Anti-BioID2 antibody [SS 3A5-E2] (ab232733)

Overview

  • Product name

    Anti-BioID2 antibody [SS 3A5-E2]
  • Description

    Mouse monoclonal [SS 3A5-E2] to BioID2
  • Host species

    Mouse
  • Tested applications

    Suitable for: ICC/IF, WBmore details
  • Species reactivity

    Reacts with: Species independent
  • Immunogen

    Fusion protein corresponding to BioID2. (Glutathione-S-transferase fused to A. aeolicus BioID2 (BPL R40G)).

  • Positive control

    • ICC/IF: HeLa-BioID2-Dox cells. WB: BiolD2-transfected HeLa whole cell lysate
  • General notes

    BioID2 is a second generation biotin protein ligase (BPL) that can be used to identify protein-protein interactions. BioID2 is derived from the hyperthermophilic bacterium Aquifex aeolicus and was mutated within the conserved biotin binding site (R40G) causing promiscuous protein biotinylation. BioID2 is the smallest known BPL which improves targeting of the bait protein and improves labelling of other proteins in proximity (PubMed ID 26912792).

    This antibody clone is manufactured by Abcam. If you require a different buffer formulation or a particular conjugate for your experiments, please contact orders@abcam.com.

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: 0.02% Sodium azide
    Constituent: PBS
  • Concentration information loading...
  • Purity

    Protein G purified
  • Clonality

    Monoclonal
  • Clone number

    SS 3A5-E2
  • Isotype

    IgG1
  • Light chain type

    kappa

Applications

Our Abpromise guarantee covers the use of ab232733 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
ICC/IF Use a concentration of 1 µg/ml.

This antibody gives a signal in cells fixed with 4%PFA or 100% MeOH.

WB Use a concentration of 1 µg/ml.

Images

  • All lanes : Anti-BioID2 antibody [SS 3A5-E2] (ab232733) at 1 µg/ml

    Lane 1 : HeLa whole cell lysate
    Lane 2 : BiolD2-TorsinA transfected HeLa whole cell lysate, non-induced
    Lane 3 : BiolD2-TorsinA transfected HeLa whole cell lysate, DOX induced

    Lysates/proteins at 10 µg per lane.

    Performed under reducing conditions.

    Observed band size: 70 kDa
    why is the actual band size different from the predicted?



    This blot was produced using a 4-12% Bis-tris under the MOPS buffer system. The gel was run at 200V for 55 minutes before being transferred onto a Nitrocellulose membrane at 30V for 70 minutes. The membrane was blocked for an hour using 3% milk before ab232733 and ab181602 (Rabbit anti-GAPDH loading control) were incubated overnight at 4°C at a 1ug/ml concentration and 1/20000 dilution respectively. Antibody binding was detected using Goat anti-Mouse IgG H&L (IRDye® 800CW) preadsorbed (ab216772) and Goat anti-Rabbit IgG H&L (IRDye® 680RD) preadsorbed (ab216777) secondary antibodies at 1/20000 dilution for 1 hour at room temperature before imaging.

    Cell samples kindly provided by the Brian Burke laboratory, A-Star Institute, Singapore.

  • ab232733 staining BioID2 in HeLa cells expressing DOX-inducible BioID2 dystonia mutant of TorsinA. Parental HeLa cells have been used as negative control. The cells were fixed with 100% methanol (5min), permeabilized with 0.1% PBS-Triton X-100 for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1% PBS-Tween for 1h. The cells were then incubated overnight at +4°C with ab232733 at 1 μg/ml and ab6046, Rabbit polyclonal to beta Tubulin - Loading Control, at 1/1000 dilution. Cells were then incubated with ab150117, Goat Anti-Mouse IgG H&L (Alexa Fluor® 488) at 1/1000 dilution (shown in green) and ab150084, Goat polyclonal Secondary Antibody to Rabbit IgG - H&L (Alexa Fluor® 594) at 1/1000 dilution (shown in pseudocolor red). Nuclear DNA was labelled with DAPI (shown in blue).

    Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).

    Cell samples kindly provided by the Brian Burke laboratory, A-Star Institute, Singapore.

References

ab232733 has not yet been referenced specifically in any publications.

Customer reviews and Q&As

1-2 of 2 Abreviews or Q&A

Application
Western blot
Sample
Rat Cell lysate - whole cell (Neonatal rat ventricular cardiomyocytes)
Gel Running Conditions
Reduced Denaturing (10% polyacrylamide gel)
Loading amount
30 µg
Treatment
Transduced with adenovirus encoding for mycBioID2
Specification
Neonatal rat ventricular cardiomyocytes
Blocking step
Odyssey blocking buffer (LI-COR) in PBS as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 50% · Temperature: 25°C

Mr. Omer Shkedi

Verified customer

Submitted Feb 11 2019

Application
Immunocytochemistry/ Immunofluorescence
Sample
Rat Cell (Neonatal rat ventricular cardiomyocytes)
Permeabilization
Yes - 1% (v/v) Triton X-100 for 10 min.
Specification
Neonatal rat ventricular cardiomyocytes
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
Fixative
Formaldehyde

Mr. Omer Shkedi

Verified customer

Submitted Feb 11 2019

Please note: All products are "FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC PROCEDURES"
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