Human/Mouse/Rat Casein Kinase 1 alpha Antibody

Detection of Human and Rat Casein Kinase 1 alpha by Western Blot. Western blot shows lysates of HeLa human cervical epithelial carcinoma cell line, K562 human chronic myelogenous leukemia cell line, NRK rat normal kidney cell line, and Y3-Ag rat myeloid cell line. PVDF membrane was probed with 1 µg/mL of Human/Mouse/Rat Casein Kinase 1a Antigen Affinity-purified Polyclonal Antibody (Catalog # AF4569) followed by HRP-conjugated Anti-Sheep IgG Secondary Antibody (HAF016). A specific band was detected for Casein Kinase 1a at approximately 38 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.

Western Blot Shows Human Casein Kinase 1 alpha Specificity Using Knockdown Cell Line. Western blot shows lysates of HCT 116 human colorectal carcinoma cell line and Casein Kinase 1 alpha knockdown HCT 116 cell line (KD). Nitrocellulose membrane was probed with 2 µg/mL of Human/Mouse/Rat Casein Kinase 1 alpha Antigen Affinity-purified Polyclonal Antibody (Catalog # AF4569) followed by HRP-conjugated donkey anti-sheep IgG Secondary Antibody. A specific band was detected for Casein Kinase 1 alpha at approximately 36 kDa (as indicated) in the parental HCT 116 cell line, but is significantly reduced in knockdown HCT 116 cell line. The Ponceau stained transfer of the blot is shown. This experiment was conducted under reducing conditions. Image, protocol, and testing courtesy of YCharOS Inc. See ycharos.com for additional details.

Detection of Casein Kinase 1 alpha by Immunoprecipitation. HCT 116 lysates were prepared and immunoprecipitation was performed using 2.0 µg of Human/Mouse/Rat Casein Kinase 1 alpha Antigen Affinity-purified Polyclonal Antibody (Catalog # AF4569) pre-coupled to Dynabeads protein G. Immunoprecipitated Casein Kinase 1 alpha was detected with a Rabbit Anti-Casein Kinase 1 alpha antibody. The Ponceau stained transfer of the blot is shown. SM=4% starting material; UB=4% unbound fraction; IP=immunoprecipitate; HC=antibody heavy chain. Image, protocol and testing courtesy of YCharOS Inc. (ycharos.com).

Casein Kinase 1 alpha Specificity is Shown by Immunocytochemistry in Knockdown Cell Line. HCT 116 WT and Casein Kinase 1 alpha KD cells were labelled with a green or a far-red fluorescent dye, respectively. Cells were stained with Human/Mouse/Rat Casein Kinase 1 alpha Antigen Affinity-purified Polyclonal Antibody (Catalog # AF4569) followed by incubation with a donkey anti-sheep Alexa-fluor 555 coupled secondary antibody (upper panel). DAPI-only counterstained cells shown on a lower panel. Acquisition of the blue (nucleus-DAPI), green (identification of WT cells), red (antibody staining) and far-red (identification of KD cells) channels was performed. Representative images of the blue and red (grayscale) channels are shown. WT and KD cells are outlined with green and magenta dashed line, respectively. Primary antibody concentration used: 2 µg/mL. Image, protocol and testing courtesy of YCharOS Inc. (ycharos.com).