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Monoclonal Antibodies

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

Invented at A*STAR Accelerate Technologies Pte Ltd
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Image thumbnail for Anti-A. aeolicus BPL/BioID2 [SS 3A5-E2]
Image thumbnail for Anti-A. aeolicus BPL/BioID2 [SS 3A5-E2]
Image thumbnail for Anti-A. aeolicus BPL/BioID2 [SS 3A5-E2]
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Immunofluorescence staining of HeLa polyclonal cell line expressing BioID2 tagged to TorsinA ΔE302/3 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.
(A) Parental HEK293T or (B) monoclonal HEK293T cell line with BioID2 “knocked-in” into at least one LMNA gene alleles (resulting in BioID2 LaminA/C fusion protein) were immunofluorescently labelled with anti-BPL R40G/BioID2 SS 3A5-E2 monoclonal antibody (green). The nuclei were visualized with Hoechst.
Western blot analysis of SS 3A5-E2 monoclonal antibody. Total cell lysate of parental HeLa cell line (lane 1) or HeLa polyclonal cell line stably expressing BioID2-TorsinA ΔE302/3 without (lane 2) or with induced expression (lane 3) were transferred to a nitrocellulose membrane and blotted for BPL R40G/BioID2 with SS 3A5-E2 antibody. The predicted molecular weight of BioID2-TorsinA ΔE302/3 fusion protein is 65 kDa.
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Catalogue Number 153485
Applications IF WB
Antigen/Gene or Protein Targets BPL R40G
Synonyms BPL antibody, BioID2 antibody, Anti-BPL, Anti-BioID
Reactivity Aquifex aeolicus
Relevance 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.
Host Mouse
Immunogen GST fused to A. aeolicus BPL R40G (BioID2)
Positive Control Cells overexpressing A. aeolicus BPL R40G (BioID2) construct
Subclass IgG1 kappa
Molecular Weight (kDa) 27
Myeloma Used Sp2/0-Ag14
Strain C57BL/6
Notes This antibody is reactive with A. aeolicus and has not been tested with other species. The birA Entrez Gene number is NP_213397. UniProt number is O66837.
Research Area Epitope Tags

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Invented at

A*STAR Accelerate Technologies Pte Ltd
A*STAR Accelerate Technologies Pte Ltd

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