Anti-A. aeolicus BPL/BioID2 [SS 3A5-E2]

The SS 3A5-E2 monoclonal antibodies were generated to support the recent developments in the BioID, a method that can be used to detect potential interacting proteins

Contributors

Institute
A*STAR Accelerate Technologies Pte Ltd
Cat. #: 153485
Unit size: 100 ug
Research Fields: Tags and cell markers
Application: IF ; WB
Target: BPL R40G
Reactivity: Aquifex aeolicus
Clone: SS 3A5-E2
Host: Mouse
Class: Monoclonal
Alternate name: BPL antibody, BioID2 antibody, Anti-BPL, Anti-BioID
Product description: The SS 3A5-E2 monoclonal antibodies were generated to support the recent developments in the BioID, a method that can be used to detect potential interacting proteins . In the BioID method, a promiscuous biotin protein ligase (BPL) is fused to a protein of interests and expressed in vivo where it biotinylates proteins in a proximity-dependent manner. The biotinylated proteins can then be affinity purified and identified by mass spectrometry. The original BioID uses a promiscuous BPL from E.coli (BirA R118G), however due to its relatively large size it occasionally hindered proper targeting of the proteins it was fused to. The second generation of the BioID method is based on a BPL from hyperthermophilic bacterium Aquifex aeolicus (A. aeolicus) that was mutated within the conserved biotin binding site (R40G) causing loss of BPL single substrate specificity. The promiscuous A. aeolicus BPL R40G referred to as BioID2 is the smallest known BPL. The smaller BioID2 not only improved targeting of the bait but also proved to be more efficient in biotinylating proximate proteins.
Conjugation: Unconjugated
Isotype: IgG1 kappa
Molecular weight: 27 kDa
Immunogen: GST fused to A. aeolicus BPL R40G (BioID2)
Myeloma used: Sp2/0-Ag14
Target background: The SS 3A5-E2 monoclonal antibodies were generated to support the recent developments in the BioID, a method that can be used to detect potential interacting proteins . In the BioID method, a promiscuous biotin protein ligase (BPL) is fused to a protein of interests and expressed in vivo where it biotinylates proteins in a proximity-dependent manner. The biotinylated proteins can then be affinity purified and identified by mass spectrometry. The original BioID uses a promiscuous BPL from E.coli (BirA R118G), however due to its relatively large size it occasionally hindered proper targeting of the proteins it was fused to. The second generation of the BioID method is based on a BPL from hyperthermophilic bacterium Aquifex aeolicus (A. aeolicus) that was mutated within the conserved biotin binding site (R40G) causing loss of BPL single substrate specificity. The promiscuous A. aeolicus BPL R40G referred to as BioID2 is the smallest known BPL. The smaller BioID2 not only improved targeting of the bait but also proved to be more efficient in biotinylating proximate proteins.
Format: Liquid
Concentration: 1 mg/ml
Storage buffer: PBS with 0.02% azide
Storage conditions: -15° C to -25° C
Shipping conditions: Dry ice

Images

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Immunofluorescence staining of HeLa polyclonal cell line expressing BioID2 tagged to TorsinA ?E302/3 with SS 3A5-E2 monoclonal antibody. Cells were fixed in (A) cold methanol or (B) 4% PFA and stained with SS anti-BPL R40G/BioID2 3A5-E2 antibody (in green). The nuclei were counter-stained with Hoechst.

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