Recombinant Human RNase H1 protein (ab153634)

Overview

Description

  • Nature
    Recombinant
  • Source
    Wheat germ
  • Amino Acid Sequence
    • Species
      Human
    • Sequence
      MSWLLFLAHRVALAALPCRRGSRGFGMFYAVRRGRKTGVFLTWNECRAQV DRFPAARFKKFATEDEAWAFVRKSASPEVSEGHENQHGQESEAKASKRLR EPLDGDGHESAEPYAKHMKPSVEPAPPVSRDTFSYMGDFVVVYTDGCCSS NGRRRPRAGIGVYWGPGHPLNVGIRLPGRQTNQRAEIHAACKAIEQAKTQ NINKLVLYTDSMFTINGITNWVQGWKKNGWKTSAGKEVINKEDFVALERL TQGMDIQWMHVPGHSGFIGNEEADRLAREGAKQSED
    • Amino acids
      1 to 286
    • Tags
      proprietary tag N-Terminus

Specifications

Our Abpromise guarantee covers the use of ab153634 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

  • Applications

    ELISA

    Western blot

  • Form
    Liquid
  • Additional notes

    Protein concentration is above or equal to 0.03 mg/ml.

  • Concentration information loading...

Preparation and Storage

  • Stability and Storage

    Shipped on dry ice. Upon delivery aliquot and store at -80ºC. Avoid freeze / thaw cycles.

    pH: 8.00
    Constituents: 0.31% Glutathione, 0.79% Tris HCl

General Info

  • Alternative names
    • H1RNA
    • MGC108918
    • Ribonuclease H type II
    • RibonucleaseH1
    • RNaseH1
    • RNH1
    see all
  • Relevance
    Rnase H1 specifically degrades the RNA of RNA-DNA hybrids.
  • Cellular localization
    Cytoplasmic

Images

  • ab153634 on a 12.5% SDS-PAGE stained with Coomassie Blue.

References

ab153634 has not yet been referenced specifically in any publications.

Customer reviews and Q&As

Application
Functional Studies
We tested enzymatic activity of Human RNase H1 full length protein (ab153634). RNase H1 is an endonuclease that cleaves RNA in DNA-RNA duplex. To see the protein's cleavage activity, we used 363 nt 5'-32P labeled RNA that contains target sequence of our antisense oligonucleotide. We observed expected sized RNA fragments after 5 min incubation of the RNA-oligonucleotide duplex with the RNase H1 protein at 37°C (Figure A). We also compare the reaction with E.coli RNase H. The result shows that the human RNase H1 gives the same sized fragment as E. coli RNase H does (Figure B).
Method:
RNase H mediated cleavage of 5’-32P-RNA (363 nt). (A) 5’-32P-RNA (0.5 pmol) was mixed with 20 nt antisense oligonucleotides (3 pmol) in buffer containing 60 mM Tris-HCl (pH7.8), 60 mM KCl, 2.5 mM MgCl2, and 2 mM DTT. The mixtures were heated at 95 °C for 3 min and slowly cooled to room temperature. Subsequently, various amount of human RNase H1 was added to the mixtures and incubated at 37 °C for 5 min. the reactions were quenched by adding an equal amount of loading dye containing deionized formamide. The samples were heated at 95 °C for 5 min, then quickly cooled down on ice and loaded onto 5% polyacrylamide gel containing 7 M urea. (B) 5’-32P-RNA (0.5 pmol) was mixed with 20 nt antisense oligonucleotides (3 pmol) in buffer containing 60 mM Tris-HCl (pH7.8), 60 mM KCl, 2.5 mM MgCl2, and 2 mM DTT. The mixtures were heated at 95 °C for 3 min and slowly cooled to room temperature. Subsequently, 2.5 units E. coli RNase H was added to the mixtures and incubated at 37 °C. the reactions were quenched at 2, 5, 10, 20 and 30 min by adding an equal amount of loading dye containing deionized formamide. The samples were heated at 95 °C for 5 min, then quickly cooled down on ice and loaded onto 5% polyacrylamide gel containing 7 M urea.

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Submitted Dec 29 2015

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