Anti-Halo Tag, derived from DhaA of Rhodococcus rhodochrous

Product no: AS21 4605

AS21 4605  | Clonality:  Polyclonal |  Host:  Rabbit  | Reactivity: Recombinant proteins which include Halo Tag®

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  • Product Info
  • Immunogen: KLH-conjugated peptide derived from DhaA of Rhodococcus rhodochrous, so called HaloTag®.
    Host: Rabbit


    Purity: Antigen affinity purified serum, in PBS pH 7.4


    Quantity: 50 µg
    Reconstitution: For reconstitution add 50 µl of sterile or deionized water.
    Storage: Store lyophilized/reconstituted at -20°C; once reconstituted make aliquots to avoid repeated freeze-thaw cycles. Please, remember to spin tubes briefly prior to opening them to avoid any losses that might occur from lyophilized material adhering to the cap or sides of the tubes.
    Tested applications: Western blot (WB)
    Recommended dilution: 1 : 1000 (WB)
  • Reactivity
  • Confirmed reactivity: Recombinant proteins with HaloTag®
    Predicted reactivity: Recombinant proteins with HaloTag®
  • Application Examples
  • Western blot detection of a recombinant protein using anti-Halo epitope tag antibodies

    1. Ladder (PageRuler Prestained, 10 à 180 kDa - Thermo)
    2. Non induced bacteria
    3. Induced bacteria at 37°C for 3h
    4. Aliquot lysate (after sonication) supernatant
    5. Flowthrough after incubation with beads
    6. Elution 1 with acidic elution buffer
    7. Elution 2 with acidic elution buffer
    8. Elution of beads with 2x SDS-sample buffer

    Recombinant protein was purified from 25ml E. coli Rossetta bacteria induced at 37°C during 3h. After sonication of the culture (lysate), Halo-Trap Agarose beads (chromotek) were added to the supernatant and incubated during 1h at 4°C. After 3 wash, beads were eluted with an acidic buffer (x2) and with 50 μl laemmli 2X). Samples were then separated on 12 % SDS-PAGE and blotted for 1 h to nitrocellulose (pore size of 0,45 um), using wet transfer. Blot was blocked with 5 % milk for 2h/RT with agitation. Blot was incubated in the primary antibody at a dilution of 1: 1000 in 5 % milk TBS-T ON/4°C with agitation. The antibody solution was decanted, and the blot was washed 4 times for 5 min in TBS-T at RT with agitation. Blot was incubated with secondary antibody (AS09 602, Rabbit anti-mouse IgG, HRP conjugated) diluted to 1: 20000 in 5 % milk for 1h/RT with agitation. The blot was washed as above and developed with a following chemiluminescent detection reagent AS16 ECL-S-10, AgriseraBright. Exposure time was 30 seconds.

    Courtesy of Paul Schouvelier, IBPS, Equipe Biologie de Semences, France

  • Additional Information
  • Additional information: HaloTag® is a trademark of Promega Corporation.
  • Background
  • Background: HaloTag is derived from the haloalkane dehalogenase enzyme DhaA of Rhodococcus rhodochrous and can be incorporated to a protein of interest (POI) and facilitate cellular and biochemical analysis.. HaloTag® is a trademark of Promega Corporation.
  • Protocols
  • Agrisera Western Blot protocol and video tutorials

    Protocols to work with plant and algal protein extracts

    Agrisera Educational Poster Collection
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AS09 602 |  Clonality: Polyclonal | Host: Goat | Reactivity: Rabbit IgG (H&L)

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AS21 4696 | Clonality: Monoclonal |  Host: Mouse | Reactivity: Green Fluorescence Protein (GFP)

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Buy 2 items of this product for 239.00 €/items
Buy 3 items of this product for 217.00 €/items