This antibody was verified by ELISA against peptide conjugated to BSA(MASMTGGQQMG/BSA).
Antibody concentration was determined by extinction coefficient: absorbance at280 nm of 1.4 equals 1.0 mg of IgG.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
Use at an assay dependent dilution. Use at a dilution of: Coating 1/100 - 1/500, Primary 1/1000 - 1/30,000.
Use at an assay dependent dilution.
1/1000 - 1/30000.
1/200 - 1/2000.
Use at an assay dependent dilution. ChIP was performed with 25 ug chromatin, 5ug of antibody and 20 ul of Protein A/G beads.
The T7 tag is an 11 amino acid peptide encoded in the leader sequence of T7 bacteriophage gene10. This gene encodes a T7 major capsid protein whose function is not clear. The T7 tag serves as a tag in many expression vectors including the pET system that is based on the very efficient T7 RNA polymerase expression system. Monoclonal antibodies specific for T7 tag are an important tool for studying expression of recombinant T7-tagged proteins.
A stably transfected 293T human cell line harbouring the GAL4 upstream activation sequence was transiently transfected with a V5 or T7- tagged GAL4 DNA Binding Domain construct. 48 hours post transfection Chromatin was prepared according to the Abcam X-ChIP protocol. The ChIP was performed with 25 ug chromatin, 5ug of antibody and 20 ul of Protein A/G beads. A non-specific antibody was used as the negative control. The immunoprecipitated DNA was quantified by real time PCR (SYBR Green approach).
Western blot - Anti-T7 tag® antibody - ChIP Grade (ab9138)
All lanes : Anti-T7 tag® antibody - ChIP Grade (ab9138) at 1/25000 dilution
Lane 1 : E. coli whole cell lysate expressing a multi-tag fusion protein at 0.2 µg Lane 2 : E. coli whole cell lysate expressing a multi-tag fusion protein at 0.1 µg Lane 3 : E. coli whole cell lysate expressing a multi-tag fusion protein at 0.05 µg