Abcam
Goat Anti-Mouse IgG2a heavy chain (HRP)
ab97245
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I work with a protein of 26 kda which is close to the size of IgG light chain (25kDa). In immuno-precipitation, because the IP antibody will be in the final IP product, after de-naturing, the IgG heavy and light chain will be in the sample and eventually on the membrane. This means it will show up after blotting with a regular secondary antibody. If working with tumor lysates, blood is always in the sample. IgG, which is an abundant protein in blood, always exists in the sample. Thus I need a secondary antibody that binds to the heavy chain only, so that the band at ~26kDa is my protein interest, instead of the light chain of IgG. In my test result, Ab97245 reacts with mouse IgG2a heavy chain strongly. Ab97245 reacts with mouse IgG2a light chain slightly, which is the reason I did not give it a 5-star review. Ab97245 does not react with mouse IgG1 or human IgG, neither heavy chain nor light chain.
Western Blot
Mouse tumor lysates and mouse IgG1, mouse IgG2a, human IgG
No primary antibody used
5% milk in TBS buffer 1 hour at room temperature
1:5000 in TBST buffer 1 hour at room temperature
NA
ECL incubate at room temperature for 1 min
3 mouse tumor lysates and 3 different IgGs were loaded in gel. Tumor lysates have been also pre-cleared with protein A/G beads to get rid of most (but not all) IgG. No primary antibody was used. This secondary antibody was incubated 1:5000 at room temperature for 1 hour. Results Summary: 1. Ab97245 reacts with mouse IgG2a heavy chain strongly. 2. Ab97245 reacts with mouse IgG2a light chain slightly, which is the reason I did not give it a 5-star. 3. Ab97245 does not react with mouse IgG1 or human IgG, neither heavy chain or light chain.
Based on my experiences, IgG2a seems much more than IgG1 in mouse blood.
Reacts with mouse IgG 2a heavy chain strongly.
Still has a weak reaction with IgG2a light chain.
This secondary antibody will be very useful if you need your secondary to react with the heavy chain only.