CREB Antibody (LB9)

Western Blot: CREB Antibody (LB9) [NB100-74393]

Western Blot: CREB Antibody (LB9) [NB100-74393] - Proteins were transferred to a PVDF membrane and blocked with 5% Milk/TBST for at least 1 hour. Membranes were probed with a mouse monoclonal antibody recognizing CREB at a dilution of 1:500 overnight at 4C on a rocking platform. Membranes were washed in TBS-0.1%Tween 20 and probed with a goat anti-mouse-HRP secondary antibody at a dilution of 1:20,000 for at least one hour. Membranes were washed and chemiluminescent detection performed.

Western Blot: CREB Antibody (LB9) [NB100-74393]

Western Blot: CREB Antibody (LB9) [NB100-74393] - Analysis of HepG2 cells

Immunohistochemistry-Paraffin: CREB Antibody (LB9) [NB100-74393]

Immunohistochemistry-Paraffin: CREB Antibody (LB9) [NB100-74393] - Cancer biopsies of deparaffinized human bladder and rectal tissue.

Chromatin Immunoprecipitation: CREB Antibody (LB9) [NB100-74393] - Analysis of CREB (green) in untreated HeLa cells. Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 15 minutes at room temperature. Cells were then blocked with 5% normal goat serum for 15 minutes at room temperature. Cells were probed with a mouse monoclonal antibody recognizing CREB at a dilution of 1:500 for at least 1 hour at room temperature. Cells were washed with PBS and incubated with DyLight 488 goat-anti-mouse secondary antibody at a dilution of 1:400 for 30 minutes at room temperature. Nuclei were stained with Hoechst 33342 dye. Images were taken on a Thermo Scientific ArrayScan at 20X magnification.

Immunoprecipitation: CREB Antibody (LB9) [NB100-74393]

Immunoprecipitation: CREB Antibody (LB9) [NB100-74393] - Analysis of CREB was performed on untreated 293T cells. Antigen: antbody complex was formed by binding 500ug whole cell lysate with 2ug of mouse monoclonal antibody recognizing CREB overnight on a rocking platform at 4C. The immune-complex was then captured on 50ul Protein A/G Plus Agarose. Captured immune-complexes were then washed extensively and proteins eluted with 5X Reducing Sample Loading Dye. Samples were then resolved on a 4-20% Tris-HCl polyacrylamide gel. Proteins were transferred to PVDF membrane and blocked with 5% Milk/TBS-0.1%Tween for at least 1 hour. Membranes were then probed with a mouse monoclonal antibody recognizing CREB at a dilution of 1:1000 overnight rotating at 4C. Membranes were then washed in TBST and probed with a goat anti-mouse-HRP secondary antibody at a dilution of 1:20,000 for at least one hour. Membranes were washed and chemiluminescent detection was performed using Super Signal West Dura.