In Simple Western only 10-15 uL of the recommended dilution is used per data point.
The observed molecular weight of the protein may vary from the listed predicted molecular weight due to post translational modifications, post translation cleavages, relative charges, and other experimental factors.
Complement component C3 plays a central role in the activation of complement system. Its activation is required for both classical and alternative complement activation pathways. People with C3 deficiency are susceptable to bacteria infection.
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FAQs for Complement C3 (4)
I am trying to establish a method to measure mice C3 levels by nephelometry. I would be most grateful if you could provide me with some help, regarding the choice of the Ab. I totally understand that since such a method has never been tried, I do not expect any guaranties.
We have never performed nephelometry in our lab, and do not have a protocol or advice to provide about this application. However, it seems to me that you should choose an antibody that is capable of recognizing its target in its folded conformation. Therefore, I would suggest trying an antibody that has been validated for ICC or IHC.
At what molecular weight can we find the band when run on western blotting with and with out reducing agent?
The theoretical molecular weight of Complement C3 is 187kDa. With NBP1-32080, we typically see bands around 180kDa when testing in Western blot under reducing conditions. As we have not tested this antibody in non-reducing conditions we cannot say with certainty at what size you will see the band, but it will most likely be at a higher weight than the band in reducing conditions.
In the immunohistochemistry paraffin section protocol....the PBS buffer....should it be at certain pH?
For IHC application, different labs cites the pH of PBS buffer in the range of 7.1 - 7.6 but most commonly used pH is pH 7.4 (this is what we use in our lab). You may use this PBS (pH 7.4) for making permeablization buffer, antibody diluent buffers and for making wash buffer. For more on the protocol that we use in our lab and for IHC-P troubleshooting suggestions, you may visit: IHC-P Protocol and IHC-P Troubleshooting.
I was wondering what does Format 7C mean as far as anitbodies go?
7C is a fluorophore - it is useful for FLOW. The details of the fluor are (A=425, E=500)