BPTF/FALZ Antibody

A: All of the information I have shows that we successfully used this antibody in IP using this standard protocol, so it doesn't appear you need any special lysis buffer or treatment.

A: Regarding your question, nucleosome-remodeling factor subunit (BPTF) is a very large protein as you have also mentioned (338KD). There are some tips for efficient transfer of these proteins in western blot for instance: 1) Make sure to use a low percentage gel, for BPTF in particular I suggest you use 7% gel or even lower (5%). 2) You can use either Nitrocellulose or PVDF, to my knowledge the binding efficiency is higher for nitrocellulose while PVDF is a better choice of membrane if you are planning to do re-probing and stripping. If you are going to use PVDF, make sure to wet it in 100% methanol before use. Furthermore, PVDF membrane exhibits better binding efficiency of electroblotted material in the presence of SDS in the transfer buffer. However SDS added to facilitate transfer of large proteins should not exceed 0.05% as it will interfere with the binding of the protein to the membrane. 3) For transfer you can either use the semi-dry or wet transfer but you decide doing wet transfer, I strongly recommend you doing the transfer in low voltage (at 50v) for instance in order to obtain an efficient transfer. These are some suggestions I can provide for a better and more efficient transfer for large proteins like BPTF.

A: All of the information I have shows that we successfully used this antibody in IP using this standard protocol, so it doesn't appear you need any special lysis buffer or treatment.

A: Regarding your question, nucleosome-remodeling factor subunit (BPTF) is a very large protein as you have also mentioned (338KD). There are some tips for efficient transfer of these proteins in western blot for instance: 1) Make sure to use a low percentage gel, for BPTF in particular I suggest you use 7% gel or even lower (5%). 2) You can use either Nitrocellulose or PVDF, to my knowledge the binding efficiency is higher for nitrocellulose while PVDF is a better choice of membrane if you are planning to do re-probing and stripping. If you are going to use PVDF, make sure to wet it in 100% methanol before use. Furthermore, PVDF membrane exhibits better binding efficiency of electroblotted material in the presence of SDS in the transfer buffer. However SDS added to facilitate transfer of large proteins should not exceed 0.05% as it will interfere with the binding of the protein to the membrane. 3) For transfer you can either use the semi-dry or wet transfer but you decide doing wet transfer, I strongly recommend you doing the transfer in low voltage (at 50v) for instance in order to obtain an efficient transfer. These are some suggestions I can provide for a better and more efficient transfer for large proteins like BPTF.