CDK1 Monoclonal Antibody for Western Blot, IF, IHC (F), IHC (P), Flow, IP and ELISA
CDK1 Monoclonal Antibody for Western Blot, IF, IHC (F), IHC (P), Flow, IP and ELISA
Immunohistochemistry analysis of CDC2/CDK1 showing staining in the cytoplasm and membrane of paraffin-embedded human breast carcinoma tissue (right) compared to a negative control without primary antibody (left). To expose target proteins, antigen retrieval was performed using 10 mM sodium citrate (pH 6.0), microwaved for 8-15 min. Following antigen retrieval, tissues were blocked in 3% H2O2-methanol for 15 min at room temperature, washed with ddH2O and PBS, and then probed with a CDC2/CDK1 mouse monoclonal Antibody (331800) diluted in 3% BSA-PBS at a dilution of 1:20 overnight at 4°C in a humidified chamber. Tissues were washed extensively in PBST and detection was performed using an HRP-conjugated secondary antibody followed by colorimetric detection using a DAB kit. Tissues were counterstained with hematoxylin and dehydrated with ethanol and xylene to prep for mounting.
Immunofluorescent analysis of CAP43 was performed using 70% confluent log phase HeLa cells treated with 50ng of TNF-alpha for 20 minutes. The cells were fixed with 4% paraformaldehyde for 10 minutes, permeabilized with 0.1% Triton™ X-100 for 10 minutes, and blocked with 1% BSA for 1 hour at room temperature. The cells were labeled with CAP43 Rabbit Polyclonal Antibody (42-6200) at 2ug/ml in 0.1% BSA and incubated for 3 hours at room temperature and then labeled with Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, Alexa Fluor® 488 conjugate (A27034) a dilution of 1:2000 for 45 minutes at room temperature (Panel a: green). Nuclei (Panel b: blue) were stained with SlowFade® Gold Antifade Mountant with DAPI (S36938). F-actin (Panel c: red) was stained with Alexa Fluor® 555 Rhodamine Phalloidin (Product # R415, 1:300). Panel d represents the merged image showing cytoplasmic localization. Panel e is untreated cell with less signal. Panel f represents control cells with no primary antibody to assess background. The images were captured at 60X magnification.