Overview

  • Product name

    Anti-Firefly Luciferase antibody
    See all Firefly Luciferase primary antibodies
  • Description

    Rabbit polyclonal to Firefly Luciferase
  • Host species

    Rabbit
  • Tested applications

    Suitable for: WB, ICC/IF, IP, Neutralisingmore details
  • Species reactivity

    Reacts with: Other species
  • Immunogen

    Recombinant full length protein corresponding to Firefly Luciferase aa 1-548. (Red Firefly (Luciola cruciata) Luciferase purified from E. coli).
    Sequence:

    MENMENDENIVVGPKPFYPIEEGSAGTQLRKYMERYAKLGAIAFTNAVTG VDYSYAEYLEKSCCLGKALQNYGLVVDGRIALCSENCEEFFIPVIAGLFI GVGVAPTNEIYTLRELVHSLGISKPTIVFSSKKGLDKVITVQKTVTTIKT IVILDSKVDYRGYQCLDTFIKRNTPPGFQASSFKTVEVDRKEQVALIMNS SGSTGLPKGVQLTHENTVTRFSHARDPIYGNQVSPGTAVLTVVPFHHGFG MFTTLGYLICGFRVVMLTKFDEETFLKTLQDYKCTSVILVPTLFAILNKS ELLNKYDLSNLVEIASGGAPLSKEVGEAVARRFNLPGVRQGYGLTETTSA IIITPEGDDKPGASGKVVPLFKAKVIDLDTKKSLGPNRRGEVCVKGPMLM KGYVNNPEATKELIDEEGWLHTGDIGYYDEEKHFFIVDRLKSLIKYKGYQ VPPAELESVLLQHPSIFDAGVAGVPDPVAGELPGAVVVLESGKNMTEKEV MDYVASQVSNAKRLRGGVRFVDEVPKGLTGKIDGRAIREILKKPVAKM


    Database link: P13129

  • Positive control

    • Red Firefly Luciferase transfected 293 whole cell lysate; Full length recombinant Red Firefly Luciferase; pCMV-Red Firefly Luciferase transfected 293 cells.

Properties

  • Form

    Liquid
  • Storage instructions

    Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C long term. Avoid freeze / thaw cycle.
  • Storage buffer

    Preservative: 0.05% Sodium azide
    Constituents: 69% PBS, 30% Glycerol, 0.1% BSA
  • Concentration information loading...
  • Purity

    Protein G purified
  • Clonality

    Polyclonal
  • Isotype

    IgG
  • Research areas

Applications

Our Abpromise guarantee covers the use of ab187340 in the following tested applications.

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

Application Abreviews Notes
WB 1/2000 - 1/4000. Predicted molecular weight: 60 kDa.
ICC/IF 1/500 - 1/1000.
IP Use at 25 µg/mg of lysate.
Neutralising Use at an assay dependent concentration.

Target

  • Relevance

    Luciferase from the firefly has become one of the more widely used reporter proteins for the study of gene expression. Luciferase catalyzes a bioluminescent reaction which requires the substrate luciferin as well as Mg2+ and ATP. Mixing these reagents with the cell extract containing luciferase, results in a flash of light that decays rapidly. This light can be detected by a luminometer. The total light emission is proportional to the luciferase activity of the sample.
  • Cellular localization

    Peroxisome
  • Alternative names

    • ec 1.13.12.7 antibody
    • Firefly antibody
    • Luciferase antibody
    • Luciferin 4 monooxygenase antibody

Images

  • Immunofluorescent analysis of Red Firefly Luciferase (from the Japanese firefly Luciola cruciata) in pCMV-Red Firefly Luciferase transfected (green, right panel) and untransfected 293 cells (left panel). Formalin-fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature. Cells were probed with ab187340 at 1/1000 dilution for at least 1 hour at room temperature, washed with PBS, and incubated with a DyLight 488-conjugated goat anti-rabbit IgG secondary antibody at 1/400 dilution for 30 minutes at room temperature. Nuclei (blue) were stained with Hoechst 33342 dye.

  • All lanes : Anti-Firefly Luciferase antibody (ab187340) at 1/4000 dilution

    Lane 1 : Non-transfected 293 whole cell lysate
    Lane 2 : Green Renilla Luciferase transfected 293 whole cell lysate
    Lane 3 : Red Firefly Luciferase transfected 293 whole cell lysate

    Blocking peptides at 10 µg per lane.

    Secondary
    All lanes : Goat anti-rabbit HRP at 1/500 dilution

    Developed using the ECL technique.

    Predicted band size: 60 kDa

  • All lanes : Anti-Firefly Luciferase antibody (ab187340) at 1/4000 dilution

    Lane 1 : Non-transfected 293 whole cell lysate
    Lane 2 : Red Firefly Luciferase transfected 293 whole cell lysate
    Lane 3 : North American firefly (Photynus pyralis) Luciferase transfected 293 whole cell lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    All lanes : Goat anti-rabbit HRP at 1/500 dilution

    Developed using the ECL technique.

    Predicted band size: 60 kDa

  • All lanes : Anti-Firefly Luciferase antibody (ab187340) at 1/4000 dilution

    Lane 1 : Full length recombinant Red Firefly Luciferase at 100 µg
    Lane 2 : Full length recombinant Red Firefly Luciferase at 50 µg
    Lane 3 : Full length recombinant Red Firefly Luciferase at 25 µg
    Lane 4 : Full length recombinant Red Firefly Luciferase at 10 µg

    Secondary
    All lanes : Goat anti-rabbit HRP at 1/500 dilution

    Developed using the ECL technique.

    Predicted band size: 60 kDa

  • Immunoprecipitation of Red Firefly Luciferase (from the Japanese firefly Luciola cruciata) was performed using whole cell lysates  from untransfected and Red Firefly or Green Renilla Luciferase transfected 293 cell lines. Antigen-antibody complexes were formed by incubating 200µg of lysate with 5µg of ab187340 overnight on a rocking platform at 4°C. The immune complexes were captured on 50ul Protein A/G Agarose, washed extensively, and eluted. Whole cell lysates from untransfected and transfected whole cell lysates (2µg) were used as controls. Samples were resolved on a 4-20% Tris-HCl polyacrylamide gel, transferred to a PVDF membrane, and blocked with 5% BSA/TBS-0.1%Tween-20 for 1 hour. The membrane was probed with ab187340 at 1/4000 dilution overnight rotating at 4°C.

     

    Lane 1: Non-transfected 293 whole cell lysate

    Lane 2: IP using ab187340 from non-transfected 293 whole cell lysate

    Lane 3: Green Renilla Luciferase transfected 293 whole cell lysate

    Lane 4: IP using ab187340 from Green Renilla Luciferase transfected 293 whole cell lysate

    Lane 5: Red Firefly Luciferase transfected 293 whole cell lysate
    Lane 6: IP using ab187340 from Red Firefly Luciferase transfected 293 whole cell lysate

  • Neutralizing effect of ab187340 on the activity of purified recombinant Red Firefly Luciferase (from the Japanese firefly Luciola cruciata). 10ng of recombinant enzyme were incubated with the indicated amounts of ab187340 for at least 30 min. Enzyme activity (RLU) was detected. Average RLU values from 10 timepoints per concentration of 4 replicates +/- standard deviation is shown.

References

ab187340 has not yet been referenced specifically in any publications.

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