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
- Tested applicationsWB more details
- Storage instructionsStore at +4°C.
- Storage bufferpH: 7.2
Constituents: 0.16% Sodium phosphate, 0.88% Sodium chloride, 50% Glycerol, 0.1% BSA
- 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.