Anti-rabbit IgG VeriBlot for IP secondary antibody (HRP) (ab131366)
- Product nameAnti-rabbit IgG VeriBlot for IP secondary antibody (HRP)See all IgG secondary antibodies ...
- SpecificityVeriBlot for IP secondary antibodies are anti-rabbit immunoblotting reagents that enable the trouble-free detection of immunoblotted target protein bands, without interference from denatured IgG. This allows to detect the (co-)immunoprecipitated protein without masking by the IgG heavy (50 kDa) and light chains (25 kDa). In general this interference tends to originate from secondary antibodies which recognize primary antibodies released with the antigen during the immunoprecipitation procedure or endogenous IgGs from the lysate itself. VeriBlot for IP secondary antibodies only recognize native (non-reduced) antibodies and therefore the detection of heavy and light chains is highly minimized, if the immunoprecipitate is fully reduced.
- Tested applicationsWB more details
- Storage instructionsStore at +4°C.
- Concentration information loading...
- Clonality Monoclonal
- General notesNumber of blots:
At least 20 (based on a 1:200 dilution in 5 ml milk).
Important protocol notes:
1. The anti-rabbit VeriBlot for IP secondary antibody (HRP) detects rabbit IgG polyclonal and monoclonal antibodies.
2. The anti-rabbit VeriBlot for IP secondary antibody (HRP) preferentially detects the non-reduced form of rabbit IgG over the reduced, SDS-deantured forms.
3. IP sample should be completely reduced/denatured before loaded onto a western blot.
4. Milk should be used as the blocking protein for the immunoblot.
Western blot and IP resources:
a) Western blot a beginner's guide
b) IP protocol
c) IP troubleshooting tips
- Research Areas
Our Abpromise guarantee covers the use of ab131366 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|WB||WB: 1/40 - 1/4000. The dilution will depend on the sensitivity of the HRP substrate. The dilution range recommended is 1:40 - 1:4000. Based on a 1:200 dilution (25 µL) in 5 ml milk researchers can perform 20 western blots. This product is recommended for the western blot detection of IP samples.|
Anti-rabbit IgG VeriBlot for IP secondary antibody (HRP) images
IP sample preparation: Histone H3 (mono methyl K9) was immunoprecipitated using 0.5mg Hela whole cell extract, 5µg of Rabbit polyclonal to Histone H3 (mono methyl K9) and 50µl of protein G magnetic beads (+). No antibody was added to the control (-). The antibody was incubated under agitation with Protein G beads for 10min, Hela whole cell extract lysate diluted in RIPA buffer was added to each sample and incubated for a further 10min under agitation. Proteins were eluted by addition of 40µl SDS loading buffer and incubated for 10min at 70oC;
Western blot conditions: 10µl of each sample was separated on a SDS PAGE gel, transferred to a nitrocellulose membrane, blocked with 5% BSA and probed with ab9045.
Secondary antibody: Anti-rabbit IgG VeriBlot for IP secondary antibody (HRP) (ab131366) at 1/1000 dilution.
References for Anti-rabbit IgG VeriBlot for IP secondary antibody (HRP) (ab131366)
ab131366 has not yet been referenced specifically in any publications.