Anti-mouse IgG VeriBlot for IP secondary antibody (HRP) (ab131368)
- Product nameAnti-mouse IgG VeriBlot for IP secondary antibody (HRP)See all IgG secondary antibodies ...
- Target speciesMouse
- SpecificityVeriBlot for IP secondary antibodies are anti-mouse 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 -20°C.
- Storage bufferConstituents: 1% Proprietary component, 99% Water
- Concentration information loading...
- Clonality Monoclonal
- General notesNumber of blots:
At least 20 blots (based on a 1 µl/ml dilution in 5 ml milk).
Important protocol notes:
1. The anti-mouse VeriBlot for IP secondary antibody (HRP) detects mouse IgG antibodies (subtypes: IgG1, IgG2a, IgG2b, IgG3).
2. The anti-mouse VeriBlot for IP secondary antibody (HRP) preferentially detects the non-reduced form of mouse IgG (IgG1, IgG2a, IgG2b, IgG3) over the reduced, SDS-denatured 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
- Research Areas
b) IP protocol
c) IP troubleshooting tips
Our Abpromise guarantee covers the use of ab131368 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|WB||WB: Use at an assay dependent concentration. The dilution will depend on the sensitivity of the HRP substrate. The recommended dilution is 1 µl in 1 ml milk. Based on the recommended dilution in 5 ml milk researchers can perform 20 western blots. This product is recommended for the western blot detection of IP samples.|
Anti-mouse IgG VeriBlot for IP secondary antibody (HRP) images
IP sample preparation: Caspase 7 was immunoprecipitated from 0.5 ml of 1x107 Jurkat cells/ml with 5 ug mouse anti-human Caspase 7.
WB conditions: Precipitate from 1x106 cells was subjected to electrophoresis, transferred to an PVDF membrane, and immunoblotted with an anti-Caspase 7 antibody.
Lane 1: Detection with anti-mouse IgG VeriBlot for IP secondary antibody (HRP) (ab131368)
Lane 2: Detection with an HRP-conjugated anti-mouse IgG H&L secondary antibody
Lane 3: Lane 1 was re-immunoblotted using an HRP-conjugated anti-mouse IgG H&L secondary antibody. The heavy and light-chains can now be seen, confirming that although the immunoprecipitating heavy and light-chains are present, ab131368 detects only native antibody and not denatured heavy and light-chains.
Please note the detection of the heavy and light-chains of the immunoprecipitating antibody in Lane 2 but not in Lane 1.
Developed using ECL technique under reducing conditions; exposure time 3 mins; blocking and antibody incubation steps done in 5% milk/TBST
2: Human heart homogenate Tissue Lysate – 20 µg
3: HeLa Cell Lysate – 20 µg
4: Mouse heart homogenate Tissue Lysate - 20 µg
5: NIH3T3 Cell Lysate – 20 µg
6: Rat heart homogenate Tissue Lysate – 20 µg
7: H9C2 cell lysate – 20 µg
Anti-Sodium ATPase antibody – Plasma Membrane Marker
Anti-GAPDH antibody – Cytosolic Marker
Anti-Histone H3 (di methyl k9) antibody – Nuclear Marker
Secondary: Anti-Mouse IgG VeriBlot for IP secondary antibody (ab131368) at 1/1000 dilution
Predicted Sodium Potassium ATPase band size: 112 kDa
Observed Sodium Potassium ATPase band size: 100 kDa
Predicted GAPDH band size: 37 kDa
Observed GAPDH band size: 37 kDa
Predicted Histone H3 (di methyl k9) band size: 17 kDa
Observed Histone H3 (di methyl k9 band size: 17 kDa
References for Anti-mouse IgG VeriBlot for IP secondary antibody (HRP) (ab131368)
ab131368 has not yet been referenced specifically in any publications.