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MABS1330 Anti-N1-Phosphohistidine (1-pHis) Antibody, clone SC1-1

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MABS1330
100 µL  
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      Overview

      Replacement Information

      Key Specifications Table

      Species ReactivityKey ApplicationsHostFormatAntibody Type
      H, E. coliWB, DB, ICC, IAPRbPurifiedMonoclonal Antibody
      Description
      Catalogue NumberMABS1330
      DescriptionAnti-N1-Phosphohistidine (1-pHis) Antibody, clone SC1-1
      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.
      References
      Product Information
      FormatPurified
      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
      Applications
      ApplicationAnti-N1-Phosphohistidine (1-pHis) antibody, clone SC1-1 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 Blots, immunofluorescence & immunoaffinity purification.
      Key Applications
      • Western Blotting
      • Dot Blot
      • Immunocytochemistry
      • Immunoaffinity Purification
      Application NotesDot Blot Analysis: A representative lot detected recombinant human NM23-H1/NME1 in vitro autophosphorylation on His118, as well as peptides containing 1-pTza, but not 3-pTza phosphohistidine analogue. Clone SC1-1 is most reactive toward 1-pTza peptides based on NM23-H1/NME1 1-pHis118 sequence, less reactive toward those based on histone H4 1-pHis18 or ACLY 1-pHis760 sequence, least reactive toward GNB1 1-pHis266-based or Kca3.1 1-pHis358-based sequence and not reactive toward phosphorylated tyrosine (Fuhs, S.R., et al. (2015). Cell. 162(1):198-210).
      Western Blotting Analysis: A representative lot detected heat-sensitive histidine N1-phosphorylation (1-pHis) in multiple cell lysates (Fuhs, S.R., et al. (2015). Cell. 162(1):198-210).
      Immunocytochemistry Analysis: A representative lot detected N1-phosphohistidine (1-pHis) immunoreactivity distinct from that of 3-pHis in 4% paraformaldehyde-fixed HeLa cells and murine bone marrow-derived macrophages by fluorescent immunocytochemistry. The 1-pHis immunoreactivity was found in regions surrounding acidic compartments, but not inside these compartments or nuclei (Fuhs, S.R., et al. (2015). Cell. 162(1):198-210).
      Immunoaffinity Purification: A representative lot was cross-linked to protein A resins for immunoaffinity purification of 1-pHis proteins from cell lysates prior to LC-MS/MS analysis (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)
      CloneSC1-1
      ConcentrationPlease refer to lot specific datasheet.
      HostRabbit
      SpecificitySelectively detects proteins with histidine(s) phosphorylated at N1 of the imidazole ring (1-pHis), but not 3-pHis.
      IsotypeIgG
      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
      Modifications
      • Phosphorylation
      Purification MethodProtein A Purfied
      UniProt Number
      Molecular WeightVariable depending on the histidine-phosphorylated proteins.
      Physicochemical Information
      Dimensions
      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: 0.3 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
      Specifications

      Documentation

      Anti-N1-Phosphohistidine (1-pHis) Antibody, clone SC1-1 SDS

      Title

      Safety Data Sheet (SDS) 

      Anti-N1-Phosphohistidine (1-pHis) Antibody, clone SC1-1 Certificates of Analysis

      TitleLot Number
      Anti-N1-Phos -Q2574775 Q2574775
      Anti-N1-Phosphohistidine (1-pHis), -2802034 2802034
      Anti-N1-Phosphohistidine (1-pHis), clone SC1-1 - 3059571 3059571
      Anti-N1-Phosphohistidine (1-pHis), clone SC1-1 - 3195958 3195958
      Anti-N1-Phosphohistidine (1-pHis), clone SC1-1 - 3258709 3258709
      Anti-N1-Phosphohistidine (1-pHis), clone SC1-1 - 3433915 3433915
      Anti-N1-Phosphohistidine (1-pHis), clone SC1-1 - 3524276 3524276
      Anti-N1-Phosphohistidine (1-pHis), clone SC1-1 - 3688982 3688982
      Anti-N1-Phosphohistidine (1-pHis), clone SC1-1 - 3799990 3799990
      Anti-N1-Phosphohistidine (1-pHis), clone SC1-1 -2687875 2687875

      References

      Reference overviewPub Med ID
      The protein histidine phosphatase LHPP is a tumour suppressor.
      Hindupur SK, Colombi M, Fuhs SR, Matter MS, Guri Y, Adam K, Cornu M, Piscuoglio S, Ng CKY, Betz C, Liko D, Quagliata L, Moes S, Jenoe P, Terracciano LM, Heim MH, Hunter T, Hall MN
      Nature  2018

      Show Abstract
      29562234 29562234
      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

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