Western Blot: BPTF/FALZ Antibody [NB100-41418] - Whole cell lysate (5, 15 and 50 mcg for WB; 1 mg for IP, 20% of IP loaded) from HeLa cells. NB100-41418 used for WB at 0.04 mcg/ml (A) and 0.1 mcg/ml (B) and used for IP ...read more
Immunohistochemistry-Paraffin: BPTF/FALZ Antibody [NB100-41418] - Human ovarian carcinoma. Antibody used at a dilution of 1:200.
Immunohistochemistry-Paraffin: BPTF/FALZ Antibody [NB100-41418] - FFPE section of human breast carcinoma. Affinity purified rabbit anti-FALZ/BPTF used at a dilution of 1:250.
The immunogen recognized by this antibody maps to a region between residue 1350 and 1400 of human Fetal Alzheimer Antigen (Bromodomain and PHD Domain Transcription Factor) using the numbering given in entry NP_872579.2 (GeneID 2186).
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Immunogen affinity purified
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Based on 100% sequence identity, this antibody is predicted to react with Orangutan, Chimpanzee and Northern white-cheeked gibbon.
Western Blot Image Notes: Detection of Human FALZ/BPTF by Western Blot and Immunoprecipitation. Samples:
Alternate Names for BPTF/FALZ Antibody
bromodomain and PHD domain transcription factor
Bromodomain and PHD finger-containing transcription factor
bromodomain PHD finger transcription factor
FAC1fetal Alz-50 reactive clone 1
FALZnucleosome-remodeling factor subunit BPTF
Fetal Alz-50 clone 1 protein
Fetal Alzheimer antigennucleosome remodeling factor, large subunit
FALZ/BPTF is a histone-binding component of NURF (nucleosome remodeling factor), a complex that catalyzes ATP-dependent nucleosome sliding and facilitates transcription of chromatin. This gene was identified in brain homogenates from patients with Alzheimer's disease. Analysis of the original protein (fetal Alz-50 reactive clone 1, or FAC1), containing a DNA-binding domain, a zinc finger motif, and a C-terminal bromodomain, suggested it might play a role in the regulation of transcription during proliferation. High levels of FAC1 were detected in fetal brain and in patients with neurodegenerative diseases. This protein is highly similar to the largest subunit of the Drosophila NURF (nucleosome remodeling factor) complex.
This product is for research use only and is not approved for use in humans or in clinical diagnosis. Primary Antibodies are guaranteed for 1 year from date of receipt.
FAQs for BPTF/FALZ Antibody (NB100-41418). (Showing 1 - 2 of 2 FAQ).
I am interesting by anti-BPTF antibody. I would like to know if a large protein like that could be revealed in PVDF membrane?
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.
Can you comment on BPTF being an histone bound protein, hence difficult to solubilize with a "simple" lysis buffer? You use here a NP-40 0.5% buffer also to be loaded directly for WB? Is the solubilization efficient enough?
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.