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Anti-LgBiT Monoclonal Antibody

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Detects Large BiT (LgBiT) Subunit and LgBiT Fusion Proteins

  • Affinity-purified mouse monoclonal antibody
  • Use in Western blotting and immunofluorescence
  • Detects LgBiT subunit used in NanoBiT® complementation technology

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Catalog number selected: N7100

$ 398.00
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Anti-LgBiT Monoclonal Antibody
100μg
$ 398.00
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Detect LgBiT Subunit of NanoBiT® Complementation System

Anti-LgBiT Monoclonal Antibody is a protein A/G affinity-purified mouse monoclonal antibody that is used to detect Large BiT (LgBiT) and LgBiT fusion proteins via Western blotting and immunofluorescence. Weak cross-reactivity with NanoLuc® luciferase is observed.

LgBiT is the large 18kDa subunit of NanoBiT® complementation technology. LgBiT interacts with the low-affinity SmBiT or the high-affinity HiBiT peptides to form a functional luciferase. LgBiT protein fusions are used with SmBiT protein fusions for live cell analysis of protein interactions. Unfused LgBiT can also be expressed in cells to detect HiBiT-tagged proteins without the need for cell lysis.

Western Blot Results
The figure on the right shows an example Western blot using HEK293 cells transfected with CMV-based expression constructs encoding LgBiT fusion proteins. The expected mobility was seen for each of the LgBiT fusion proteins. We recommend a concentration of 1µg/ml as a starting point for protocol optimization. Low levels of the LgBiT-KRAS fusion were detected even with CMV-driven expression and without dilution using Transfection Carrier DNA.

Detection of NanoLuc Luciferase on Western Blot with Anti-NanoLuc Antibody

Detection of NanoLuc® fusion proteins expressed in HEK293 cells.

Immunofluorescence Results
Shown below are example immunofluorescent images of HeLa cells transfected with a CMV-based construct expressing LgBiT-CRAF fusion protein. The LgBiT-CRAF fusion protein is stained red, and nuclei are marked blue via Hoechst staining. We recommend a concentration of 1-3µg/ml for immunofluorescence optimization. No LgBiT staining is observed in HeLa cells transfected with only carrier DNA (mock).

LgBiT-CRAF 
(3µg/ml Anti-LgBiT)

LgBiT
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Hoescht
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Merge
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Mock (3µg/ml Anti-LgBiT)

LgBiT
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Hoescht
16616ta-screen-mock
Merge
16616ta-screen-mock

Specifications

What's in the box?

Item Part # Size

Anti-LgBiT Monoclonal Antibody

N710A 1 × 100μg

Certificate of Analysis

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Use Restrictions

For Research Use Only. Not for Use in Diagnostic Procedures.

Storage Conditions

BB

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