Tools to Study Histidine Phosphorylation.
      Fuhs, SR; Meisenhelder, J; Aslanian, A; Ma, L; Zagorska, A; Stankova, M; Binnie, A; Al-Obeidi, F; Mauger, J; Lemke, G; Yates, JR; Hunter, T
      Cell  162  198-210  2015

      Show Abstract
      26140597 26140597

      Technical Info

      Characterization of Estrogen Receptor α Phosphorylation Sites in Breast Cancer Tissue Using the SNAP i.d® 2.0 System