Anti-Renilla Luciferase antibody

• ICC/IF

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Immunocytochemistry/ Immunofluorescence - Anti-Renilla Luciferase antibody (AB187338)

Immunofluorescent analysis of Renilla Luciferase (from the Sea pansy Renilla reniformis) in untransfected (left panel) and pCMV-Renilla Luciferase transfected (right panel) 293 cells. Formalin-fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature. Cells were probed with ab187338 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 1/400 for 30 minutes at room temperature. Nuclei (blue) were stained with Hoechst 33342 dye.

• FuncS (Neut/Block)

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Functional Studies (Neut/Block) - Anti-Renilla Luciferase antibody (AB187338)

Neutralizing effect of Renilla luciferase antibody on the activity of purified recombinant Renilla luciferase (from the Sea pansy Renilla reniformis) using a Renilla Luciferase Assay Kit. 1ng of recombinant enzyme were incubated with the indicated amounts of ab187338 for at least 30 min. Enzyme activity (RLU) was detected. Average RUI values from 10 timepoints per concentration of 4 replicates +/- standard deviation are shown.

• IP

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Immunoprecipitation - Anti-Renilla Luciferase antibody (AB187338)

Immunoprecipitation of Renilla Luciferase (from the Sea pansy Renilla reniformis) was performed using whole cell lysates from untransfected and Renilla Luciferase or Red Firefly luciferase transfected 293 cell lines. Antigen-antibody complexes were formed by incubating 200ug of lysate with 5ug of ab187338 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 (2ug) 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 ab187338 at a dilution of 1/4000 overnight rotating at 4°C.

Lane 1 : Non-transfected 293 whole cell lysate

Lane 2 : Renilla Luciferase IP from non-transfected 293 whole cell lysate

Lane 3 : Renilla Luciferase transfected 293 whole cell lysate

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

Lane 5 : Red Firefly Luciferase transfected 293 whole cell lysate

Lane 6 : Renilla Luciferase IP from Red Firefly Luciferase transfected 293 whole cell lysate

All lanes:

Immunoprecipitation - Anti-Renilla Luciferase antibody (ab187338)

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• WB

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Western blot - Anti-Renilla Luciferase antibody (AB187338)

All lanes:

Western blot - Anti-Renilla Luciferase antibody (ab187338) at 1/4000 dilution

Lane 1:

Full length recombinant Renilla Luciferase at 100 µg

Lane 2:

Full length recombinant Renilla Luciferase at 50 µg

Lane 3:

Full length recombinant Renilla Luciferase at 25 µg

Lane 4:

Full length recombinant Renilla Luciferase at 10 µg

Secondary

All lanes:

Goat anti-rabbit HRP at 1/500 dilution

Predicted band size: 36 kDa

true

• WB

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Western blot - Anti-Renilla Luciferase antibody (AB187338)

All lanes:

Western blot - Anti-Renilla Luciferase antibody (ab187338) at 1/4000 dilution

Lane 1:

Non-transfected 293 whole cell lysate at 10 µg

Lane 2:

Renilla Luciferase transfected 293 whole cell lysate at 10 µg

Lane 3:

Red Firefly Luciferase transfected 293 whole cell lysate at 10 µg

Secondary

All lanes:

Goat anti-rabbit HRP at 1/500 dilution

Predicted band size: 36 kDa

true

• WB

Supplier Data

Western blot - Anti-Renilla Luciferase antibody (AB187338)

All lanes:

Western blot - Anti-Renilla Luciferase antibody (ab187338) at 1/4000 dilution

Lane 1:

Non-transfected 293 whole cell lysate at 10 µg

Lane 2:

Renilla Luciferase transfected 293 whole cell lysate at 10 µg

Lane 3:

Not Green-shifted Renilla reniformis Luciferase transfected 293 whole cell lysate at 10 µg

Secondary

All lanes:

Goat anti-rabbit HRP at 1/500 dilution

Predicted band size: 36 kDa

true