Immunoprecipitation of an HA-tagged protein using Anti-HA Agarose (Product # 26182). HA-tagged GFP protein (from Pontellina plumata) was expressed with the Pierce Human In Vitro Protein Expression System and purified by incubation with 50µL of Anti-HA Agarose slurry in a Pierce Spin column (Product # 69705) along with 60µL of the in vitro translation reaction diluted to a final volume of 200µL in TBS for 1 hour at 4°C with end-over-end mixing. HA-tagged GFP was eluted from the resin by adding 3 x 1 column volume of 1mg/mL HA Peptide (Product # 26184). The elution fractions were combined and 20% of the elution fraction was electrophoresed in SDS-PAGE and stained with Gel Code Blue (Product # 24590). Immunoprecipitation eluate from human in vitro translation reaction containing no DNA used as a negative control (Lane 1); immunoprecipitation eluate from human in vitro translation reaction containing GFP-HA DNA (Lane 2); MW Marker (Lane 3).
|Tested species reactivity||Tag|
|Published species reactivity||Not Applicable|
|Host / Isotype||Mouse / IgG1|
|Immunogen||HA peptide YPYDVPDYA derivitized to ovalbumin|
|Storage buffer||PBS, pH 7.2|
|Contains||0.05% sodium azide|
|Tested Applications||Dilution *|
|Immunoprecipitation (IP)||20 - 100 µl|
* Suggested working dilutions are given as a guide only. It is recommended that the user titrate the product for use in their own experiment using appropriate negative and positive controls.
|Western Blot (WB)||See 1 publications below|
26182 is supplied as 10 ml of antibody on crosslinked 4% beaded agarose supplied as a 50% slurry in 0.1M phosphate, 0.15M NaCl, pH 7.2 with 0.05% sodium azide.
Specific for the HA peptide YPYDVPDYA. The anti-HA agarose has been used successfully in immunoprecipitation applications. The anti-HA agarose has 3.5mg mouse anti-HA IgG1 monoclonal antibody conjugated per mL of settled agarose resin.
HA Synthetic Peptide (Product # 26184) is available for use in neutralization and control experiments and to competitively elute HA-tagged fusion proteins from immobilized anti-HA affinity resin.
An HA tag is frequently incorporated into recombinant proteins for a variety of purposes. An anti-HA antibody can then be used to detect the protein when doing studies with transfected cells.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.
|Not Applicable||Not Cited||
Valproic acid causes proteasomal degradation of DICER and influences miRNA expression.
26182 was used in western blot to study the proteasomal degradation of DICER and altered miRNA levels induced by valproic acid
|Zhang Z,Convertini P,Shen M,Xu X,Lemoine F,de la Grange P,Andres DA,Stamm S||PloS one (8:null)||2013|